[Studies on the effects of rhTNF-alpha anti-replication against EHFV and HAV in vitro]. It was demonstrated in this study that rhTNF-alpha can inhibit the replication of epidemic hemorrhagic fever virus and hepatitis A virus in tissue culture. EHFV infection produced strong fluorescence ( - +) on Vero-E6 cells after 9 days. In the rhTNF-alpha treated group, however the fluorescence was very weak (- or +/-). rhTNF-alpha also showed anti-HAV effects on 2BS cells. The fluorescence intensity on 2BS cells infected by HAV was - +, but in the rhTNF-alpha treated group, the fluorescence was negative. We observed that rhTNF-alpha inhibited EHFV antigen expression on Vero-E6 at different times (-2h, 0h, +2h). Dr. Old had recently suggested that the physiological role of rhTNF-alpha is that of a protective agent against infectious diseases, and this view is supported by our findings.


[PCR amplification, cloning and expression of the gene coding for CD4 V1-V2 domains]. We describe the use of a modified technique of polymerase chain reaction (PCR) for facilitating cloning and expression of a cDNA fragment encoding CD4 V1-V2 domains. The modification includes introducing suitable signals to start primer and halt primer of the target gene as indicated in Fig. 1. After verified by DNA sequencing, the amplified DNA fragment was cloned into prokaryotic expression vectors containing lambda PL promoter. The screened clones were induced to express the target fragment V1-V2 of CD4 gene and the expected gene product was estimated to be 8% or 10% of the total cellular proteins.


Sympathetic and immune interactions during dynamic exercise. Mediation via a beta 2-adrenergic-dependent mechanism. The relation between the sympathetic nervous system and the immune system has not been fully defined. Recent investigations have suggested an adrenergically driven efflux of specific beta 2-receptor-rich lymphocyte subsets into the circulation with either exercise or infusion of exogenous catecholamines. To determine whether acute sympathetic stimulation mediates immunoregulatory cell traffic and function via a beta 2-receptor mechanism, we exercised 20 healthy volunteers before and after 1 week of treatment with either the nonselective beta-antagonist propranolol or the beta 1-selective antagonist metoprolol. Before treatment, exhaustive exercise according to the Bruce protocol led to a marked lymphocytosis. Tsuppressor/cytotoxic (Ts/c) and natural killer cells, subtypes with the largest density of beta-receptors, showed the most pronounced increases after exercise, with less impressive elevations in T(helper) and B cells. With respect to function, exhaustive exercise led to a decrease in concanavalin A-stimulated IL-2 receptor expression and [3H]thymidine incorporation while enhancing natural killer cell activity. One week of propranolol therapy blunted the exercise-induced increases in circulating Ts/c and natural killer subpopulations as well as the previously observed alterations in cellular immune function. Treatment with the beta 1-selective antagonist metoprolol, however, did not impair the influence of exercise on any of the above parameters. Acute sympathetic stimulation by exhaustive exercise leads to selective release of immunoregulatory cells into the circulation with subsequent alterations in cellular immune function, either secondary to subset changes or as a result of direct catecholamine effects on function. These changes are attenuated by propranolol but not metoprolol, suggesting a beta 2-mediated mechanism.


The role of L-arginine in ameliorating reperfusion injury after myocardial ischemia in the cat. Myocardial ischemia followed by reperfusion results in endothelial dysfunction characterized by a reduced release of endothelium-derived relaxing factor (EDRF). Because EDRF has been characterized as nitric oxide, we examined the ability of L-arginine, the substrate for nitric oxide synthesis, to protect in a feline model of myocardial ischemia plus reperfusion. The effects of L-arginine were investigated in a 6-hour model of myocardial ischemia and reperfusion in pentobarbital-anesthetized cats. A bolus administration (30 mg/kg) of L-arginine, or its enantiomer D-arginine, was given followed by a continuous infusion of 10 mg/kg/min for 1 hour starting 10 minutes before reperfusion. Myocardial ischemia plus reperfusion in cats receiving D-arginine resulted in severe myocardial injury and endothelial dysfunction characterized by marked myocardial necrosis, high cardiac myeloperoxidase activity in ischemic cardiac tissue, and loss of acetylcholine- and A-23187-induced endothelium-dependent relaxation in coronary artery rings. In contrast, myocardial ischemia plus reperfusion cats treated with L-arginine exhibited a reduced area of cardiac necrosis (16 +/- 2% versus 41 +/- 5% of area at risk, p less than 0.01), lower myeloperoxidase activity in the ischemic region (0.3 +/- 0.08 versus 0.8 +/- 0.10 units/100 mg tissue, p less than 0.05), and significant preservation of acetylcholine- (p less than 0.01) and A-23187- (p less than 0.01) induced endothelial-dependent relaxation. These results demonstrate the ability of L-arginine to reduce necrotic injury in a cat model of myocardial ischemia plus reperfusion, and this reduction in infarct size is associated with the preservation of endothelial function and attenuation of neutrophil accumulation in ischemic cardiac tissue.


Noninvasive assessment of cardiomyopathy development with simultaneous measurement of topical 1H- and 31P-magnetic resonance spectroscopy. Background. It might be possible to estimate the metabolic derangement of cardiac muscle by topical nuclear magnetic resonance spectroscopy (MRS) in vivo without killing the animal. Methods and Results. By use of topical 1H- and 31P-MRS focused on the heart of Syrian hamsters with or without cardiomyopathy (CM; BIO 14.6 strain), the chemical constituents were measured in vivo nondestructively and repetitively at several stages of development of CM. A phantom experiment and two-dimensional plot of chain methylenes (CH2) of lipid/water ratio by 1H-MRS versus creatine phosphate (CP)/[beta-P]ATP ratio by 31P-MRS indicated that signal cross talk from the adjacent organs was negligible. Even before the onset of clinical or pathological manifestation of CM (7 weeks after birth), CH2/water ratio by 1H-MRS was lower in the CM group (7.3 +/- 0.7%) than in control (11.8 +/- 2.0%, p less than 0.05), and it decreased further at the hypertrophic stage (17 weeks, 4.1 +/- 0.7%, p less than 0.05) and the congestive stage (27 weeks, 4.3 +/- 0.9%, p less than 0.05). In contrast, the CP/[beta-P]ATP ratio by 31P-MRS started to decrease at the hypertrophic stage (1.90 +/- 0.18 versus 2.52 +/- 0.24, p less than 0.05) and decreased further at the congestive stage to 1.53 +/- 0.18 (p less than 0.01). These in vivo MRS data were confirmed by both biochemical assay and in vitro MRS analysis in heavy water after the animals were killed. Conclusions. A combination of topical 1H-MRS and 31P-MRS in vivo is promising for the noninvasive and sensitive assessment of cardiac muscle metabolism. Comparison of these MRS studies and biochemical analysis suggested not only the modification of water, lipid, CP, or ATP contents but also the reduction of flexibility or fluidity of lipids in cardiomyopathic heart.


