The application of motivational theory to cardiovascular risk reduction. The level of motivation sustained by an individual has been identified as a primary predictor of success in sustained cardiovascular risk factor modification efforts. This article reviews the primary motivational theories that have been used to explain and predict cardiovascular risk reduction. Specifically, the application of the Health Belief Model, Health Promotion Model, Theory of Reasoned Action, Theory of Planned Behavior and Self-efficacy Theory to the initiation and maintenance of cardiovascular health behavior is addressed. The implication of these theories for the development of nursing interventions as well as new directions for nursing research and practice in the study of individual motivation in health behavior change are discussed.


P-aminodiphenylamine induced biochemical changes in sex organs of male albino rats. Intraperitoneal administration of p-aminodiphenylamine (p-ADPA), an aromatic amine of wide industrial applications, / 42.5 mg/kg body weight for 180 days significantly decreased the activities of testicular lactate dehydrogenase and hyaluronidase and lactic acid content indicating arrest of spermatogenesis. Patchy necrosis of the testis was confirmed histopathologically. No change in testicular cholesterol, fructose content of coagulating glands and dorso-lateral prostate and activities of alkaline phosphatase in seminal vesicle and acid phosphatase in ventral prostate support normal androgenic status.


Electrokinetic energy conversion studies of urine-oxalic acid systems across urinary bladder membrane. Electrokinetic studies of urine-oxalic acid systems with increasing concentration of oxalic acid in urine have been carried out across urinary bladder membranes. It has been found that electro-osmotic flux and streaming current decrease with increase in concentration of oxalic acid in urine while hydrodynamic flux and streaming potential increase with increase in concentration. Kinetic energy term (alpha 1) and polarizability term (alpha 2) have been computed for these systems and it has been found that polarizability decreases much faster with increase in concentration of oxalic acid in urine. Electrokinetic energy conversion of these systems have been computed and it has been found that electrokinetic energy conversion is maximum for urine and it decreases with increase in concentration of oxalic acid in urine. Poor energy conversion may lead to sluggish flushing action which may ultimately lead to formation of urinary calculi in the bladder and so present study may be of some use in predicting electrophysiology of the bladder.


Uranyl nitrate induced corpuscular derangement: an early indication of induced acute renal failure. Erythrocyte structure was studied in rat after uranyl nitrate (UN:5 mg/kg) intoxication. The study of pathogenic progression of UN induced renal failure (ARF) was confined to the early initiation phase (2 hr), late initiation phase (8 hr) and the maintenance phase (24 hr). Erythrocyte structure has been found to be greatly influenced. The UN induced hemolytic syndrome/hypoxia was accompanied by a marked anisocytosis and poikilocytosis during different phases of ARF, which is characteristic of UN poisoning. Subsequent alterations in erythrocyte structure followed by UN administration or during the pathogenic progression of ARF has clinical and diagnostic importance as the alterations were much distinct prior to the clinical manifestation of ARF even at light microscopic level.


Changes in levels of lipid peroxides and activity of superoxide dismutase and catalase in diabetes associated with myocardial infarction. Studies were carried out on the metabolism of lipid peroxides and antioxidative enzymes during diabetes and diabetes superimposed with myocardial infarction. Diabetes was induced using alloxan and myocardial infarction was induced by isoproterenol. In the case of diabetic animals there was a decrease in the levels of lipid peroxides in the heart while in the case of diabetes associated with myocardial infarction it was slightly elevated. The activity of superoxide dismutase and catalase showed a decrease in both the groups. Glutathione showed a fall in the case of diabetes and diabetes associated with myocardial infarction while taurine in heart and ceruloplasmin in the serum was elevated. Histopathological changes in the heart tissue showed some focal changes in the case of both diabetes and diabetes associated with myocardial infarction, but the degree of necrosis was much less than in the case of myocardial infarction.


Hypolipidemic activity of Achyranthus aspera Linn in normal and triton induced hyperlipemic rats. The alcoholic extract of A. aspera, at 100 mg/kg dose lowered serum cholesterol (TC), phospholipid (PL). triglyceride (TG) and total lipids (TL) levels by 60, 51, 33 and 53% respectively in triton induced hyperlipidemic rats. The chronic administration of this drug at the same doses to normal rats for 30 days, lowered serum TC, PL, TG and TL by 56, 62, 68 and 67% respectively followed by significant reduction in the levels of hepatic lipids. The faecal excretion of cholic acid and deoxycholic acid increased by 24 and 40% respectively under the action of this drug. The possible mechanism of action of cholesterol lowering activity of A. aspera may be due to rapid excretion of bile acids causing low absorption of cholesterol.


Antimicrobial activities of mycotoxin MT81 and its structural derivatives. Antimicrobial activities of mycotoxin MT81 and its acetylated, benzoylated and acetic acid derivatives were determined by zone inhibition method. MT81, acetylated MT81 and acetic acid MT81 exhibited sensitivity (31.25, 62.50, 62.50 micrograms/ml respectively) to all the test bacteria. Benzoylated-MT81 did not exhibit any activity even up to 1000 mcg/ml. MT81, its acetylated and acetic acid derivatives exhibited antifungal activity up to 62.5 micrograms/ml and 125.0 micrograms/ml respectively against test yeasts and other fungi. Benzoylated MT81 also exhibited antifungal activity but the degree was comparatively less.


Indole 3-acetic acid production by ectomycorrhizal fungi. Ability of 8 ectomycorrhizal fungi to synthesise indole 3-acetic acid from L-tryptophan and their growth rate were studied. Differences in the levels of IAA synthesis and biomass production among the 8 mycorrhizal fungi were observed. A positive correlation was recorded between IAA level and mycelial growth. The synthesis of IAA and mycelial biomass were maximum on 30th day after incubation. Pisolithus tinctorius and Laccaria laccata exhibited higher amounts of IAA production than other fungi, whereas Amanita muscaria and Rhizopogon luteolus showed least quantity of IAA.


Cholesterol and lipid peroxidation in 3 beta-(2-diethylaminoethoxy)- androst-5-en-17-oneHCl (U18666A) induced cataractogenesis in rats. Decrease in cholesterol was observed in precataractous, cataractous, advance nuclear cataractous and non-cataractous lenses when 3 beta-(2-diethylaminoethoxy)-androst-5-en-17- oneHCl (U18666A) was injected, sc, to rats. Significant increase in lipid peroxidation was observed before the onset of any apparent lenticular opacity in U18666A treated rats. The results suggest that decrease in cholesterol is capable of altering the structural integrity of lens fibers. However, 12.5% decrease in cholesterol and 5% increase in lipid peroxidation observed in non-cataractous lenses indicated that these changes are not sufficient for any apparent opacification.


Effect of antimicrobials on cholesterol synthesis and content in liver and small intestines. The antimicrobials tetracycline, ampicillin and bactrim (cotrimoxazole) decreased HMG CoA reductase activity in liver and small intestines of albino rats. Diminished incorporation of 1, 2, 14C acetate into cholesterol of small intestines in bactrim group was noted. There was a significant fall in cholesterol content of liver, duodenum, jejunum and ileum of the bactrim group and jejunum only in tetracycline group.


Thyroid in intermediary metabolism of the migratory redheaded bunting, Emberiza bruniceps (Brandt). Effect of thyroidectomy and replacement therapy with L-T4, on liver and plasma biochemical constituents of E. bruniceps, was studied during January (recovery phase). Thyroidectomy elevated significantly the levels of plasma glucose, protein, cholesterol, diglyceride, hepatic cholesterol and depressed significantly hepatic free fatty acid without affecting liver and body weights. Treatment of thyroidectomized birds with L-T4 restored liver and plasma constituents, but had significantly depressed plasma phospholipid. These findings suggest that thyroid hormones are critically involved in lipo-regulatory mechanism(s) in E. bruniceps.


Circadian features of carbohydrate metabolism in domestic fowls exposed to weekly 120 degree-shifts of synchronizer schedule. Effects of weekly 8 hr advance- or delay-shifts on the circadian rhythm of plasma glucose, liver glycogen and muscle glycogen in male domestic fowls, beginning at about 3 days of age, were examined. Circadian rhythm in the aforesaid indices of carbohydrate metabolism in control birds was also studied. Blood and tissue samples were collected from birds in all the three groups at 4 hr intervals over a single 24 hr time scale both at 6th and 12th week of age. Plasma glucose and glycogen content in the tissues were determined by employing standard techniques. Cosinor rhythmometry was used for analyzing time series data. In general, a statistically significant circadian rhythm was documented for all the three indices in control and advance-schedule birds, irrespective of age. In contrast, in delay-schedule birds, statistically significant circadian rhythm could not be detected, excluding in muscle glycogen at 12th week of age. The poor growth rate in the delay-schedule birds could be imputed to the disappearance of circadian rhythm in the indices of carbohydrate metabolism.


Ontogenetic and comparative analysis of LDH isozymes in the three species of Indian major carp. Expression patterns of lactate dehydrogenase (LDH) isozyme were investigated in embryonic and post embryonic stages of Labeo rohita, Cirrhinus mrigala and Catla catla using starch gel electrophoresis. Species specific and differential enzyme locus (gene) expression patterns were found in LDH up to 18th hr of study. The isozyme up to 36th hr after fertilization seemed to be completely active and showed electrophoretic patterns very similar to those of the adults. Comparative analysis of the isozyme of the three species permitted species identification even during the embryonic stages when it is impossible to identify on morphological characters only. Also, the genetic studies indicated different taxonomical and evolutionary histories of the species.


Isolation and characterization of an analogue-resistant aminoacyl-tRNA synthetase mutant in Aspergillus nidulans. Six mutants resistant to p-fluorophenylalanine (FPA) were selected on a medium containing aspartate as the sole source of nitrogen using a phenylalanine-requiring (phenA)auxotroph of A. nidulans as the wild type. The mutants, on the basis of genetic characterization, were found to be alleilic and located on the left arm of the linkage group III, approximately 13 map unit left to meth H locus, henceforth assigned to the symbol fpaV. At a fixed concentration of phenylalanine (23 micrograms/ml), the LD50 value of FPA for all the six mutants was found to be about three times more than that for the wild type strain. Affinity chromatographic purification of the enzyme phenylalanyl-tRNA (Phe-tRNA) synthetase from the mutant as well as the wild type strains, revealed that the wild type enzyme had about 1.4-fold higher affinity for phenylalanine as compared to that for FPA, both in the affinity column and in the catalytic reaction. However, the mutant enzyme showed almost a similar affinity for both in columns but a greatly reduced affinity for FPA in the catalytic reaction.