Hypothalamic abnormalities in patients with pituitary-dependent Cushing's syndrome. We aimed to investigate the pattern of 24-hour ACTH and cortisol secretion in pituitary-dependent Cushing's syndrome and to evaluate the pituitary and hypothalamic contributions to the disease. Five women with Cushing's disease (mean age 35 +/- 5 (SEM) years) and five normal female controls (mean age 25 +/- 2 years) were studied. Plasma ACTH and cortisol levels were measured every 15 minutes for 24 hours using established IRMA and RIA respectively. ACTH and cortisol mean and trough levels, pulse number and amplitude were calculated using established computer software, programmed to identify ACTH and cortisol peaks. Patients with Cushing's disease had a twofold increase in 24-hour mean cortisol levels and a threefold increase in 24-hour mean ACTH levels (Cushing's 5.9 +/- 1.0, controls 1.9 +/- 0.2 pmol/l, P less than 0.01). This was predominantly mediated by an increase in ACTH pulse amplitude. However, 24-hour ACTH pulse number was also increased (Cushing's 15.2 +/- 2.6, controls 10.6 +/- 1.7, P less than 0.05) due to an increase in pulse number between 1800 and 2400 h. ACTH trough levels were also higher in patients with Cushing's disease (Cushing's 5.3 +/- 1.3, controls 2.3 +/- 0.2 pmol/l, P less than 0.05). Twenty-four-hour mean plasma cortisol and ACTH levels are elevated two to three-fold in patients with Cushing's disease. The increase in ACTH pulse amplitude suggests a pituitary abnormality in patients with Cushing's disease. However, the increased ACTH pulse frequency together with elevated trough levels is interpreted as indicating coexisting hypothalamic stimulation (or loss of inhibition).


Correlation between spontaneous metastatic potential and type I collagenolytic activity in a human pancreatic cancer cell line (SUIT-2) and sublines. A human pancreatic cancer cell line (SUIT-2) and four sublines cloned in vitro (S2-007, S2-013, S2-020 and S2-028) were inoculated into nude mice for assessment of metastatic potentials. After 16 weeks of subcutaneous injection, the parent SUIT-2 line metastasized to the lungs and lymph nodes in three of six mice. S2-007 cells presented the highest metastatic potential in pulmonary (5/6) and lymph node (2/6) metastases among the four sublines. No metastasis was found in S2-028. The incidence of spontaneous pulmonary metastasis was correlated with that of pulmonary colonization after intravenous (i.v.) injection of cell clusters (r = 0.87, P = 0.056). Pulmonary colonization potential using single cells, however, did not always reflect a spontaneous metastatic ability. Type I collagenolytic activity in serum-free conditioned media of these cells was correlated effectively with the incidence of spontaneous pulmonary metastasis (r = 0.92, P = 0.026) and pulmonary colonization after i.v. injection of cell clusters (r = 0.95, P = 0.013). Thus, type I collagenolytic activity may possibly be essential to spontaneous cancer metastasis.


Staging lung carcinoma with a Tc-99m labeled monoclonal antibody. Thirty-three patients with biopsy-proven lung cancer and a total of 150 lesions diagnosed by conventional staging procedures were imaged using a Tc-99m labeled monoclonal Fab fragment of an IgG2B murine monoclonal antibody (MoAb) (NR-LU-10, NeoRx Corporation). Immunoscintigraphy demonstrated 100% of primary and 78% of metastatic lesions. MoAb imaging detected 88% of lesions in 12 small cell lung cancer (SCLC) patients and 77% of lesions in 21 non-small cell lung cancer (NSCLC) patients. Based on initial evaluation by other methods, 29 sites of MoAb activity were not associated with evidence of disease. Eleven of these were subsequently shown to represent sites of metastases; 18 remain unconfirmed. Four of ten patients studied with limited NSCLC had eight unsuspected lesions on MoAb imaging. Confirmation of unsuspected lesions in two patients altered initial clinical staging, and surgical therapy was abandoned. This study demonstrates that Tc-99m labeled NR-LU-10 can accurately stage patients with lung cancer.


Detection and staging of small cell lung carcinoma with a technetium-labeled monoclonal antibody. A comparison with standard staging methods. Tumor-associated radiolabeled monoclonal antibodies (MoAb) can detect neoplasms in a variety of settings. The authors conducted a study comparing the ability to detect and stage small cell lung carcinoma by using a Tc-99m labeled monoclonal antibody (NR-LU-10 Fab) (NeoRx Corp, Seattle, WA) with standard staging methods. Standard staging included a physical examination, chest x-ray, a battery of radionuclide scans and/or computerized tomographic studies (head, abdomen, and bone), and bone marrow examination. A total of 22 comparisons were performed in 17 patients (five patients had reevaluations after therapy). Fifty-four (74%) of the 73 lesions defined by standard staging were detected by the radiolabeled MoAb. Seven of eight patients (88%) classified by standard staging as having "limited stage" disease on presentation were concordantly "limited stage" by radioimmunoimaging. One patient deemed "limited stage" by standard staging was correctly upstaged (bone marrow involvement) as a result of the radiolabeled MoAb. Two patients found to have extensive disease at diagnosis were characterized as "limited stage" by the MoAb, for an overall staging accuracy of 0.88. Thirteen of 19 missed lesions were smaller than 2 cm (10 were smaller than 1 cm; 3 measured 1 to 2 cm). This comparative study shows that radioimmunoimaging by Tc-99m labeled NR-LU-10 Fab antibody is capable of complementing standard staging methods used in the evaluation of small cell lung carcinoma.


Effect of stannous pyrophosphate red blood cell gastrointestinal bleeding scan on subsequent Meckel's scan. Both labeled RBC and Meckel's scans have been used to evaluate pediatric patients with gastrointestinal bleeding, sometimes sequentially in the same patient. Particularly in infants, from whom withdrawal of sufficient blood for in vitro RBC labeling is often not possible, in vivo labeling with stannous pyrophosphate is used. However, prior administration of stannous-containing agents is known to alter the in vivo distribution of Tc-99m pertechnetate and to interfere with the subsequent Meckel's scan. The authors report on a Meckel's scan performed on an infant 1 week after a GI bleeding study with Tc-99m and stannous pyrophosphate. The Meckel's scan shows abnormal tracer distribution with absent gastric uptake, rendering the scan uninterpretable. In pediatric patients with gastrointestinal bleeding, a Meckel's scan should be done before labeled RBC imaging.


The quinolones. An overview of their pharmacology. The fluoroquinolones represent a relatively new class of antibiotics with outstanding therapeutic potential, attributable to their broad spectrum of antimicrobial activity and favourable tissue distribution. They are highly active against most Gram-negative pathogens, as well as Staphylococcus aureus and coagulase-negative staphylococci. In addition, the fluoroquinolones have useful pharmacokinetic properties: they are orally active, and their lipophilicity and low degree of plasma protein binding allow for excellent tissue penetration and concentrations, as reflected in their particularly large apparent volumes of distribution. Infections due to aerobic Gram-negative pathogens are considered those most susceptible to the quinolones. Disease indications in which these agents appear to offer the greatest therapeutic advantage over currently available alternatives include the following: complicated urinary tract infections (particularly those caused by Pseudomonas aeruginosa or resistant Gram-negative microorganisms); suspected bacterial gastroenteritis; eradication of Salmonella typhi from the faeces in known carriers; P. aeruginosa-associated respiratory exacerbation in patients with cystic fibrosis; and chronic Gram-negative bacterial osteomyelitis. Direct comparisons of the various quinolones are too limited to date to provide clear therapeutic options. Nevertheless, this class of compounds is likely to play a major role in providing effective oral therapy for conditions that have previously required prolonged parenteral treatment.


Temafloxacin does not potentiate the anticoagulant effect of warfarin in healthy subjects. The potential for drug interaction between temafloxacin, a new fluorinated quinolone antibiotic, and low-intensity warfarin was studied in 10 healthy male volunteers. Warfarin was administered orally at titrated dosages to maintain the International Normalised Ratio between 1.3 and 1.7, and was kept constant during the last 4 days of the 16-day trial, when temafloxacin 600mg twice daily was coadministered. Prothrombin times during temafloxacin and warfarin administration were similar to those during warfarin alone. Thus, temafloxacin did not potentiate the anticoagulant effect of warfarin 1.5 to 5.5mg daily in healthy volunteers.


Pharmacokinetics of temafloxacin after multiple oral administration. Four multiple dose studies involving 102 healthy volunteers were reviewed to determine the sources of intersubject variability in the pharmacokinetics of temafloxacin. As a result of the preferential distribution of temafloxacin into muscle, liver and kidney compared with adipose tissue, the best anthropometric explanatory variable was found to be lean body mass, rather than total body mass or body surface area. Single dose studies have confirmed that the distribution volume of temafloxacin is smaller in subjects in whom the lean body mass:total body mass ratio is low, notably in females, the elderly, and those with hepatic impairment. Average creatinine clearance for the volunteers included in this analysis was normal (109 +/- 29 ml/min), and renal function was only a marginally significant covariate; however, renal clearance accounts for 60 to 70% of total clearance (CLT) and has been shown to be a major factor in the elderly, as well as in patients with renal or hepatic impairment. Since temafloxacin is partially reabsorbed renally, urine flow rate was found to be a potentially important secondary factor. Overall, the pharmacokinetics of temafloxacin are essentially linear, with steady-state plasma concentrations at trough (Cssmin) and 2h postdose (Css2h) averaging slightly more than 0.5 and 1.0 mg/L per 100mg administered every (q) 12h. For example, mean Cssmin and peak steady-state plasma concentration (Cssmax) values after administration of temafloxacin 600mg q12h were 3.3 +/- 1.1 and 6.2 +/- 1.8 mg/L, respectively. Total apparent clearance (CLT/F) and the terminal elimination half-life (t1/2) averaged 11 L/h (184 ml/min) per 55kg lean body mass and 8.4h, respectively. Considering that the data reviewed came from 4 studies with doses ranging from 100 to 800mg q12h, the intersubject coefficients of variation were low for an orally administered drug, ranging from 15.4% for t1/2, 20.6% for Css2h, 21.2% for CLT/F and 25% for Cssmin.


Pharmacokinetics of temafloxacin in patients with liver impairment. A multicentre study was conducted to determine whether liver impairment would alter the pharmacokinetics of temafloxacin, a new fluoroquinolone antimicrobial agent. 16 patients with cirrhosis and 12 healthy volunteers (the control group) received a single oral 600mg dose of temafloxacin. Blood and urine were sampled at frequent intervals after drug administration and assayed by high performance liquid chromatography. The mean age of patients with liver impairment was greater than that of the control group; they also had a lower creatinine clearance and urine output. There was no difference between the groups in either the peak plasma temafloxacin concentration or the time to reach peak concentration. However, the volume of distribution and elimination rate constant of temafloxacin were significantly lower in the group with liver impairment, as were total temafloxacin clearance, renal clearance, and the ratio of renal:creatinine clearance. Nonrenal clearance was similar in patients and controls. Creatinine clearance and urine output were found to account for most of the intersubject variability in total clearance as determined by multiple linear regression analysis. Because the altered temafloxacin pharmacokinetics appear to be primarily due to impaired renal function, this should be the main determinant of temafloxacin dosage in patients with liver disease.


Hepatobiliary elimination of temafloxacin. The biliary excretion of temafloxacin and temafloxacin glucuronide was characterised in this study after administration of a single oral temafloxacin 600mg dose to 8 patients with T-tube drainage of the common bile duct inserted after cholecystectomy or choledochotomy. High performance liquid chromatographic analyses of plasma, urine and bile samples collected during the 72h after temafloxacin administration showed that biliary concentrations of unchanged temafloxacin followed a time-course parallel to plasma concentrations but were 5- to 10-fold higher. Biliary temafloxacin peak concentrations ranged from 18.74 to 64.35 mg/L and time to peak concentrations from 0.71 to 10.23h. Mean hepatobiliary clearance of temafloxacin was 3.10 ml/min (0.19 L/h) when calculated for the unchanged drug and 1.43 ml/min (0.09 L/h) when calculated for its biliary excretion as glucuronic acid conjugates. Patients with higher bile production had markedly higher clearance of both temafloxacin and temafloxacin glucuronide. The elimination time-course of the conjugate in bile generally paralleled those of temafloxacin in bile and plasma, although there was a lag in the rate of appearance of the conjugate in bile. Biliary excretion of unchanged temafloxacin and temafloxacin glucuronide accounted for approximately 2.2 and 1% of the administered dose, respectively. Thus, it appears that hepatobiliary elimination of temafloxacin and its glucuronide acid accounts for only a small fraction of total temafloxacin clearance. Nonetheless, concentrations attained in the bile are far above the minimum inhibitory concentration values of pathogens relevant in biliary tract infections.