Inactivation of a beta-glucosidase from Arthrobotrys conoides by diethyl pyrocarbonate: evidence of histidine at the active site. Modification of A. conoides beta-glucosidase by diethylpyrocarbonate caused rapid inactivation of the enzyme. The kinetic analyses showed that the inactivation by diethylpyrocarbonate resulted from the modification of an average of one histidine residue per mole of enzyme. The modified enzyme showed an increase in absorbance at 240 nm. Sulphydryl, lysine and tyrosine residues were not modified by diethylpyrocarbonate treatment. The substrate offered significant protection against diethylpyrocarbonates modification. The results indicate that diethylpyrocarbonate was interacting with the enzyme at or near the active site.


Comparison of single breath and steady state methods for the measurement of pulmonary diffusing capacity for carbon monoxide in normal subjects, patients with bronchial asthma and chronic obstructive airway disease. The results of pulmonary diffusing capacity (DLCO) measurements by the steady state (DLCOss) and single breath (DLCOsb) methods were compared in 50 normal subjects (Group A), 50 patients with bronchial asthma (Group B) and 50 patients with chronic obstructive lung disease (COAD) (Group C). A significant correlation was observed between DLCOss and DLCOsb in all the three groups of subjects. This relationship was stronger in Groups A and B as compared to Group C. Both DLCOsb and DLCOss were significantly correlated to parameters of air-flow obstruction. Both measurements of DLCO appear valid in normal subjects and in patients with bronchial asthma. In patients with COAD, on the other hand, DLCOsb may be valid measurement, since it is less likely to be influenced by airways obstruction.


Role of corticosteroids in the management of chronic obstructive lung disease: factors predicting response. A double-blind crossover trial of prednisolone was conducted in 70 patients of chronic obstructive lung disease (COLD). Emphysema dominated the clinical picture in 38, rest being chronic bronchitis with varying degrees of air trapping. None of the patients had clinical asthma. All the patients had obtained maximal benefit from an optimal dose of bronchodilators prior to entering the study. Prednisolone in a dose of 0.8 mg/kg was prescribed in a double blind crossover manner with identical appearing placebo tablets. Patients were evaluated on a weekly basis for an objective as well as a subjective response and side effects of therapy. Thirty-four patients demonstrated a statistically significant improvement in pulmonary functions. In 20 others only subjective response was observed. A good objective response was predicted by a pre-study variability in FEV1, disease duration of less than 10 years and a history of smoking less than 50 pack years.


A study on the prevalence of chronic bronchitis in workers exposed to smoke and irritant fumes in a railway workshop. In the present study 108 full time workers employed in various sections of a railway workshop and exposed to smoke and irritant fumes, and 45 matched control subjects from local administrative staff were investigated to find prevalence of chronic bronchitis. In all 18 (16.7%) workers and 4 (8.9%) control subjects had clinical evidence of chronic bronchitis. Peak expiratory flow rate less than 300 L/min was observed in 54.6% workers and 2.2% control subjects. A significantly higher prevalence of chronic bronchitis was seen in workers. The advancing age, smoking and duration of exposure had significant influence on prevalence of chronic bronchitis.


Re-expansion pulmonary edema and hypotension. A case with diagnosis of non-Hodgkin's lymphoma and superior vena cava obstruction on chemotherapy, presented with respiratory distress and massive pleural effusion of right hemithorax. On removal of 3.5 litres of fluid, he developed pulmonary edema of the same side and hypotension.


Pneumoconiosis in an Indian slate-pencil factory worker. A male slate-pencil factory worker who inhaled dust for 10 years came to our hospital with complaints of breathlessness of two months duration. He developed massive fibrotic lesions in both lungs but was not diagnosed earlier due to the absence of symptoms.


Airway obstruction in bronchiectasis and its reversibility--a study of 38 patients. Thirty-eight patients (20 males and 18 females) with bronchiectasis were studied for the evidence of airway obstruction and its reversibility in response to subcutaneous terbutaline. There was an improvement in VC (8.56%), FEV1 (6.36%), PEFR (15%) and PEF 25-75% (18%) respectively. The difference being statistically significant in VC and highly significant in FEV1, PEFR and FEF25-75 as compared to initial values and in 6 patients the improvement in FEV1 was more than 15%. The improvement in airway obstruction was not related to the character of expectoration. It is suggested that all the patients of bronchiectasis should be assessed for the reversibility of airway obstruction and those showing a reversible pattern be treated accordingly.


A seroepidemiologic study of hepatitis A in Spanish children. Relationship of prevalence to age and socio-environmental factors. Three child population groups from the Madrid area were studied for anti-HAV antibodies. Analysis was carried out with respect to age and socio-environmental factors. The population understudy was composed of 156 children, with ages ranging from 1 to 14 years; they were stratified in three socio-environmental groups (white-family unit, gypsy-family unit and orphanage), and also divided into subgroups according to age. As a whole, an age-related increase in prevalence was found. The overall seroprevalence by socio-environmental groups was: gypsy-family unit 63%, orphanage 46%, and white-family unit 23%. Significant differences between groups appeared from seven years on, being more marked among the eldest subgroups. Among the factors evaluated, hygienic-sanitary conditions and overcrowding influenced the high prevalence rate found in the gypsy-family unit subjects, whereas overcrowding appeared to be responsible for the higher prevalence in orphanage residents, as compared to white-family unit children.


Borrelia burgdorferi infection in Europe: an HLA-related disease? Various studies in the United States and Europe have established human leucocyte antigens (HLA) Cw3, DR2 and DR4 as risk factors for manifest Borrelia burgdorferi infection or the development of chronic courses of Lyme disease. Other studies failed to confirm these findings. In the present study the frequencies of HLA A, B, Cw and DR were analysed in 283 persons from Austria and Germany with manifest B. burgdorferi infection. No statistically significant differences were found between patients and the control groups with regard to the frequencies of particular HLA antigens, nor were differences in antigen frequencies in the patients with manifestations of different stages of disease significant. Furthermore, a statistical re-evaluation of all the European studies failed to confirm particular HLA antigens as risk factors for B. burgdorferi infections to become manifest or chronic.


Solitary borrelial lymphocytoma: report of 36 cases. Thirty-six cases of borrelial lymphocytoma were detected during the period 1986 to 1990 in Slovenia. Borrelial lymphocytoma was located on the ear lobe in 17 persons, ten female and seven male, with a median age of 12 years (range 2-56). Fourteen of these 17 were children under 14 years of age. A tick bite was remembered by 15 patients a median of 30 days before borrelial lymphocytoma developed. The most frequent month of onset was September. Erythema migrans preceded or accompanied borrelial lymphocytoma in eight cases. In 15 cases, eight female and seven male, borrelial lymphocytoma was localized on the mamilla. Median age of these patients was 42 years (range 15-72). Twelve had a tick bite about 45 days (median value) before the onset of borrelial lymphocytoma, which occurred most frequently in August. Erythema migrans was reported in 13 patients and preceded borrelial lymphocytoma in ten cases. In another four patients borrelial lymphocytoma was localized on the nose, scrotum, upper arm and shoulder. Antibiotic treatment with phenoxymethyl-penicillin (n = 16), ceftriaxone (n = 8), doxycycline (n = 9), azithromycin (n = 2) and penicillin G (1) led to complete recovery within an average of three weeks in all cases.


Acyclovir monotherapy versus acyclovir plus beta-interferon in focal viral encephalitis in children. Severe focal viral encephalitis is most commonly caused by herpes simplex virus (HSV), but other viruses may act as etiologic agents as well. Acyclovir (ACV) is the standard therapy for HSV encephalitis, but the mortality of 28% and defect healing rate of about 35% are still unsatisfactory. Furthermore, ACV has virtually no effect on other pathogens of viral encephalitis, except for varicella-zoster virus (VZV). It is well known that beta-interferon (beta-IFN) has a broad antiviral spectrum, and it has been demonstrated in vitro that beta-IFN in combination with acyclovir has synergistic inhibitory effects on HSV. To investigate if the combination of ACV with and without beta-IFN might also be of significance for the treatment of severe viral encephalitis, we performed a retrospective study. A case record form was sent to all 278 West German children's hospitals. The response rate was 78%. A total of 301 patients were reported, of whom 214 received specific antiviral therapy with either ACV alone (n = 179) or ACV plus beta-IFN (n = 35). No overall differences between ACV monotherapy and the combination therapy were observed. However, in a subgroup of 41 patients (ACV n = 30, ACV plus beta-IFN n = 11) who had low-density areas of the temporal lobes on cranial computed tomography scans, compatible with severe focal encephalitis, sequelae due to defect formation and mortality were significantly (p = 0.014) reduced in patients who had received combination therapy.(ABSTRACT TRUNCATED AT 250 WORDS)


Bacteremia caused by Stomatococcus mucilaginosus: report of seven cases and review of the literature. During a three-year period eight patients with blood cultures positive for Stomatococcus mucilaginosus were identified at two university hospitals. One patient without any signs of infection had a central venous catheter that was colonized with this organism, two patients had transient bacteremia without definite relationship to underlying disease, whereas the remaining five patients suffered from clinically significant infections. Of these last five patients, one had undergone prior head and neck surgery and four had hematologic malignancy with mild to severe neutropenia; two of the latter patients developed the infection subsequent to dental surgery. Besides neutropenia and mucosal damage in the oropharynx, quinolone antibacterial prophylaxis may have been an additional risk factor for the development of S. mucilaginosus bacteremia in these patients. A thorough review of the literature revealed that in addition to our findings, endocarditis and foreign body infections are further typical clinical manifestations. Although the overall antibiotic susceptibility pattern of S. mucilaginosus resembles that of streptococci, it is suggested that penicillin G may not be the drug of choice for initial therapy of particularly severe infections. S. mucilaginosus can be easily differentiated from other gram-positive bacteria when certain key criteria (e.g. adherence to agar surfaces, poor growth on Mueller-Hinton agar, presence of a capsule) as well as an array of biochemical tests, including commercially available identification systems, are applied. Our own and published data emphasize that both microbiologists and clinicians should be increasingly aware of this opportunistic pathogen.


The antimicrobial spectrum of honey and its clinical significance. The antimicrobial spectrum of honey was investigated by placing two drops into each of the wells made on culture media on which pure cultures of various organisms obtained from surgical specimens were grown. The organisms were grown under both aerobic and anaerobic environments. Fungal cultures of common fungi causing surgical infections or wound contaminations were mixed with 100%, 50% and 20% unprocessed honey. Growth inhibition was complete in the media containing 100%, partial in media containing 50% and no inhibition was produced by 20% honey. Unprocessed honey inhibited most of the fungi and bacteria causing wound infection and surgical infection except Pseudomonas aeruginosa and Clostridium oedematiens. Apart from Streptococcus pyogenes which is only moderately inhibited, golden syrup, a sugar syrup with similar physical properties as honey, did not inhibit any of the bacteria or fungi tested, demonstrating that honey is superior to any hypertonic sugar solution in antimicrobial activity. Honey is thus an ideal topical wound dressing agent in surgical infections, burns and wound infections.