The relative bioavailability of temafloxacin administered through a nasogastric tube with and without enteral feeding. The relative bioavailability of a single oral dose of temafloxacin given with and without enteral feeding was determined in 18 healthy male volunteers in a randomised crossover study. Subjects were administered 600mg of temafloxacin orally as an intact tablet, or a crushed tablet suspended in water administered through a nasogastric tube with or without an enteral feeding solution [Osmolite (Ross) 100 ml/h started 2h before administration of temafloxacin and continued for 4h postdose]. Plasma samples were analysed by a high performance liquid chromatographic technique. Mean peak plasma concentrations (Cmax) for the oral tablet, crushed tablet, and crushed tablet with enteral feeding solution were 3.95 +/- 1.02, 4.85 +/- 0.69, and 4.69 +/- 0.61 mg/L/70kg, respectively, and mean calculated area under the concentration-time curve from time 0 to 48h (AUC(0-48h)) values were 48.1 +/- 11.0, 54.5 +/- 6.52, and 49.7 +/- 5.89 mg/L.h/70kg, respectively. In terms of AUC(0-48h) and Cmax, the relative bioavailability of temafloxacin after nasogastric delivery of crushed temafloxacin given with and without an enteral feeding solution was equivalent to the reference oral regimen.


The effect of food on the bioavailability of temafloxacin. A review of 3 studies. Temafloxacin is a new fluoroquinolone antibacterial agent. Previous pharmacokinetic studies have shown that the extent of absorption of temafloxacin is independent of dose and that the dispositional pharmacokinetics are linear. The pharmacokinetics of 3 tablet formulations of temafloxacin were investigated in phase I studies in healthy adult volunteers, to determine whether the bioavailability is altered by the presence of food. Results of these studies indicate that the absorption of temafloxacin is slightly enhanced by concomitant administration of food. Thus, special coordination of temafloxacin administration and meals appears to be unnecessary.


Penetration of temafloxacin into body tissues and fluids. The new fluoroquinolone antimicrobial temafloxacin shows good penetration into a range of body tissues and fluids, after single or repeated oral administration to healthy volunteers and patients undergoing various procedures. Temafloxacin concentrations in respiratory tissues and fluids, nasal secretions, tonsils, prostate, semen, bone and blister fluid were similar to, or greater than, concurrent serum concentrations. Penetration into sinus secretions and cerebrospinal fluid is less marked; nevertheless, temafloxacin concentrations of approximately 2.4 and 1 mg/L, respectively, were achieved. Concentrations of temafloxacin observed in these tissues and fluids exceed the minimum concentrations required to eradicate the majority of bacterial pathogens associated with respiratory infections, tonsillitis, sinusitis, prostatitis, bone infections and meningitis. Since elimination of temafloxacin occurs by the renal route, high concentrations of the drug are also found in the urine. Biliary excretion of temafloxacin accounts for about 3% and leads to high biliary concentrations of the drug.


Effects of 2 quinolone antibacterials, temafloxacin and enoxacin, on theophylline pharmacokinetics. Certain quinolone and naphthyridone antibacterial agents reduce the clearance of theophylline, posing potential clinical risks for patients maintained on this bronchodilator. Whether temafloxacin also affects theophylline pharmacokinetics was assessed in a randomised double-blind 3-way crossover study in 12 healthy volunteers, using placebo and enoxacin as controls. Each volunteer participated in all 3 phases of the study, receiving theophylline plus daily divided doses of temafloxacin 800mg, enoxacin 800mg, or placebo, orally for 7 days. Aminophylline 200mg (containing 146mg theophylline) was given orally twice daily on the first 4 days. On the fifth morning, theophylline 200mg was administered intravenously, and serial blood and urine samples were collected for the following 72h. Coadministration of enoxacin significantly reduced the metabolic clearance of theophylline (approximately 65%). In contrast, during coadministration of temafloxacin, theophylline pharmacokinetics did not differ significantly from those during coadministration of placebo. No clinically significant adverse events occurred; total reported adverse events during enoxacin-theophylline administration (n = 33) were higher than those reported during temafloxacin-theophylline administration (n = 22) and theophylline alone (n = 23). Administration of temafloxacin to patients on long term theophylline therapy appears to be a safe and rational choice when treatment with a broad spectrum antibiotic is indicated.


Effect of cimetidine on the pharmacokinetics of temafloxacin. Cimetidine, a widely prescribed histamine H2-receptor antagonist, is known to interact with a variety of drugs; consequently, it is important to determine its potential for interaction with any new drug. The interaction between cimetidine and a new quinolone, temafloxacin, has been examined in an open randomised 2-period crossover study in 12 healthy adults. Half the volunteers received cimetidine 400mg 3 times daily for 8 days. On day 5, a single temafloxacin 400mg dose was administered. No other drugs were allowed during this period. The other volunteers did not receive cimetidine (or any other drugs) for an 8-day period except for temafloxacin 400mg on day 5. Blood and urine were sampled serially after temafloxacin administration and daily thereafter until the last day of the study. A 2-week washout period preceded crossover. Pharmacokinetic analyses showed that cimetidine did not affect the rate or extent of temafloxacin absorption, as evidenced by unchanged peak plasma concentration, time to peak plasma concentration, and terminal-phase volume of distribution. However, renal and total clearance values for temafloxacin were reduced by 19%, and elimination half-life and area under the concentration vs time curve were increased in the presence of cimetidine. The most likely mechanism underlying these effects is inhibition by cimetidine of tubular secretion of temafloxacin in the kidney. The lack of clinically significant adverse effects and the small magnitude of the reduction in temafloxacin clearance suggest that the interaction is of little clinical consequence.


Effect of antacid medication on the pharmacokinetics of temafloxacin. The effect of an antacid drug (Maalox 70) on the pharmacokinetics of temafloxacin was studied in 12 healthy young volunteers. The study was designed as a randomised open 2-period crossover trial in which temafloxacin was administered alone and with Maalox 70. In both treatments, temafloxacin was administered as a single oral 400mg dose on the morning of day 2. In the antacid regimen, 8 doses of Maalox 70 were administered every 2h on day 1, starting at 8am and ending with the last dose at 10pm; 5 doses were given on day 2, at 2.5 and 1h before administration of temafloxacin and 1, 3 and 5h after the temafloxacin dose. With coadministration of Maalox 70, peak plasma temafloxacin concentrations (Cmax) were reduced to 44.6% (+/- 24.5), and AUC(0-infinity) was reduced to 39.8% (+/- 17.4) of the corresponding values obtained when temafloxacin was given alone. Urinary excretion of temafloxacin was reduced to 46.0% (+/- 13.7) of that observed when temafloxacin was administered alone. Time to peak plasma concentration (tmax = 1.8h) was not affected by antacid administration. Comparable or greater antacid-associated reductions in relative bioavailability have been reported for other quinolones. As with other quinolones, the concurrent administration of temafloxacin and antacids should be avoided.