Temporofacial zygomycosis in a pregnant woman. A 38-year-old Omani woman, seven months pregnant, developed extensive zygomycosis involving maxillary and temporal bones. No evidence of any underlying immune deficiency was detected except that which may be attributed to pregnancy. After delivery the patient was treated with repeated courses of amphotericin B, which resulted in a complete clinical resolution. Zygomycosis in uncomplicated pregnancy has not been previously described.


Prevalence of HCV antibodies in health-care workers from northern Italy. The prevalence of HCV antibodies was assessed in 407 health-care workers and in 253 control subjects by means of immunoassays based on recombinant antigens. The seroprevalence in the study group was fairly low (1.2%) and not statistically different from that of controls (0.8%). The relation of HBV and HCV infections was evaluated in 83 health-care workers and in 82 controls: in both groups anti-HCV positivity was weakly related (p less than 0.05) to the HBV infection. HCV infection was associated with working in high risk wards (4/5 cases) and with report of accidental needle pricks (4/5 cases). As evaluated by means of the available markers, HCV infections in health-care workers seem to be rare.


Massive renal infarction due to mucormycosis in an AIDS patient. Isolated renal infection by Mucor spp. occurs very rarely, even in the immunocompromised host. The mortality of the different forms of mucormycosis reaches 75-100% in most series. Infection of the kidney is usually diagnosed post-mortem. Thus, when renal infection is recognized, aggressive treatment including surgery and amphotericin B is required. We present a case in which a massive left renal infarction was the unique manifestation of mucormycosis in an AIDS patient. Administration of amphotericin B and left nephrectomy were necessary to achieve satisfactory outcome.


Effect of dialyser composition and reuse on neutrophil count and elastase alpha-1 proteinase inhibitor complex formation. To assess the inter-relationship of leucopenia and PMN elastase release we undertook a prospective crossover study of 6 patients dialysed with new and reused cuprophane, cellulose acetate and polysulfone membranes. Serial blood samples were analysed for PMN count, and elastase-alpha 1-proteinase inhibitor complex (E alpha 1PI) concentrations. After 15 min dialysis with new membranes median PMN counts fell by 72.2%, 25.3% and 22.1% with cuprophane, cellulose and polysulfone, respectively. With reuse the decreases were reduced to 6.4%, 8% and 13.6%. All membranes produced a gradual increase of E alpha 1PI. Median E alpha 1PI accumulation rates (ng ml-1 min-1) with new membranes were 175, 169 and 187 for cuprophane, cellulose acetate and polysulfone, respectively. With reuse of cuphrophane and cellulose acetate these rates fell to 99 and 109 (p less than 0.05 and p less than 0.05, respectively), however, with polysulfone it remained unchanged at 180 ng ml-1 min-1. This study highlights differences between two aspects of the neutrophil response to haemodialysis, and demonstrates that extrapolation from individual parameters to conclusions concerning biocompatibility may be inappropriate.


Failure of blood pressure to increase following erythropoietin therapy in the renoprival status. Two anephric patients in the course of one year erythropoietin therapy improved their anemic status without changes in Mean Arterial Blood Pressure. The discordant time course behaviour of hematocrit and blood pressure points to the importance of residual renal tissue for blood pressure to develop during erythropoietin therapy in the renoprival status.


Mechanical bridge to transplantation with the Vienna heart in TAH and LVAD configuration. The Vienna heart uses a vacuum formed, pellethane pulsatile ventricle and is available in left ventricular assist (LVAD) and total artificial heart (TAH) configurations. This device was used as mechanical support of the failing heart in nine patients intended for heart transplantation. In two patients with cardiomyopathy an orthotopic TAH was implanted; one survived despite severe preoperative ischemic liver damage, and the other died of sepsis. In seven patients an atrio-aortic LVAD was implanted; six had suffered an acute myocardial infarction with cardiogenic shock, and one could not be weaned off bypass. Three patients survived. These included one 65-year-old with incipient ARDS at operation, and a 40-year-old with preoperative liver and kidney insufficiency who was transplanted in septicemia. In this patient the septic focus, natural and artificial heart, were removed at transplantation. Four patients died. In one we were unable to establish satisfactory circulation, one died after failure of the transplanted heart, one suffered a lethal cerebral embolism and one developed multi-organ failure after repeated attacks of ventricular fibrillation. With the Vienna heart sufficient circulatory support could be established with cardiac outputs between 6 and 8 l/min for the TAH and 3.5 to 4.5 l/min for the LVAD. With this type of support an overall survival rate of 44% could be achieved. Mechanical hemolysis was not a clinical problem and no device failure occurred.


Clinical effects of a sorbent suspension dialysis system in treatment of hepatic coma (the BioLogic-DT). Fifteen patients with acute deterioration of liver function, high serum ammonium, and an average coma level of 3.9 were identified. Eleven of the patients were on respirator support, and eleven had kidney failure pursuant to the liver failure. The patients were treated for 8-12 hours daily with the BioLogic-DT system, in which membranes of a cellulosic plate dialyzer actively pump blood through a single access at over 200 ml/min, and the dialysate contains a suspension of powdered activated charcoal (300,000 square meters surface area) and cation exchanger (160 meq capacity). No anticoagulant was used. In spite of the declining condition of the patients prior to treatment, there was a statistically significant improvement in neurologic status during individual treatments, and a positive trend over 1-12 (average four) daily treatments. Four patients recovered liver function and another four improved enough to receive a liver transplant operation. The BioLogic-DT system appears to be safe in treatment of patients with hepatic insufficiency and coma. The neurologic improvement of these patients indicates that many toxins of hepatic failure are dialyzable across cellulosic membranes and bound by charcoal.


Microcarrier culture of hepatocytes in whole plasma for use in liver support bioreactors. Contact activation of plasma clotting may limit the use of some microcarrier types for hepatocyte attachment in a liver assist device. Activation by seven microcarrier types was studied in plasma containing 2-500 units heparin/mL. Clotting was activated by dextran (Cytodex 1 and 2) and collagen-coated (Cytodex 3) microcarriers at 2-25 units/mL (Cytodex 1) and 2-100 units/mL (Cytodex 2 and 3). There was no activation by polystyrene, gelatin, glass or fibronectin-coated polystyrene microcarriers. Compared with culture medium, incubation of HepG2 cells in plasma did not affect cell viability but increased cell number (56.4 versus 65.1 x 10(4) cells; P less than 0.05) and incorporation of [3H]-amino acids into protein (204913 versus 279624 dpm; P less than 0.05). Polystyrene-attached cells demonstrated time-linear protein synthesis, glucose and 7-ethoxycoumarin metabolism. We conclude that polystyrene-attached hepatocytes maintain viability and metabolic activity in plasma and are of potential use in a liver support bioreactor.


Spontaneous dialytic ultrafiltration with intraperitoneal reinfusion of the concentrate in 15 cirrhotic patients with intractable ascites. The clinical efficacy and tolerance of dialytic ultrafiltration of ascites through a hemofilter (DUF) with peritoneal reinfusion of the concentrate was evaluated in 15 cirrhotic patients with intractable ascites. All together, 51 DUF procedures were carried out. An average of 8.6 was ultrafiltered during 12 h with no significant change in blood pressure, hemoglobin, coagulation parameters or plasma creatinine. A significant increase in ascitic protein concentration was observed immediately after the procedure and a slight but significant increase in 24 h urinary output. A controlled evaluation of DUF compared to large paracenteses seems to be justified by these preliminary results.


Plasma exchange treatment in rapidly progressive glomerulonephritis associated with anti-neutrophil cytoplasmic autoantibodies. This study reports on 12 patients with acute renal failure due to biopsy-proven rapidly progressive glomerulonephritis and signs of systemic disease in whom antineutrophil cytoplasmic autoantibodies (ANCA) were detected by indirect immunofluorescence (IIF) on alcohol-fixed neutrophils and assessed in serial determinations by ELISA. The diagnosis was: Wegener's granulomatosis in nine patients who showed a diffuse cytoplasmic pattern at IIF (c-ANCA), and microscopic polyarteritis in three where a perinuclear pattern (p-ANCA) was seen. All patients underwent a course of plasma exchange - PE - (3-10 sessions per patient) associated with steroids and cyclophosphamide. The ANCA titer dropped steeply during PE in all cases and was followed by disappearance of systemic symptoms and renal function improvement within four weeks. After a follow-up period of 50 +/- 31.2 months all patients were alive without signs of disease activity; ten had stable renal function, with serum creatinine 1.8 +/- 0.7 mg/dl; two had entered regular dialysis treatment after 44 and 82 months. Our results suggest that the rapid removal of ANCA by means of PE can help control disease activity and reduce the risk of death or end-stage renal disease.


Changes in coagulation factors by passage through a dextran sulfate cellulose column during low-density lipoprotein apheresis. To determine the extent of adsorption of coagulation factors by a dextran sulfate cellulose column used for selective removal of low-density lipoprotein (LDL), various coagulation factors were measured before and after application to the column during LDL apheresis. The column almost completely adsorbed many coagulation factors. Although the bradykinin concentration was markedly increased by passing the plasma through the column, this increment was suppressed by nafamostat mesilate which inhibits the initial contact phase of the intrinsic coagulation pathway. The von Willebrand factor, which forms a complex with factor VIII in plasma, is reduced in apheresis with nafamostat mesilate to the same extent as in apheresis without nafamostat mesilate. Thus, coagulation factors seem to be adsorbed by different mechanisms which include activation of the initial contact phase by the negative charges of dextran sulfate and concomitant adsorption with the phospholipid portion of lipoproteins containing apolipoprotein B or with von Willebrand factor.


Novel quantitative methods for the determination of biomaterial cytotoxicity. Two novel methods for the determination of biomaterial cytotoxicity using cell culture are presented. The methods combine a standardized protocol for producing extracts from medical devices with either the established MTT assay or a new fluorimetric assay. The suitability of both methods for evaluating the toxicity of candidate materials was demonstrated by resolution of the differences in the toxic effects of serial dilutions of a PVC extract on BHK21 and HT1080 cells. The tests yield highly reproducible, quantitative results and can be applied to materials in the usual physical forms applicable to artificial organs.


p53 over-expression is an early event in the development of human squamous-cell carcinoma of the larynx: genetic and prognostic implications. The tumor-suppressor protein p53 is over-expressed in a large fraction of squamous-cell carcinomas of the larynx (LSCCs). p53 overexpression is dependent upon the synthesis of mutated versions of the protein and has been associated with the malignant progression of certain tumor types. In order to examine the prognostic value of p53 immunodetection in LSCCs, we performed a retrospective analysis on a selected series of tumors, using the PAb 1801 and CM1 antibodies. No significant difference in the frequency of p53 over-expression was observed between tumors from patients with early relapse (67%) and those who had been disease-free for more than 5 years (84%). The lack of correlation of p53 immunoreactivity with clinical stage and differentiation grade of LSCCs, together with the coordinated expression of p53 in primary tumors and the corresponding lymph-node metastases, indicate that p53 over-expression is probably unrelated to the biological aggressiveness of these tumors. In addition, the detection of p53 immunostaining in pre-invasive areas as well as in preneoplastic lesions suggests that p53 abnormalities probably constitute a very early event in LSCC development.


pS2 protein and steroid hormone receptors in invasive breast carcinomas. We have used immunohistochemical and hormone-binding techniques to determine the presence of estrogen receptors, estrogen-receptor protein, progesterone receptors and pS2 protein in 97 invasive breast cancers. Our group of tumors displayed the same frequency of estrogen receptors and progesterone receptors as other comparable groups, but it contained more tumors containing pS2 protein. We also observed staining of morphologically normal cells in lobules adjacent to the tumors and in several fibroadenomas; both findings vary from some other reports. We attribute these variations to the use of a sensitive immunohistochemical method and choice of the lowest possible threshold to classify a tumor as pS2-positive. If we used a higher threshold, then about 95% of the tumors containing pS2 protein also contained estrogen receptor protein. Our results add further weight to the assertion that tumors containing pS2 protein also display estrogen receptors. The data also provide theoretical and indirect support to the clinical prediction that tumors containing pS2 protein are more likely to respond to hormonal therapy and may have a more indolent course than tumors lacking the molecule.


The study of markers of biological effect in cancer prevention research trials. Biological markers may provide a valuable tool for the development of cancer prevention agents, for monitoring patient compliance to a selected intervention, or for further defining the carcinogenic process. This discussion focuses on markers of biological effect and the rationale for their use in cancer prevention trials. Recent studies with biological markers are investigating their incorporation into phase-I, -II, and -III chemoprevention clinical trial designs. Their use in clinical studies is expected to increase the number of agents that may be evaluated and to provide valuable information on the biological effectiveness of agents, doses, and schedules. Markers may also provide information to help in selecting high-risk groups for prevention research, and to indicate the pathways inhibited and the stage of carcinogenesis affected. Such information may prove of crucial importance in strengthening the rationale for long-term trials and other ancillary research. Biomarker research for colon carcinogenesis is discussed, including examples of a number of recent trials that may influence future progress in this area of prevention research. A crucial step in this process is marker validation as an aspect of major prospective observational and intervention studies where cancer incidence is the endpoint. We cannot be fully confident of markers as intermediate endpoints until the evidence from clinical trials is sufficiently strong to support major public health initiatives for prevention.


Phenotypic overlaps between pleomorphic malignant T-cell lymphomas and mixed-cellularity Hodgkin's disease. Histologically diagnosed, or in part questionable, malignant pleomorphic peripheral T-cell lymphomas (pPTCLs, n = 16) and mixed-cellularity Hodgkin's disease (MCHD, n = 12) were objectively compared by the use of combined immunohistochemistry on paraffin sections, test-point analysis of tissue components, and semi-automated nuclear morphometry on semi-thin resin sections. Classical, qualitative histomorphological distinction of these sub-types of lymphomas proved to be valid and is probably still the best method. Quantitative discriminant features, in order of decreasing significance, were: (i) expression by large atypical cells (LACs) of CD45R0, CD43 and CD45 in pPTCLs, and of CD30 and CD15 in MCHD; (ii) means and standard deviations (SDs) of LAC nuclear-profile areas (greater in MCHD than in pPTCLs); (iii) expression of CD3 by LACs in pPTCLs; (iv) prominence of small lymphoid cells in MCHD; (v) higher percentage of medium-sized lymphoid cells in pPTCLs; and (vi) higher SDs of nuclear-profile circularity factor of small lymphoid cells in MCHD. The medians of the largest nucleolar profile areas in LACs per field did not differ in pPTCLs and MCHD, but dispersion of individual values towards higher levels was significantly greater in the latter. Stepwise discriminant analysis of test point and nucleometric variables that best distinguished pPTCLs from MCHD revealed considerable overlaps, and questionable cases tended to be intermediate between the two. In conclusion, our results confirm and expand the notion of intra-group heterogeneity, with indistinct borders and the existence of intermediate phenotypes between these two taxonomic categories of malignant lymphomas.


Determination of placental ferritin (PLF)-positive lymphocytes in women in early stages of breast cancer. Previous studies have proved that a certain acidic isoform of ferritin is specifically synthesized by the placenta and breast-cancer tissue. In this context it has been further reported that the determination of this so-called placental isoferritin (PLF) on the surface of a subset of peripheral lymphocytes is highly specific and sensitive for early stage breast cancer. By use of the monoclonal antibody CM-H-9 and flow cytometry, the levels of placental ferritin (PLF)-positive cells were determined in 133 female patients undergoing surgical excision of a controversial or highly suspicious lesion of the breast detected by mammography. In addition, 61 healthy blood donors served as controls. Values of PLF-positive cells in breast cancer patients differed significantly from those found in women with benign diseases and healthy controls (3.87% vs. 1.55% and 2.02, respectively; p less than 0.00001). Analysis of prognostic factors in breast cancer patients (tumor size, lymph-node status, menopausal status, estrogen receptor status, histologic grading and grading subfactors) revealed significantly higher levels of PLF-positive cells in lymph-node-negative patients compared with node-positive patients (p less than 0.00001). Furthermore, levels of PLF-positive cells showed a significantly negative correlation with tumor size and nuclear polymorphism. In 15 patients who underwent a guide-wire-directed surgical biopsy for a non-palpable, mammographically suspect lesion, 4 cases of cancer correlated with high values of PLF-positive lymphocytes while only 1 patient with a benign histologic result exhibited more than 4% positive cells.


Matrigel promotes retinoblastoma cell growth in vitro and in vivo. Cells derived from retinoblastomas grow slowly in vitro and only very rarely form tumors in nude mice. Matrigel, a mixture of components normally found in basement membranes, promotes the growth of Y-79 and WERI-Rb1 retinoblastoma (Rb) cells when added to suspension cultures of the 2 Rb cell lines. It also substantially increases cell adhesion in vitro. Y-79 cells, seeded into a Matrigel matrix, form round colonies over a 3-week period similar to those of control, weakly metastatic murine melanoma cells. In vivo, s.c. co-injection of Matrigel with either Y-79 or WERI-Rb 1 cells into nude mice promotes retinoblastoma tumor formation. Transplantation of as few as 1,000 cells allows for xenografting under these conditions, while no tumors were observed in the absence of Matrigel, even at 10 x 10(6) cells/inoculum. The tumors produced have the expected morphology and express an mRNA for a highly specific retina/retinoblastoma marker protein, the interphotoreceptor retinoid-binding protein. Thus, the xenografts obtained maintain the original morphological and molecular characteristics of the injected cells and represent a useful model for in vivo studies of retinoblastoma growth and treatment.


Influence of recombinant human interleukin (IL)-7 on disease progression in mice infected with Friend virus complex. Recombinant human (rhu) IL-7 was evaluated for its influence on disease progression in mice infected with the polycythemia-inducing strain of the Friend virus complex (FVC). DBA/2 mice were injected i.v. with FVC, and then treated s.c. with rhuIL-7. IL-7 significantly prolonged survival time and decreased spleen focus-forming virus (SFFV) levels, expression of SFFV mRNA and SFFV protein production in FVC-infected mice. IL-7 did not appear to directly inactivate SFFV. Although both splenic weight and cellularity in FVC-infected mice treated with IL-7 were higher than those of normal mice, they were respectively 58% and 66% lower than those of the untreated FVC-infected mice. NK-cell activity was substantially lower in FVC-infected mice than in normal mice, while IL-7 restored NK-cell activity to normal levels. IL-6 and IFN-gamma levels were markedly reduced in FVC-infected mice compared to normal mice, but treatment of FVC-infected mice with IL-7 restored these cytokine levels. While the actual mechanisms of these effects are not yet known, the results suggest the potential therapeutic efficacy of IL-7 for certain hematopoietic and viral disorders, possibly mediated through an action on accessory cells and cytokine production.


Biodistribution of lymphokine-activated killer (LAK) cells in Wag rats after hepatic-artery or jugular-vein infusion. This study deals with the biodistribution of syngeneic radiolabeled lymphokine-activated killer (LAK) cells in Wag rats after infusion via the hepatic artery or the jugular vein. The biodistribution of 111Indium-labeled LAK cells was evaluated using serial whole-body gamma camera imaging. Furthermore, we investigated 2 factors that might influence the biodistribution of these effector cells: purity of LAK cells and administration of interleukin-2 (IL-2). After injection of 111Indium-labeled LAK cells via the hepatic artery or via the jugular vein we could detect important differences in the biodistribution pattern up to 5 hr after injection. LAK cells administered via the jugular vein were all found in the lungs up to 2 hr after injection and then redistributed to the liver and to the spleen. LAK cells administered via the hepatic artery were all found in the liver after injection and redistributed after 2 hr mainly to the spleen. About 8 hr after injection we could no longer detect any differences in the biodistribution pattern according to the route of administration. Biodistribution was followed for up to 72 hr after injection but the pattern showed no change after 8 hr, whichever the route of administration. A purified adherent-LAK population, a large granular lymphocyte culture with only 6% T cells, showed the same distribution pattern as standard LAK cells (40% T cells). Infusions of 40,000 units of IL-2 per day, starting 3 days before and continuing after administration of radio-labeled LAK cells, accelerated the redistribution of these cells by both routes of administration. We conclude that up to 2 hr after local infusion, a high concentration of LAK cells in the first capillary bed can be obtained. Therefore, local administration of LAK cells may be more effective against tumors.