Effects of temafloxacin and ciprofloxacin on the pharmacokinetics of caffeine. A number of quinolone antibacterial agents, particularly enoxacin, pefloxacin, pipemidic acid and ciprofloxacin, are known to decrease the clearance of methylxanthines. The effects of temafloxacin and ciprofloxacin on the pharmacokinetics of caffeine were therefore compared in a 3-way crossover study in 12 healthy young volunteers. Each volunteer received 183mg once-daily doses of caffeine in conjunction with twice-daily placebo, temafloxacin 600mg and ciprofloxacin 750mg in 3 separate phases according to a randomised sequence. A doubling of the area under the plasma concentration-time curve (77.8 vs 31.8 mg/L.h) and terminal-phase half-life (9.7 vs 4.5h) of caffeine were observed in the presence of ciprofloxacin. The magnitude of the reduction in the intrinsic clearance of caffeine produced by ciprofloxacin was greater than that described in the literature for ciprofloxacin and theophylline. This may partly be explained by intertrial differences in dosage and study design. Coadministration of temafloxacin did not have any effect on the pharmacokinetics of caffeine, confirming results of other studies suggesting that this agent does not affect methylxanthine clearance. Accordingly, it appears that restriction of caffeine intake during temafloxacin therapy is not necessary.


Absence of drug interaction between temafloxacin and low dose heparin. An open 5-day study was conducted in 12 healthy male volunteers to determine the potential for drug interaction between low dose standard heparin and temafloxacin, a new fluorinated quinolone antibiotic. Heparin 5000IU was administered subcutaneously on the morning of the first and last study days and temafloxacin 600mg was administered twice daily for 4 days. The mean change in activated factor X (antifactor Xa) concentration relative to baseline 2h after coadministration was similar to that after heparin alone. Likewise, changes in activated partial prothrombin time, prothrombin time and thrombin time were similar after either heparin alone or concomitant temafloxacin administration, indicating the absence of interaction between temafloxacin and low dose standard heparin in healthy volunteers.


Lack of correlation between anticardiolipin antibodies and peripheral autonomic nerve involvement in systemic lupus erythematosus. The presence of anticardiolipin antibodies has recently been related to a clinical complex in which both central and peripheral neurologic damage is included. A series of 27 female patients affected by systemic lupus erythematosus (SLE) was tested for the presence of peripheral autonomic neuropathy and serum anticardiolipin antibody (ACA) levels were determined in each patient by ELISA. Peripheral autonomic impairment was detected in 40.7% of SLE patients and a large number (77.7%) of patients had elevated levels of ACA. No relationship was found between presence of ACA (both for IgG and IgM classes) and the autonomic neuropathy.


Inhibition of superoxide anion release from human polymorphonuclear leukocytes by N-acetyl-galactosamine and N-acetyl-glucosamine. This study shows that N-acetyl-galactosamine and N-acetyl-glucosamine can diminish the production of superoxide anion from cytochalasin-B treated PMNs stimulated with FMLP. Inhibition ranged from 80.9% to 1.8%. N-acetyl-galactosamine was superior to N-acetyl-glucosamine, but both showed their action in a dose-related fashion. The mannosamine may diminish the superoxide production, but without a statistical significance. Other sugars such as L-fucose, D-fucose and D-glucose failed to induce inhibition of superoxide generation. Previous reports showed that sugars interfere with carbohydrates lectins interaction. This study shows that aminosugars can do more than interfere with carbohydrates-lectin interaction. The mechanism is not completely known yet. The question whether aminosugars affect cell-cell interaction, regulation of respiratory burst, inflammatory mediators functions, or the glucose uptake and utilization needs further study.


Treatment of pigmented villonodular synovitis with yttrium-90: changes in immunologic features, Tc-99m uptake measurements, and MR imaging of one case. Pigmented villonodular synovitis (PVNS) is an uncommon proliferative disease of synovium. We report a 35-year-old male with diffuse form of PVNS of left knee, treated with intraarticular injection of 5 mCi of yttrium-90 (Y-90) silicate colloid consisting of two doses with a 3-month interval between them. During follow-up, the affected knee showed clinical improvement and was accompanied by a decrease of the levels of soluble interleukin-2 receptor in sera and synovial fluids (SF). When compared to osteoarthritis subjects, SF lymphocyte subsets of this case before Y-90 therapy showed a lower CD4:CD8 cell ratio and absence of suppressor inducer cells (CD4+ 2H4+). The Tc-99m pertechnetate knee uptake indexes correlated well with clinical improvement. Serial magnetic resonance imaging revealed significant change one year after Y-90 therapy. The findings of immunological assessment suggested that immunoregulatory dysfunction may be related to the pathogenesis of PVNS.


Nitrite biotransformation by mitochondria from the earthworm Eisenia foetida (Savigny). 1. Nitrite oxidase and cytochrome-c oxidase activity catalysed by cytochrome-aa3 were assayed in earthworms and rats. 2. Cytochrome-aa3 and intact mitochondria from the two species were anaerobically incubated in the presence of nitrite; the occurrence of mitochondria-induced nitrite biotransformations was evaluated by monitoring nitrite recovery in incubation medium. Possible nitric oxide production was also tested. 3. The ratio nitrite oxidase/cytochrome-c oxidase activity was much higher in earthworms than in rats. 4. Under anaerobic conditions and in the presence of respiratory substrates, earthworm mitochondria produced a time-dependent loss of nitrite in the incubation medium. On the contrary, rat mitochondria are unable to decrease environmental nitrite concentration. 5. Results support the notion that metabolic properties of earthworm mitochondria can be considered as an adaptation to chronic nitrite exposure, this toxicant being typically present in natural habitats of these worms.


The role of flow cytometry and morphometry in discriminating benign from malignant nonfunctioning pancreatic endocrine tumors. Nonfunctioning pancreatic endocrine tumor (NFPET) is a kind of tumor with high malignancy but bland in morphological features. It is impossible to predict the malignancy solely by gross and light microscopic pictures of the tumor. For the purpose of assessing the value of flow cytometry and morphometry in predicting the prognosis of NFPET, the DNA content and several morphological parameters of 26 NFPETs were determined by appropriate techniques. The results show that abnormal DNA content has no practical value in discriminating benign from malignant NFPETs; whereas the "score" obtained by combining the morphological measurements of the number of nuclei/mm2 and N/C, is of certain referential significance in predicting the malignancy of NFPET.


Detection of human papillomavirus DNA sequences in cervical lesions by in situ hybridization using biotinylated DNA probes. Ninety-six biopsy specimens were collected for studying the association of specific sequences of HPV DNA with genital warts, precancerous lesions and invasive cervical cancer. Using DNA probes labeled with biotin, 17 of the 22 (77.27%) specimens showing condyloma and 1 of the 14 (7.14%) specimens displaying subclinical papillomavirus infection (SPI) of cervix were found to be positive for HPV 6/11; 4 of 23 (17.39%) specimens of squamous epithelial cervical cancer and 2 of the 23 (8.70%) specimens of cervical intraepithelial neoplasia (CIN) were positive for HPV 16/18. All the 14 specimens of cervicitis were negative for both probes. The HPV 6/11 positive hybridization products were present mainly in the superficial and intermediate epithelium of condyloma and SPI. In invasive cancer, HPV 16/18 were shown more often in cancer cells of infiltrative nests and were focally distributed.