Influence of glucose on metabolism and growth of rat glioma cells (C6) in multicellular spheroid culture. The metabolism and growth of rat glioma C6 cells in multicellular spheroid culture depended strongly on the glucose supply. A low glucose level (0.1 g/l) in the culture medium reduced lactate production, increased oxygen consumption and diminished hydrogen ion production under normoxia as well as hypoxia. A high glucose level (10 g/l glucose) increased lactate production, had no significant influence on oxygen consumption and increased the hydrogen ion production under hypoxia. Hydrogen ion release from cells under normoxic and hypoxic conditions could be significantly diminished by amiloride (l mM), indicating the involvement of the Na+/H+ exchanger. The growth of the C6 spheroids was enhanced under low glucose conditions, possibly due to the more physiological extracellular pH in the deeper regions of the spheroids. The growth was inhibited under high glucose conditions, which seemed to be toxic due to a massive hydrogen production giving acidosis. The glucose supply strongly influenced the local hydrogen ion production inside the C6 spheroids and this might in turn lead to changes in the response to different therapeutic modalities.


Effects of short-chain fatty acids on growth and differentiation of the human colon-cancer cell line HT29. Short-chain fatty acids (SCFAs), namely butyrate, acetate and propionate, originate from the bacterial fermentation of dietary fibers and are the predominant anions present in the large bowel. Our study was carried out to investigate the effects of SCFAs on growth of the human adenocarcinoma cell line, HT29. The results show that, under our culture conditions, both propionate and butyrate inhibit growth of HT29 cells, whereas acetate has no significant effect. The antiproliferative effect of propionate or butyrate is associated with an inhibition of FCS-induced activation of ornithine decarboxylase (ODC), a key enzyme of polyamine metabolism. Inhibition of growth induced by either propionate or butyrate is not reversed by the addition of putrescine, which reveals that these SCFAs are not acting solely on the ODC/polyamine system. Our data show that propionate and butyrate, unlike acetate, induce an increase in alkaline phosphatase activity, which reflects a more differentiated phenotype than that of untreated control cells. Taken together, our results suggest that propionate, like butyrate, may play an important role in the physiology of the colon and could partially account for the protective effect of dietary fibers with respect to colon carcinogenesis.


Derivation of tumorigenic and non-tumorigenic mouse alveolar type-II cell lines from fetal type-II cells after a combined in vivo/in vitro carcinogen treatment. Alveolar type-II cells were isolated from the lungs of fetuses (day 18 of gestation) of the A/WySnAf (A/Sn) mouse strain, which were treated in utero at day 15 with the directly-acting carcinogen N-ethyl-N-nitrosourea (ENU). The isolated type-II cells were again treated with ENU during their initial growth in vitro. After a prolonged culture period, 5 cell lines were obtained, which were identified as type-II cell lines. Differences between cell lines were found with respect to contact-inhibited growth, cell doubling time and ability to grow in a serum-free medium. Two out of the 5 cell lines produced highly invasive type-II cell carcinomas after s.c. injection of 5 x 10(6) cells into nude mice. Thus, both tumorigenic and non-tumorigenic mouse alveolar type-II cell lines were derived after this combined in vivo and in vitro carcinogen treatment of fetal mouse alveolar type-II cells. This offers the possibility of studying in vitro the factors thought to influence lung tumorigenesis in vivo. In addition, our findings strongly suggest that alveolar type-II cells are the progenitor cells of malignant mouse lung tumors.


Effects of areca nut on growth, differentiation and formation of DNA damage in cultured human buccal epithelial cells. Because the high incidence of oral cancers in South-East Asia is causally linked to the common habit of betel quid chewing, the effects of an aqueous extract of areca nut, one of the main ingredients of the quid, on growth, differentiation, morphology and DNA damage were studied in cultured human buccal epithelial cells. An acute exposure (3 hr) of the cells to the extract altered their morphology and induced ridges in the plasma membrane, with indications of internalization of extract particles. Such exposure also caused formation of DNA single-strand breaks which accumulated during post-treatment culture, indicating continuous exposure to residual particles and/or the possibility of inhibited DNA repair. The extract accelerated terminal differentiation of the cells, measured as involucrin expression at relatively non-toxic levels. The extract caused similar loss of colony-forming efficiency in normal cells and in a buccal carcinoma cell line (SqCC/YI) which was defective in its ability to undergo differentiation, indicating that extract toxicity could occur independently from this response. Finally, the genotoxicity of the salivary areca-nut-specific carcinogen 3-(N-nitrosomethyl-amino)propionaldehyde, was demonstrated by the formation of DNA protein cross-links and DNA single-strand breaks in normal buccal epithelial cells. These findings in vitro suggest that betel quid carcinogenesis in the human oral cavity may involve cytopathic alterations of normal cell morphology, growth and differentiation, as well as formation of DNA damage by areca-nut-related agents extracted or formed in saliva.


Transforming activity and the level of Tax protein: effect of one point mutation in HTLV-I tax gene. The transcriptional trans-activator molecule of HTLV-I, Tax, is known to transform rodent fibroblasts. A revertant clone expressing Tax was obtained by treating transformed Rat-I cells harboring a single copy of the tax gene with a mutagen. Sequence analysis of the tax gene of the revertant clone revealed that it had one point mutation at codon 12(CTT----TTT), resulting in a change from Leu to Phe. The colony-forming efficiencies of the cells transfected with the mutant tax gene (mu71 tax) were significantly lower than those transfected with the wild-type by the soft-agar method. This difference was shown to be due to the instability of mu71 Tax.


The H-ras oncogene regulates expression of 70- and 45-kDa cell-surface molecules whose expression correlates with tumor-cell immunogenicity. The effects of the H-ras oncogene on fibroblast cell tumorigenicity and immunogenicity was studied in transfectants of the BALB/c 3T3 clone A31 fibroblastoid cell-line. Cells that were transfected with MC29-LTR-H-ras (98/6) or MC29-LTR-v-myc + H-ras (98/4v) and were inoculated into syngeneic BALB/c mice were tumorigenic in 100% and 60% of animals respectively. By contrast, transfectants containing the pSV2neo plasmid alone (98/1) displayed normal characteristics both in vitro and in vivo. Inoculation of mice with mitomycin-C-treated 98/1 or 98/4v cells induced an effective protective immunity to a challenge of live 98/4v cells, and a partial immunity against 98/6 cells. Mitomycin-C-treated 98/6 cells failed to render immunity against a challenge of either 98/6 or 98/4v cells. To correlate immunogenicity and tumorigenicity of the different cell types with cell-surface-antigen expression, we prepared MAbs against 98/4v cells in syngeneic mice. Immunohistochemical and immunoblot analysis revealed that MAbs 102 and 104 recognized 2 protein band of 70 and 45 kDa respectively, which were expressed predominantly in 98/1 and 98/4v cells. A third immunoreactive protein band of 44 kDa that reacted with MAb 6 was expressed at a similar cell-surface density on all cell types. Cell-differentiation-inducing agents, such as DMSO, retinoic acid or sodium butyrate, were all found to induce 98/6 cell flattening and morphological changes toward a normal phenotype that were followed by up-regulation of the 70- and 45-kDa antigens. The results suggest that regulation of expression of the 70- and 45-kDa molecules is affected by H-ras, and that expression of these cell-surface molecules may be relevant to tumor cell immunogenicity.


Effects of experimental conditions on the production of interleukin-1 alpha and -1 beta by human endothelial cells cultured in vitro. We have characterized the production of IL-1 alpha and -beta in primary and passaged cultures of quiescent human umbilical vein endothelial cells (HUVECs) using highly specific and sensitive solid-phase enzyme immunoassays. Primary cultures produced both immunoreactive IL-1 alpha and IL-1 beta following stimulation with lipopolysaccharide with the alpha form predominating over the beta. Most of the IL-1 produced remained cell-associated. Primary, but not passaged, cultures were significantly contaminated by macrophage-like cells, possibly accounting for higher production of IL-1, especially IL-1 beta. Gel filtration of secreted proteins derived from cultured HUVECs showed that the immunoreactive IL-1 alpha exhibited the expected molecular weight (17 kDa), but cell-associated IL-1s appeared to be a mixture of the 17 kDa protein and of higher molecular weight precursors. Mitogens in the culture medium (serum and endothelial cell growth supplement) were powerful stimuli of endothelial IL-1 production and accounted for the relatively high basal IL-1 levels observed in the cultured endothelial cells. The proliferative phenotype of the endothelium is possibly linked to the expression of high level of IL-1, which until now was thought to be an autocrine inhibitor of endothelial cell mitosis.


Enhanced induction of lymphokine-activated killer activity after lentinan administration in patients with gastric carcinoma. In 15 patients with gastric carcinoma, peripheral blood mononuclear cells (PBM) were obtained serially before and 3, 5 and 7 days after lentinan administration. The generation of lymphokine-activated killer (LAK) activity, induced by in vitro activation of PBM with interleukin 2 (IL 2), was significantly augmented 5 days after a single intravenous dose of 2 mg lentinan, when compared with that before lentinan injection. Natural killer (NK) activity of PBM was also significantly enhanced 7 days after the drug injection. However, the distribution of lymphocyte subsets exhibited no significant change following lentinan administration.


Aromatase inhibitors regenerate the thymus in aging male rats. The thymus can be regenerated in aging rats by surgical or chemical castration and regeneration is inhibited by testosterone, which may exert this effect, at least in part, through its conversion to estradiol. An attempt has been made to regenerate the thymus in intact aging rats using inhibitors of the aromatase system, in the hope that this maneuver could lead to the use of such chemical intervention in the treatment of immunodeficiency syndromes. Young adult and aging (18-month-old) male rats were orchidectomized under ether anesthesia and 7 days later given s.c. implants of testosterone in silicone elastomer (SILASTIC) tubing. Some rats received testosterone together with a five-fold excess of the aromatase inhibitor 1,4,6-androstatriene-3,17-dione (ATD). One group of young intact rats received implants containing 25 mg ATD and a group of 18-month-old intact rats received 125 mg ATD or 25 mg of another, more powerful aromatase inhibitor 4-hydroxyandrostenedione (4-OH). On the 28th day after implanting, rats were killed and the thymus, spleen, prostate gland and seminal vesicles removed for weighing and histology. In addition, estrogen receptors were measured in the thymus. The thymus was enlarged after orchidectomy and greatly restored in aging rats. In aging rats, both aromatase inhibitors restored the thymus, which appeared normal histologically. In addition, ATD enlarged the thymus in young intact animals. Doses of testosterone which restored the accessory sex organs to weights measured in intact rats prevented the effects of orchidectomy on the thymus, and in old rats the effects of testosterone were blocked by ATD in both thymus and spleen. Available cytosolic estrogen receptors were reduced in thymus of testosterone-treated orchidectomized rats, and this effect blocked by ATD, which itself was apparently able to induce estrogen receptors. Receptors could not be detected in thymus from aging rats, but were measureable in cytosols from thymus of orchidectomized or ATD-treated old rats. It is therefore possible to restore the thymus in intact aging rats without recourse to surgical or chemical castration, and such a maneuver may possibly be of use to enhance an immune system weakened by aging or disease.