[Surgical treatment of primary small cell carcinoma of the esophagus. Clinical analysis of 11 cases]. 545 cases of esophageal carcinoma were confirmed histologically and by esophagoscopic biopsy between Jan. 1982 and May 1990. Primary small cell carcinoma was identified in 19 cases. Of 11 patients confirmed operatively, 5 had oat cell carcinoma, 4 combined oat cell carcinoma and 2 intermediate cell carcinoma. The mean overall survival period was 13.9 months. The longest survival period was 27 months. The prognosis of primary small cell carcinoma was poorer than that of squamous carcinoma of the esophagus because of its propensity of spread and metastasis. Once the diagnosis of small cell carcinoma was established, surgery was undertaken as early as possible. For patients who had lymph node metastasis at the time of diagnosis, we recommended surgical treatment plus systemic chemotherapy after operation. To increase the resection rate, it is important to do chest CT scan, bronchiofiberscopy and B-type ultrasonography before operation.


Antiulcer drugs and gastric mucosal integrity. Effects of misoprostol, 16,16-dimethyl PGE2, and cimetidine on hemodynamics and metabolic rate in canine gastric mucosa. Although prostaglandins (PGs) of the E series have gastric antisecretory and cytoprotective properties, many have different effects on the barrier integrity of the gastric mucosa. The direct effect of antiulcer drugs on gastric mucosal blood flow, mucosal barrier permeability, and metabolic rate have not been adequately studied. These factors are important in the defense of the gastric mucosa. Part of the difficulty relates to the possible influence of gastric mucosal blood flow on gastric acid secretion. To rule out this confounding factor, omeprazole can be used to reveal the true pharmacologic effects of these antiulcer drugs independent of the effect of gastric secretion per se. The study examined the effects of 16,16-dimethyl PGE2 (dmPGE2), misoprostol, and cimetidine on gastric mucosal blood flow, oxygen consumption, potential difference (PD), electrolytes, and fluid flux using the ex vivo gastric chamber dog model. The PGs were administered intraluminally with an isotonic acid solution; cimetidine was administered by arterial infusion. None of the drug treatments had any significant effect on mean systemic arterial pressure, arterial blood gases, body temperature, or oxygen consumption. dmPGE2 significantly (P less than 0.001) decreased PD and enhanced the electrolytes (Na+, K+) and fluid flux across the mucosa (P less than 0.05). Misoprostol significantly increased gastric mucosal blood flow (P less than 0.02) and fluid efflux but decreased PD values. Cimetidine did not have any significant effects on barrier or metabolic functions of the stomach. These results suggest that a considerable difference exists in the pharmacology of gastric antisecretory drugs in relation to their effect on several factors affecting gastric mucosal integrity.


Prevalence of hepatitis C virus antibody in a liver transplantation population. The development of a serologic assay to detect antibodies directed at an antigen (C-100-3) of the hepatitis C virus (anti-HCV) has been a major breakthrough in the long search for causative agents of non-A, non-B (NANB) hepatitis. The frequency of HCV in those who have end-stage liver disease is not known. Moreover, the rate of recurrence after liver transplantation (OLTx) and the rate of acquisition of new HCV infection as a result of the OLTx experience is as yet unknown. This study was performed in an attempt to answer these questions. The prevalence of HCV in 372 patients undergoing OLTx at the University of Pittsburgh was determined. Those transplanted for HBV-related liver disease with hepatoma had the highest rate of HCV antibody positivity (45.4%) followed by those with metabolic liver disease (42.5%), putative NANB liver disease (41.4%), and cryptogenic cirrhosis (20.9%); those with cholestatic liver disease exhibited the lowest rate (16.2%). HCV antibody was positive in only 26.3% of patients with hepatoma. Of those patients who were negative prior to transplantation, 12.2% acquired HCV antibody post-OLTx. In the putative NANB group, no difference was detected in the AST and ALT prior to transplantation in either the HCV antibody-positive or -negative patients. In patients with cryptogenic cirrhosis, those who were positive for HCV antibody had higher transaminase levels prior to transplantation than did those patients who were HCV antibody negative.


[Nasopharyngeal carcinoma (NPC) mortality rate in two districts of Guangdong Province and their relation to IgA antibody level against EB virus capsid antigen]. Based upon the retrospective investigation data, the NPC mortality rate in the Pearl River Delta was found to be twice as high as that of Chaoshan district. The positive rate of IgA/VCA antibody in the Pearl River Delta was also found to be significantly higher than that in the Chaoshan district by viro-serological screening. The IgA/VCA antibody positive rate and mortality rate were concomitantly increased with age in the bracket of 30-59. Accordingly, the authors assume that activation of infected EB virus might be an important factor in the process of etiology-pathogenesis of NPC.


[Factors influencing recurrence and metastasis after curative resection for rectal carcinoma]. This paper analyzes 737 cases of curative resection for rectal carcinoma treated from January 1958 to December 1987. Males 407 and females 330. The ages ranged from 17-84. The follow up period was 1 to 30 years. 217 patients developed recurrence or metastasis and the disease-free survival was 3 months to 15.5 years with a median of 24.2 months. Most of the recurrence occurred in 2 years and was found in the pelvis after abdominoperineal resection and the site of anastomosis after anterior resection. The chief metastatic sites were: lung, liver and inguinal nodes in order of frequency. Factors influencing recurrence and metastasis were: site of cancer, Dukes stage, pathologic type, differentiation of cancer, node involvement, and method of operation. Adjuvant irradiation could reduce local recurrence. Patients prone to developing recurrence should be closely followed and the resectable lesions be resected whenever possible.


[Value of serum glycosidase spectrum in the diagnosis of hepatocellular carcinoma]. Activities of alpha-L-fucosidase (alpha-Fucase), N-acetyl-beta-D-glucosaminidase (beta-GlcNA-case), N-acetyl-beta-D-galactosaminidase (beta-GalNAcase) and alpha-mannosidase (alpha-Manase) in sera of normal adults, patients with hepatocellular carcinoma (HCC), benign liver diseases and non-liver diseases were determined by microquantitative spectrophotometry. The results showed that the activity of the four serum glycosidases in patients with HCC was significantly higher than that in normal adults. When the maximum 95% confidence limit was used as the positive line, the positivities of alpha-Fucase, beta-GlcNAcase, beta-GalNAcase and alpha-Manase for the diagnosis of HCC were 66.80%, 37.29%, 32.20% and 18.64%, respectively. There was a close relationship among the four glycosidases without being related to serum alpha-fetoprotein (AFP). Some patients with benign liver diseases and non-liver diseases also had elevated glycosidase activity. However, the increase in several glycosidase activities was mainly found in HCC patients. Hepatitis patients with increased activities of more than one glycosidase were always accompanied with elevated SGPT or other abnormal liver functions. Therefore, serum glycosidase spectrum is useful for the diagnosis and differential diagnosis of HCC, especially in AFP negative HCC patients.


[Surgical treatment of primary liver cancer complicated with cirrhosis]. From 1964 to 1990, 318 cases of primary liver cancer were treated by surgery. Of them, 227 (71.3%) were complicated with cirrhosis to various degrees: mild, moderate and severe. The lesions in the latter group was greater than 10 cm in 3 patients, 5-10 cm in 7 and less than 5 cm in 17. All were confirmed by histopathology to be hepatocellular carcinoma. The method of operation was partial hepatectomy in 23 patients and intra-tumoral absolate alcohol injection supplemented by microwave coagulation in 4. The amount of absolate alcohol ranged from 18 to 30 ml with an average of 24 ml. The chief complications were jaundice and ascites (13 patients--48%) and gastrointestinal hemorrhage (3 patients--11%). Operative mortality was 15% (4/27). The 1-, 3- and 5-year survival rates were 82%, 39% and 10%.


[Expression and significance of Pre-S1 and Pre-S2 in human primary hepatic carcinoma (PHC)]. 135 specimens of primary hepatic carcinoma (PHC) were formalin fixed and paraffin embedded and stained for Pre-S1, Pre-S2 and HBxAg by ABC method, for HBsAg and HBcAg by PAP method. The detection rates of Pre-S1 and Pre-S2 positive cases in cancerous tissues were 22.2% and 20.0%. The detection rates of Pre-S1 and Pre-S2 in non-cancerous liver tissues were 60.0% and 59.6%. The positive ratio of Pre-S1 and Pre-S2 in the same hepatoma was 16.3% and that in the same non-cancerous liver tissue was 55.6%. Among 135 cases of PHC, HBsAg, HBxAg and HBcAg positives in tumor tissues were 16.3%, 55.6% and 8.9%, respectively. Those in non-cancerous tissues were 59.6%, 78.8% and 24.2%. This study suggested that the detection rates of Pre-S1 and Pre-S2 positivity in hepatoma tissues were higher than those of HBsAg and HBcAg but lower than that of HBxAg. The frequency of positive Pre-S1 and Pre-S2 in non-cancerous liver tissues was similar to HBsAg, and slightly lower than that of HBxAg. S1 and S2 are considered new markers for HBV infection. Their antigens could play an important role in the pathogenesis of PHC.


A quantitative and interspecific test for biological activity of anti-müllerian hormone: the fetal ovary aromatase assay. Anti-Müllerian hormone (AMH), also known as Müllerian-inhibiting substance or factor, has previously been shown to sex-reverse the steroidogenic pattern of fetal mammalian ovaries through repression of aromatase biosynthesis. Study of the ontogeny of the response of cyclic AMP-stimulated aromatase activity of rat fetal ovaries to AMH has allowed us to develop a quantitative bioassay for the hormone. Linear responses as a function of the logarithm of AMH concentration were observed over ranges of 0.2-7.5 micrograms/ml for the bovine protein and 0.15-2 micrograms/ml for the human protein, with a maximal decrease in aromatase activity of 90% for both proteins. Under the same in vitro conditions, AMH treatment did not affect cyclic AMP-stimulated fetal rat testicular aromatase activity. Partially purified chick AMH also decreased rat ovarian aromatase activity, allowing us to use this test to study AMH ontogeny in chick gonads. Analysis of the species specificity of AMH repression of ovarian aromatase activity indicated that turtle and rat fetal ovaries responded to AMH of other vertebrate classes, whereas aromatase activity of chick embryo ovaries could be repressed only by the homospecific hormone.


Development of the spatial pattern of retinoic acid receptor-beta transcripts in embryonic chick facial primordia. Retinoic acid causes a range of embryonic defects, including craniofacial abnormalities, in both birds and mammals and is believed to have a number of roles in normal development. We have previously shown that the distribution of retinoic acid receptor-beta (RAR-beta) transcripts is spatially restricted within the neural-crest-derived upper beak primordia of the chick embryo. We have now used in situ hybridisation to trace the distribution of RAR-beta transcripts during the migration of cranial neural crest cells and during formation of these primordia. RAR-beta transcripts were present in a subset of migrating neural-crest-derived cells in the head of the stage 10 embryo. These cells were situated in pathways followed by cells that migrate from the neural crest overlying the posterior prosencephalic/anterior mesencephalic region of the developing brain. Cells containing RAR-beta transcripts accumulated around the developing eyes and in the regions of the ventral head from which the upper beak primordia later develop. We mapped the distribution of RAR-beta transcripts as the facial primordia were forming, with particular reference to the development of the maxillary primordia. We found that these form in a region of the ventral head that includes the boundary between regions of high and low levels of RAR-beta transcripts. The boundary between these two groups of cells persisted as the maxillary primordia developed.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of the two mannose 6-phosphate receptors is spatially and temporally different during mouse embryogenesis. Mammalian cells express two mannose 6-phosphate receptors, MPR46 and MPR300, both of which mediate the targeting of lysosomal enzymes to lysosomes. Additionally the receptors mediate the secretion (MPR46) and the endocytosis (MPR300) of lysosomal enzymes and the binding of IGFII (MPR300). We have analyzed the distribution of MPR46 and MPR300 transcripts during mouse embryogenesis by in situ hybridization. Up to day 15.5 of embryonic development we found a non-overlapping distribution of the transcripts for the two receptors. High expression of MPR46 was observed at sites of hemopoiesis and in the thymus while MPR300 was highly expressed in the cardiovascular system. Late in embryogenesis (day 17.5) a wide variety of tissues expressed the receptors, but still the expression pattern was almost non-overlapping. This unexpected complementary expression pattern points to specific functions of the two mannose 6-phosphate receptors during mouse embryogenesis.


Loss of type II calcium/calmodulin-dependent kinase activity correlates with stages of development of electrographic seizures in status epilepticus in rat. Understanding the molecular basis of altered neuronal excitability in epilepsy is a major challenge in neuroscience research. The present study suggests an inverse correlation between changes in neuronal excitability in status epilepticus and the activity of type II multifunctional calcium/calmodulin-dependent kinase II (CaM kinase II), a major Ca(2+)-signal transducing system in brain. 'Continuous' hippocampal stimulation (CHS), a new model of non-convulsive limbic status epilepticus (SE), mimics the progression of electrographic changes characteristic in human SE and allows for quantitation of post-stimulus seizure severity. In the present study, hippocampus and anterior neocortex from CHS-stimulated rats and paired surgical controls were assayed for CaM kinase II activity by incorporation of radiolabeled phosphate from [gamma-32P]ATP into the 50-kDa subunit of the kinase itself (autophosphorylation). In all instances, CHS induced sustained interictal bursting and/or electrographic seizures. Decreased CaM kinase II activity was seen in all preparations from electrically stimulated hippocampus. CaM kinase II activity in CHS animals was diminished by 37% relative to controls (P less than 0.01; Student's paired t-test). The progressive intensity of the EEG discharges correlated directly with the decrement of CaM kinase II activity (P less than 0.05; Spearman's rank correlation test, n = 5). This is the first report of a dynamic modulation of a biochemical system that has been implicated in neuronal excitability in coordination with the characterized developmental stages of SE.