Methyl inosine monophosphate: a potential immunotherapeutic for early human immunodeficiency virus (HIV) infection. MIMP is a new thymomimetic purine under development for immunorestorative therapy. Lymphocytes were obtained from eight patients with acquired immunodeficiency disease (AIDS), eight with symptomatic pre-AIDS (ARC), and 22 normal controls and were stimulated in vitro with phytohemagglutinin (PHA). AIDS patients (mean CD4 counts of 40) showed PHA responses less than 10% of control while ARC patients (mean CD4 counts of 544) showed responses approximately 50% of the control responses. MIMP (0.1, 1, 10 and 100 micrograms/ml) progressively augmented the PHA responses in all these groups. The augmentation of the responses of the leukocytes of AIDS patients while statistically significant was minimal. The augmentation of the responses of ARC patients was significant and their maximal responses approached control levels. The effect of 1 micrograms/ml MIMP was comparable with that observed with indomethacin (10(-6) M) and interleukin-2 (IL2 - 4 units/ml) and was additive with each of these stimulants. In a parallel manner, MIMP restored the suppression of control lymphocytes induced by the immunosuppressive 17 amino acid fragment of the P41 peptide of HIV. In vivo experiments showed that MIMP significantly delayed death in a murine FLV AIDS model at a dose of 1 mg/kg by the oral or parenteral route. MIMP is under preclinical development for early HIV disease to forestall progression to AIDS by attenuating virus-induced immunosuppression.


Interleukin-6 (IL-6) in adjuvant arthritis of rats and its pharmacological modulation. The levels of serum interleukin-6 (IL-6) were measured during the course of adjuvant arthritis (FAA) in male Lewis rats. In the course of the disease serum IL-6 levels increase with a clear correlation with morphologic disease signs. Additionally, the FAA-inducing antigen, heat-killed Mycobacterium tuberculosis, was found to be a strong inducer of IL-6 production by spleen cells in vitro. The effects of the anti-rheumatic drugs indometacin, dexamethasone, cyclophosphamide and cyclosporin A (CsA) on IL-6 levels during FAA were determined. Complete normalization of serum IL-6 levels was observed with dexamethasone, cyclophosphamide and CsA whereas indometacin only partly reduced serum IL-6 levels.


Potentiation of HIV envelope glycoprotein and other immunogens by endotoxin (ET) and its molecular fragments. The structural requirements for the immunopotentiating (adjuvant) effect of endotoxin (ET) were investigated. Mild hydrolysis (0.2 N acetic acid at 95 degrees C) was applied to various ET preparations and the lipid rich (Lipid A) and polysaccharide-rich (PS) preparations obtained were tested as adjuvants on three immunogens: sheep red blood cells (SRBC), L-glutamine: L-lysine: L-alanine containing random synthetic polypeptide (GLA-40), and recombinant HIV viral envelope polypeptide (CBre3). It was found that not only the Lipid A precipitates, but under certain hydrolytic conditions the non-toxic PS preparations were also potent adjuvants. The exact conditions of hydrolysis which led to the isolation of immune adjuvant bacterial products were established. These materials were also tested for endotoxicity (Limulus lysate clotting, chick embryo lethality and local Shwartzman skin reactivity), as well as for TNF generating activities. It was found that TNF generation runs parallel with toxicity of the samples, but it does not follow the adjuvant activity of the isolates. Chemical analysis of the preparations indicated that they did not contain residual ET or Lipid A, however, they did not exclude that deacylated and dephosphorylated skeletal remains of ET are among those components in these preparations which have immunomodulatory activity.


Tunicamycin inhibits function and expression of the high-affinity IL-2 receptor in a murine IL-2-dependent cell line. Murine interleukin-2-dependent T-lymphocytes (CT6) were treated with tunicamycin, an inhibitor of both glycoprotein and ganglioside synthesis, to study the involvement of glycosylation in the IL-2 proliferative response. Tunicamycin inhibited proliferation in a dose-dependent manner at concentrations which did not inhibit protein synthesis (10-50 ng/ml). Swainsonine, a glycoprotein processing inhibitor, had no effect on proliferation. Inhibition of proliferation by tunicamycin was accompanied by an inhibition of binding of 125I-IL-2 to its high-affinity receptor. Scatchard analysis showed that receptor number was decreased by tunicamycin treatment. On the other hand, tunicamycin did not affect either the binding of the monoclonal antibody 7D4, specific for the 55 kDa low-affinity protein subunit of the IL-2 receptor, or the recycling of the IL-2 receptor. To determine the specific effects of tunicamycin on the biosynthesis of particular CT6 glycoconjugates, cells were radiolabeled with 3H-glucosamine and incorporation into ganglioside, neutral glycolipid and glycoprotein fractions was measured. Low doses of tunicamycin inhibited ganglioside synthesis and glycoprotein glycosylation to the same extent, whereas no effect on neutral glycolipid synthesis was observed. These results suggest that glycosylation of glycoprotein and/or gangliosides might play an important role in the formation of a functional high-affinity IL-2 receptor complex in CT6 cells.


The synergistic immunosuppressive potential of cyclosporin metabolite combinations. Out of the 29 cyclosporin (CS) metabolites defined so far seven representatives were isolated from the bile of liver grafted patients, purified by HPLC and characterized by FAB-MS and/or 1H-NMR. These were used to determine the growth inhibitory effects on concanavalin A stimulated rat lymphocytes (LN). Metabolites diluted in culture medium at concentrations re-checked by HPLC at the respective assay time were added and proliferation determined by [3H]-thymidine incorporation after 48 h. A 50% growth inhibition of LN by single metabolites (AM) was achieved at the following concentrations (mg/l): CS: 0.023; primary metabolites AM1: 0.11; AM1c: 0.65; AM9: 1.05; secondary metabolites AM19: 1.02; AM4N9: 1.02; H355: 1.85; AM1A: 4.5. Although all metabolites were immunosuppressive at higher concentrations in vitro on a single metabolite level, only AM1 with 20% of the activity of native CS seemed to play a role in vivo. However, when we tested the antiproliferative effects of double or triple metabolite combinations, we found a strong synergism not only of primary metabolites, but even with combinations including secondary metabolites. The concentration of the participating metabolites necessary to decrease LN growth by 50% was far below the trough levels observed in vivo. Finally, to mimic to some extent the in vivo situation we determined the interaction of native CS with single metabolites or double combinations. In contrast to the clear synergism in the absence of CS the combinations of metabolites with native CS resulted in an additive growth inhibition. These results indicate an immunosuppressive potential of all metabolites tested and a clear synergism of metabolites in the absence of CS. Although up to double metabolite combinations did only additively enhance CS induced immunosuppression, the combination of 29 metabolites occurring in vivo might have significant immunosuppressive effects in situations where CS levels drop below active concentrations.


Immunomodulating properties of carbamazepine in mice. The protective effects of carbamazepine (CBZ) were studied in mice inoculated with Lewis Lung Carcinoma (3LL), Madison Lung Carcinoma (M109), L5178Y lymphoma, L1210 leukaemia and Candida albicans. There was no significant increase in survival time of mice treated with CBZ. However, CBZ, as well as its metabolite CBZ 10-11 epoxide (CBZ 10-11 EPOX), showed a significant increase in NK-cell activity. CBZ also produced a significant increase of phagocytosis and killing properties of PMNs. There was no significant difference in the stimulation of splenic lymphocyte blastogenesis by different concentrations of phytohaemagglutinina (PHA), observed between the controls and CBZ treated mice. The results demonstrate that the effect of chronic treatment with CBZ on the immune response is a complex phenomenon which remains a challenge for future research.


The immunomodulator AS101 administered orally as a chemoprotective and radioprotective agent. AS101 [ammonium-trichloro (0,0' dioxyethylene)tellurate] is a new immunomodulator shown previously to stimulate the production of various cytokines in vitro and in vivo, and to have minimal toxicity. In the present study we explore the possibility of oral administration of AS101 to mice via cannulation in lieu of interperitoneal or intravenous administration reported to date. Our studies show that oral administration of AS101 at a dose ranging between 50 and 100 micrograms/mouse promotes hemopoietic regeneration after treatment with sublethal doses of cyclophosphamide (CYP) and protects mice from the lethal effects of this compound. In addition, AS101 administered orally confers a strong radioprotective effect upon mice when given before irradiation.


The interleukins-1 alpha, -1 beta, and -2 do not acutely disrupt the murine blood-brain barrier. Previous studies have suggested that some of the central nervous system (CNS) effects of interleukin-2 (IL-2) and perhaps other cytokines might be mediated through disruption of the blood-brain barrier (BBB). We investigated the ability of human IL-2 and, in selected studies, human IL-1 alpha and human IL-1 beta to disrupt the BBB to radioiodinated bovine serum albumin (RISA) after intravenous (i.v.) and intracerebroventricular (i.c.v.) injection. No disruption of the BBB occurred for up to 2 h after the i.v. injection of 2 micrograms/mouse of IL-2 (10(5) U/kg of body weight), 2 micrograms of IL-1 alpha (10(7) U/kg), or 2 micrograms of IL-1 beta (10(7) U/kg). This dose of i.v. IL-2 also did not affect BBB permeability to RISA in the brain to blood direction. Damage to the BBB induced by hypertension elicited by i.v. epinephrine was not enhanced or prolonged by IL-2. When given directly into the CNS by the i.c.v. route, 100 ng of IL-2 (2.2 x 10(5) U/kg of brain), 100 ng of IL-1 alpha (2.2 x 10(7) U/kg of brain), or 100 ng of IL-1 beta (2.2 x 10(7) U/kg of brain) had no effect on BBB integrity in either the blood to brain or the brain to blood direction. We conclude that the effects of IL-1 alpha, IL-1 beta, and IL-2 on the CNS, as studied under these conditions, are not due to disruption of the BBB but are mediated by other mechanisms including the ability of some interleukins to cross the BBB by a saturable transport system described previously.


Hydrolysis and association of leucine enkephalin to lymphomic and erythroleukaemic cell lines--II. The relationship between the hydrolysis of labelled leucine enkephalin and the association of its radioactive label to the cells of lymphomic and erythroleukaemic cell lines have been studied using intact cells and resealed membranes obtained from these cells as models. Hydrolysis by cell enzymes and its effect on association have been analysed using protease inhibitors and non-hydrolysable enkephalin analogues. Results obtained confirm that hydrolysis of the pentapeptide is a prerequisite for association of the radiolabel to cells. The same results provide evidence of marked differences between enkephalin hydrolysis by whole cells and hydrolysis measured in the presence of resealed membranes, suggesting the existence in intact cells of proteolytic enzymes other than those bound to the membranes. The lack of reversibility of association and the intracellular localization of the radioactive label suggest that the association measured is prevailingly caused by internalization of a hydrolysis fragment, and not by binding to receptors. In order to determine the nature of the active fragment, association was measured in the presence of all four labelled N-terminal hydrolysis fragments of leu-enkephalin under conditions of nearly-total inhibition of proteolytic enzymes. Under these conditions, the label carried by Tyr, but not that carried by the other N-terminal fragments, was associated with cells. Free Tyr, furthermore, inhibits the association to cells of both labelled Tyr and leu-enkephalin. Data summarized above are consistent with the hypothesis that the radioactive label is taken up by the cells as Tyr, freed from the parent peptide by cell-related enzymes. The same data tend to exclude that a relevant fraction of the intact pentapeptide is bound to membrane receptors or that the radioactive label is carried into the cell by a N-terminal fragment other than Tyr.


Modulation of colony-stimulating activity by interleukin 1 in mice: opposing effects of combined treatment with indomethacin or prostaglandin E2. Following previous observations that interleukin 1 (IL-1) may have both positive and negative effects on the levels of circulating colony-stimulating factors (CSF) in mice, we have investigated the impact of human rIL-1 beta administration on serum concentrations of colony-stimulating activity (CSA, as defined by biossay) and macrophage-specific colony-stimulating factor (CSF-1, measured by RIA). In addition, we have studied the effects of IL-1 administered in conjunction with indomethacin or prostaglandin (PG) E2. Besides confirming the finding that exogenous IL-1 leads to a rapid increase in CSF detection, we obtained evidence that IL-1 may also result in the production of cyclo-oxygenase pathway products that down-regulate the IL-1-induced burst in CSA and CSF-1 levels. While co-treatment of mice with indomethacin led to a further increase in CSF detection, the combined exposure to IL-1 and PGE2 resulted in a significant impairment of the stimulatory activity of IL-1.


Drug-specific immune responses induced by procainamide, hydralazine and isoniazid in guinea-pigs. The drug-induced graft vs host reaction (GVHR) hypothesis requires, as its first step, specific T-cell immune responses to the drug-modified self. Procainamide, isoniazid and hydralazine are known to provoke various allergic reactions including GVHR-like adverse effects in man. We now report that drug-specific immune responses can easily be induced by these drugs in guinea-pigs. Twenty-five milligrams of each of these drugs and penicillin G, which is known to make covalent bonds with proteins and to also induce drug-specific immune responses, were mixed with complete Freund's adjuvant (CFA) and subcutaneously (s.c.) injected twice at an interval of 2 weeks into female Hartley guinea-pigs. The antibodies to these drugs were assessed by means of an enzyme-linked immunosorbent assay (ELISA). Two weeks after the last injection, all animals treated with isoniazid, hydralazine and penicillin G produced high titers of antibodies to these drugs. Antibodies to procainamide were also detected, although their antibody titers were low. The specificity of the antibodies produced were tested by the inhibition of ELISA and concentration-dependent inhibition was observed. Delayed type hypersensitivity (DTH) reactions were also observed in the animals treated with procainamide, isoniazid and hydralazine 2 weeks after the last injection. These results suggest that the allergic reactions observed in clinical use are related to the inducing potential of drug-specific immune responses in an animal system. Therefore, immunization of guinea-pigs with test drugs and CFA may give useful information for predicting the occurrence of allergic reactions in man.


Popliteal lymph node enlargement induced by procainamide. The ability of procainamide (PA) to induce primary local popliteal lymph node (PLN) reactions has been investigated. We employed an in vitro drug-metabolizing system, based on the hypothesis that the negative result for PA in a PLN assay (PLNA) was due to insufficient metabolizing activity at the reaction site. PA was incubated previously in vitro with the S-9 mixture derived from rat liver. The reactants were ultrafiltered in order to eliminate high molecular-weight molecules, and then the low molecular-weight fraction was subcutaneously (s.c.) injected into the hind footpad of mice. PLN reactions were assessed by weighing the popliteal lymph node of the injected side. The reactants of more than 5 mg of PA and S-9 mixture induced PLN enlargement in C3H/He mice 8 days after injection. BALB/c and C57BL/6 mice were also susceptible to PLN reaction in response to the reactants of PA (10 mg) and S-9 mixture. PLN reactions to PA were induced 2 days after the injection and sustained until 18 days. Contact of PA with the S-9 mixture for 30 min at 37 degrees C was sufficient to induce PLN enlargement. However, contact for 24 h reduced the peak reaction. On the other hand, PA which had not been incubated with the S-9 mixture and acetylprocainamide (acetylPA) gave little or no reaction. Local PNA has not been considered to be suitable for the detection of drugs with the potential to induce immune disorders in cases where a metabolite contributes to the adverse reaction. However, the employment of an in vitro drug-metabolizing system may overcome that defect.


The role of lymphocyte production and migration in the lymphopenia caused by 2-acetyl-4-tetrahydroxybutyl imidazole. 2-Acetyl-4-tetrahydroxybutyl imidazole (THI), a component of the food colouring ammonia caramel, has been shown to produce a profound and rapid lymphopenia in peripheral blood in the rat. In order to investigate whether the cause of the lymphopenia was due to the reduced production and influx in the circulation, redistribution of lymphocytes into other lymphoid compartments or an increased cell death, THI (1 mg/kg/day) was given in the drinking water for up to 14 days to F344 rats. A profound depletion of lymphocytes after already 1 day was only found in the blood compartment, whereas no such marked and rapid changes were found in the cellularity of other lymphoid compartments. The proportion and absolute number of DNA-synthesizing cells in each lymphoid organ was quantified using an antibody directed against incorporated 5-bromo-2'-deoxyuridine (BrdU), 1 h after a single BrdU injection. Additionally, enumeration and localization of BrdU+ cells was determined at later time points after a single BrdU injection by flow cytometry and immunocytochemistry, in order to examine the distribution and localization of recently formed (BrdU+) lymphocytes. THI treatment had no effect on the proliferation rate and the distribution of newly formed (BrdU+) cells in the lymphoid organs. However, migration studies revealed that THI treatment resulted in an increased percentage of fluorescein-labelled peripheral blood lymphocytes found in the spleen and bone marrow and a decreased percentage in the cervical and mesenteric lymph nodes, 24 h after injection. Collectively these results indicate that the lymphopenia in the peripheral blood compartment after THI treatment, is caused by a rapid sequestration of lymphocytes into the spleen and bone marrow rather than by a reduced lymphocyte production and release into the periphery. The fact that THI also caused lymphopenia in splenectomized rats, indicates that the spleen does not play an active part in the change in migrational behaviour of lymphocytes after THI treatment. Finally, as there was no increase in the absolute number of lymphocytes found in the spleen or bone marrow it seems they are rapidly degraded.


2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) inhibits the activation of antigen-specific T-cells in mice. There are conflicting data in the literature regarding target cells in 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-induced immunotoxicity. In the present study, adult male C57BL/6 mice were exposed to TCDD (50 micrograms/kg) 4 days prior to immunization with ovalbumin (OVA). The effect of TCDD on the specific immune response in vivo was determined by T-cell proliferation and IL-2 production in response to either OVA or anti-mouse-CD3 antibodies plus PMA in vitro. The antigen-specific T-cell proliferation and IL-2 production in response to OVA were significantly suppressed by TCDD, while the polyclonal response to anti-CD3 antibodies plus PMA was not affected. This indicates that even at a high dose of TCDD the intra T-cell signalling pathways in resting cells are not disturbed, but TCDD selectively impairs the antigen-specific activation of T-cells. Since activated T-cells are required in antibody responses to T-dependent antigens, the low number of such cells observed in the present study, may well explain the suppressive effects of TCDD on humoral immunity reported previously.


Mechanism of ethanol-mediated immunosuppression in mice: ethanol suppresses T-cell proliferation without affecting IL2 production and IL2 receptor expression. The effect of extended ethanol consumption in young C57BL/6 mice on T-cell proliferation was studied. Splenic cells of young mice (3-4 months old), fed with one of three different liquid diets (5% ethanol, maltose-substitute, or standard liquid diet) for 28-38 days were cultured with plant lectins to assess T-cell proliferation and IL2 production. Expression of T-cell subset markers (CD4+/CD8+) was also determined. Then, Con A-activated T blast cells were assessed for their ability to express IL2 receptor (IL2R) and to respond to IL2. Finally, the proliferative response of splenic cells to PMA/ionomycin was assessed. The results showed that both lectin- and PMA/ionomycin-induced mitogenesis and IL2-dependent proliferation of T-cells from ethanol diet-fed mice were diminished as compared with that of maltose-substitute diet or standard liquid diet. However, the ability of T-cells from ethanol diet-fed mice to produce IL2 and to express IL2 R or CD4+/CD8+ subset markers was not affected. Furthermore, the magnitude of ethanol-mediated suppression of T-cell proliferation induced by PMA/ionomycin was comparable with that induced by Con A. These results taken together indicate that ethanol suppresses T-cell proliferation by interfering with events following the IL2-IL2R interaction. Therefore, it is likely that ethanol inhibits murine T-cell proliferation by selectively affecting the progression (IL2R-mediated events) rather than the initiation (mitogenic receptor-mediated events) of the cell cycle.


Changes in plasma lipoprotein levels during a hiking expedition in South America. Eleven males participated in a hiking expedition over a period of 6 weeks during which they walked an average of 15 km per day, resting days included. The participants completed a seven-day estimated dietary record before and during the expedition. Their habitual dietary intake before the expedition was typical of a Western diet. During the expedition most animal products, with the exception of canned fish, were excluded from the diet. The dietary intake of fat and carbohydrate changed from 36.9% and 40.6% to 14.0% and 76.4% respectively. Cholesterol intake dropped from 557 mg to 92 mg. Mean plasma total cholesterol (TC) decreased from 190.9 mg/100 ml to 142.0 mg/100 ml. These changes were mainly due to changes in low density lipoprotein cholesterol (LDL-C). Although high density lipoprotein cholesterol (HDL-C) did not change, the ratio of HDL-C to TC increased significantly. It can be concluded that drastic dietary changes, together with increased physical activity and weight loss resulted in major plasma lipoprotein changes. The expected fall in HDL-C due to a high carbohydrate diet was counteracted by the increased physical activity and weight loss.


Fluid replacement after dehydration: influence of beverage carbonation and carbohydrate content. This investigation evaluated the effects of beverage carbonation and carbohydrate (CHO) content on fluid replacement following exercise/thermal dehydration. On four occasions separated by at least 7 days, eight healthy men cycled at 50% of VO2max in a hot environmental chamber (40 degrees C, 40% relative humidity) until a weight loss of 4.12 +/- 0.22% was attained. In the subsequent four hours, subjects ingested one of four solutions at 15-min intervals. The total volume ingested equalled that lost during dehydration. The solutions were administered in randomized order and varied in their carbonation and carbohydrate (CHO) content: 1. CK: carbonated 10% glucose-fructose solution, 2. NCK: non-carbonated 10% glucose-fructose solution, 3. CNK: carbonated non-caloric solution, and 4. NCNK: non-carbonated non-caloric solution. Plasma volume changes, total plasma protein concentration, plasma osmolality, and the plasma glucose concentration were determined at rest before and after dehydration, and at 30, 90, 150, and 240 min of recovery. Plasma volume changes and the plasma protein concentration were not different (p greater than 0.05) between treatments. Values for the plasma glucose concentration and the change in plasma osmolality were significantly elevated when CHO beverages were ingested when compared with non-CHO beverage ingestion. Five-min cycling bouts were performed at 70% of VO2max before and after dehydration and at 60, 120, 180, and 240 min of rehydration. The respiratory exchange ratio was elevated in both of the CHO treatments when compared with both of the non-CHO treatments at 60, 120, 180 and 240 min of rehydration.(ABSTRACT TRUNCATED AT 250 WORDS)


Depressed systolic and diastolic cardiac function after prolonged aerobic exercise in healthy subjects. We studied 11 healthy untrained volunteers (aged 28.9 +/- 4.6 years) during 60 minutes of aerobic ergometric exercise with constant heart rates of 130 to 140 beats/minute. We found a continuous and significant decrease in systolic and diastolic pressure from 175 +/- 18/77 +/- 7 mmHg in the 5th minute to 144 +/- 14/68 +/- 6 mmHg in the 60th minute of exercise. Cardiac function and structure were assessed by M-mode echocardiography before exercise, after 5 minutes and after 60 minutes of exercise at comparable heart rates. The results demonstrated significant decreases in cardiac output, ejection fraction, and diastolic posterior wall velocity and an increase in total peripheral resistance after 60 minutes of exercise. We conclude that the decrease in blood pressure during long-term aerobic exercise in healthy untrained subjects might be at least influenced by a decrease in left ventricular filling and contractility, possibly indicating cardiac fatigue.


Bioenergetic characteristics of swimmers determined during an arm-ergometer test and during swimming. The maximal oxygen uptake (VO2max) of 13 swimmers was determined by an arm-ergometer test (direct method) and estimated from a maximal multistage swimming test (indirect method) (23). A test-retest of the progressive swimming exercise showed that there were no significant differences from one test to the other and that there were significant correlations between the principal parameters: arm stroke index: 0.73, maximal aerobic swimming velocity: 0.94, VO2max: 0.95, p less than 0.01. Therefore, for swimmers of average ability, the reproducibility of this test has been proved. A significant difference (p less than 0.001) was observed between the two tests for VO2max: arm-ergometer test (VO2max arms): 2.4 +/- 0.5 l.min-1, swimming test (VO2max ST): 3.2 +/- 0.7 l.min-1, p less than 0.01. This difference appeared to be linked to the use of a greater muscle mass (arms and legs) during swimming. A significant correlation (r = 0.73, p less than 0.01) was obtained between VO2max (l.min-1) by using both the direct and indirect exercises as methods of measurement. However, the level of r did not permit the prediction of one parameter from the other. Significant correlations were obtained between VO2max and performances over 200 and 400 m free style regardless of the methodology used (VO2max arm, VO2max ST). Moreover, only VO2max (arm, ST) emerged as a variable accounting for swimming performance from a step-wise multiple regression analysis, in which biometric and bioenergetic parameters were taken into account.


A simple method for gender verification based on PCR detection of Y-chromosomal DNA and its application at the Winter Universiade 1991 in Sapporo City, Japan. A simple and reliable screening method for gender verification at international sports competitions was developed on the basis of detection of Y-chromosomal DNA which was specifically amplified by polymerase chain reaction (PCR). Total DNA obtained from buccal mucous membrane cells by digestion with proteinase K followed by boiling treatment was directly used for the PCR. The amplified product was electrophoresed on an agarose gel staining with ethidium bromide. We adopted three sets of the PCR primer, identifying different regions of Y-chromosome, of which sensitivity and specificity were preliminary evaluated for 228 (112 male, 116 female) DNA samples from lymphocytes. This new method was first applied for the gender verification test at the Winter Universiade 1991 Sapporo, accompanied by traditional microscopic tests for X- and Y-chromatins. No positive results were obtained for 155 female competitors using both the PCR and the microscopic methods. The superiority of the proposed method was clearly shown in the reliability of the results and also in the saving on instrumental and personnel costs as compared to previous cytologic methods.


Effects of captopril chronic intake on the aerobic performance and muscle strength of normotensive trained subjects. Fifteen normotensive athletes specializing in dynamic sports took part in a randomized double-blind and cross-over study: captopril (50 mg/24 h) vs placebo. Each treatment lasted one month. Maximal exercise tests on cycle ergometer were performed at the end of each period. No significant differences were observed in the maximal values of oxygen uptake, power, heart rate or blood lactate value. The anaerobic threshold, defined as the exercise intensity which corresponded to a 4 mmol.l-1 blood lactate level was unchanged. With captopril, the end-of-exercise systolic and diastolic BP were slightly altered (NS). Lower limb muscle strength, as explored with a Cybex isokinetic system, was not modified by captopril intake. The results indicate that maximal aerobic performance and isokinetic strength of the lower limbs are not altered by captopril chronic administration (50 mg/day) in normotensive trained subjects.


Effect of training on the calf muscle energy metabolism. A 31P-NMR study on four elite downhill skiers challenged with a standardized exercise protocol. This study evaluates the effects of a 6 months' training period on the bioenergetics of the calf muscle of elite athletes by means of phosphorus-31 Nuclear Magnetic Resonance Spectroscopy (31P-NMR). Four downhill skiers, belonging to the Spanish National Team, performed a standardized exercise protocol using their right leg inside a wide-bore 2.35 Tesla magnet. The inorganic phosphate/phosphocreatine (Pi/PC) ratio and intracellular pH (pHi) were measured at steady-state during an exercise protocol composed of 5 work levels between 20% and 80% of the maximum voluntary contraction (MVC), before and after the training period. The measured values, which were markedly scattered at the beginning, regrouped after training. This was caused by a shift towards lower Pi/PC ratios and by a lower pHi acidification in three of the four subjects. This result suggests that 31P-NMR is a good tool to evaluate changes in the muscle aerobic capacity of athletes induced by training.


Body fat estimations by electrical impedance and infra-red interactance. The purpose of this study was to examine body fat estimation using three methods of electrical impedance (the BIA-103 Body Composition Analyser--RJL Systems, Detroit; the BMR 2000 Body Composition Analyzer--Berkeley Medical Research, San Leandro; the BC300 Body Composition Analyzer--Spacelabs, Dallas) and an infra-red interactance method (Futrex 5000 Analyzer--Futrex Inc. Gaithersburg) as an alternative to hydrodensitometry. Five different groups were examined using at least one of the electrical systems and in all cases utilising hydrodensitometry as the criterion method. The results produced highly significant correlations between all methods, but caution is recommended due to the limited common variance in some cases. The individual electrical methods differed from hydrodensitometry by a maximum of 1.1% in obese women, by 21.6% in athletic adults, by 6.2% in the slightly obese group, by 8.1% in normal women and by 56.0% in normal children. Significant mean differences between one of the impedance methods and hydrodensitometry were only noted in two of the groups tested. This suggests, allowing for the limitations of hydrodensitometry itself, that the other three electrical methods appear to be reasonably valid alternatives to underwater weighing, especially for field work of an epidemiological nature.


Cortisol levels during prolonged exercise: the influence of menstrual phase and menstrual status. The purpose of this study was to determine the influence of menstrual phase and menstrual status on the cortisol response during 90 minutes of treadmill running at 60% VO2max. Eight eumenhorrheic athletes were tested in the early follicular (EF) (day 3-5), late follicular (LF) (day 13-15) and mid-luteal (ML) (day 22-24) phases. Six amenorrheic athletes were tested on two separate occasions. The resting cortisol levels were similar in each menstrual phase and overall a decreasing pattern of cortisol response to exercise was observed in all menstrual phases (P greater than .05). The amenorrheic athletes had a significantly greater (P less than .01) pattern of cortisol response than was observed in eumenorrheic athletes. The net increment in cortisol levels during exercise were distinctly greater (P less than .01) in amenorrheic than eumenorrheic athletes (amenorrheic: 413.8 +/- 113.1, eumenorrheic: EF: -482.8 +/- 88.3, LF: -311.8 +/- 102.1, ML: -386.3 +/- 146.2 nmol.l-1). In conclusion the cortisol levels are independent of menstrual phase. Also a larger cortisol increment is observed in amenorrheic athletes in response to prolonged submaximal exercise. The elevated cortisol levels in amenorrheics at rest and throughout exercise provides further evidence that disturbances in the hypothalamic-pituitary-adrenal function are associated with exercise-induced amenorrhea, although the site(s) of physiological disturbance have not been identified.


Biochemical, hematological and endocrinological parameters during repeated intense short-term running in comparison to ultra-long-distance running. In order to see the differences in the acute effects of two kinds of running on biochemical, hematological and hormonal parameters, we investigated in one 24 h race (same exterior conditions) two groups of runners (NR = nonstop runners; RR = relay runners). From each participant blood samples were taken before, 3 times during and after the race. In both groups the lipid parameters cholesterol, triglycerides and LDL-cholesterol declined significantly, triglycerides even to 40% of their initial value, whereas HDL-cholesterol increased significantly. Renal parameters showed similar increases in both groups except for uric acid, which rose by 73% in RR compared with only 23% increase in NR. A difference was also found in the lactate and glucose concentrations, showing a four-fold increase and a 34% increase at 4 h respectively in RR, whereas the NR remained constant. Changes in the hematological parameters were similar in both groups. Cortisol and prolactin showed similar alterations during the race but LH and testosterone declined to approximately half of their initial concentration (p greater than 0.05) in NR after 4 to 8 h, in contrast to RR, where the concentration of LH or testosterone remained constant or changed only moderately but insignificantly during the race. Our investigation indicates that in ultra-long-distance running and intense short-term exercise performed under the same exterior conditions, many parameters show similar changes but great differences can be found as well if certain hormones, glucose metabolism and renal parameters are considered.