Chronic liver disease in Ethiopia: a clinical study with emphasis on identifying common causes. Between July 1986 and April 1989, 334 hospitalized adult Ethiopian patients with chronic liver disease were studied according to a protocol to define their clinical features and to identify risk factors with the aim of preventive intervention. Of these, 14 had chronic hepatitis, 208 cirrhosis and 112 hepatocellular carcinoma (HCC). Both clinical and histological diagnostic criteria were employed. A detailed questionnaire was used to document demographic and clinical data. A common clinical presentation among patients with chronic hepatitis was darkening of the face and hands with or without hypertrichosis of the face and blisters over the dorsi of the hands. This overt or latent form of porphyrea cutanea tarda (PCT) responds to chloroquine. Patients with cirrhosis of the liver commonly present for the first time with ascites, splenomegaly, haematemesis and/or melena from oesophageal varices, and mental changes due to hepatic encephalopathy. Overt or latent forms of PCT are also common features. Peculiar to these cirrhotics is the rarity of spider naevi, gynaecomastia, testicular atrophy, Dupuytren's contracture, parotid gland enlargement and clubbing of the fingers. Exhaustion, loss of appetite, rapid loss of weight, right upper quadrant and/or epigastric pain (all often of less than 6 months' duration, a big, hard, tender and grossly nodular liver with bruit, signs of portal hypertension, and/or hepatic encephalopathy, in a young male with a rapid down hill course characterize the Ethiopian patient with HCC. Serum anti-nuclear factor, anti-mitochondrial anti-bodies and anti-smooth muscle anti-bodies were absent in those with chronic hepatitis and were uncommon in the cirrhotics and HCC cases. One or more hepatitis B virus markers were found in 86% of chronic hepatitis, 88% cirrhosis and 78% HCC and the HBsAg carrier state was found in 36%, 29% and 23%, respectively. Among the HBsAg carriers, HBeAg positivity was less common than anti-HBe but anti-HDV was significantly higher than in the healthy general population. Alphafetoprotein (AFP) levels greater than 500 mg/ml were present in 16 (8%) cirrhotics and 58 (52%) patients with HCC. Histologically, 3 of the chronic hepatitis patients had progressed to cirrhosis, 8 of the cirrhotic patients had chronic active hepatitis and 85% of HCC cases occurred in a background of macronodular cirrhosis. Three cirrhotics developed HCC during follow-up.(ABSTRACT TRUNCATED AT 400 WORDS)


Histological human papillomavirus induced lesions: typization by molecular hybridization techniques. The viral typization in Human Papillomavirus (HPV) infections of the lower female genital tract is relevant both from the epidemiological and the clinical point of view. We have tested DNA from specimens obtained by guided biopsies on neoplastic and benign lesions, using single probes of the different virus types (6, 11, 16 and 18) by Dot-blot and Southern blot analysis. According to previous studies HPV 16 and 18 have been detected in neoplastic lesions, while 6 and 11 were more frequently found associated to condylomata. Negative specimens and intermediate were 36% of the total. In our experience both methods have shown same sensibility.


Human papillomavirus infection of the female genital tract: correlation of HPV DNA with cytologic, colposcopic, and natural history findings. Although molecular biology evidence suggests a strong link between HPVs and anogenital neoplasias, evidence from clinical studies is still less convincing. Thus, the recognition of both overt and subclinical HPV infection has become increasingly important. We studied the correlation between HPV DNA and cytologic, colposcopic and natural history findings in a defined group of women. The study population consisted of 150 HPV DNA positive cases (mean age 25.7 years, SD 7.4) and 69 randomly selected HPV DNA negative controls (mean age 27.3 years, SD 7.9) enrolled in an ongoing study of the natural history of genital HPV infections. All cases and controls had normal cervicovaginal cytology at the enrollment. A commercial dot-blot technique hybridization test (Virapap and Viratype, Digene Diagnostics, USA) was used for HPV DNA testing of cervicovaginal scrapes. Five percent of the patients had HPV 16/18, 31% had HPV 31/33/35, 15% had more than one of the three HPV DNA groups ("mixed"), and 15% had untypable HPV DNA. Cases and controls were followed by repeat examinations every 4th month. The mean follow-up time was 12.2 months (SD 8.7) for the cases, and 12.8 months (SD 6.9) for the controls. The study endpoint was defined as the presence of cytologic changes consistent with CIN. The overall prevalence of atypical transformation zone (ATZ) findings was 45% in the controls and 56% in the cases, with no significant differences between the specific HPV DNA groups. Although vulvar or vaginal abnormalities (acetowhite epithelium, squamous papillomatosis, filaments, satellite lesions, fissures, papules or exophytic condylomas) were more commonly seen in the cases than in the controls, the difference was significant only for condylomas and fissures.(ABSTRACT TRUNCATED AT 250 WORDS)


Nucleolar organizing regions in human papillomavirus infection and in cervical intraepithelial neoplasia. The Ag-NOR staining technique for nucleolar organizer regions (Ag-NORs) was applied to 37 biopsies of the uterine cervix taken from colposcopically abnormal areas. Histological examination of the sections showed that 13 patients had condylomatous lesions without atypia, 15 had CIN I, 4 had CIN II, 3 had CIN III and 2 carcinoma in situ. The mean numbers of Ag-NORs in parabasal and basal cells of squamous epithelium increased progressively from CIN I to CIN III-CIS. There was no significant difference between the Ag-NOR count in condylomatous lesions and CIN I. Significant differences were found between the number of Ag-NORs in condylomatous lesions and in the various grades of CIN. The Ag-NOR technique is useful in the diagnosis of cervical intraepithelial neoplasia and confirms the need to regard condylomata of the uterine cervix as intraepithelial neoplasia grade I and recommend these patients for follow-up.


Dynamic study of methionine uptake in glioma using positron emission tomography. In order to evaluate the methionine uptake of a glioma with positron emission tomography (PET), the kinetics of carbon-11 methionine was investigated in 11 patients by measuring the free 11C-methionine in plasma as an input function following intravenous administration. When the mean clearance curve of free 11C-methionine in plasma instead of individual 11C-methionine clearance curves was used, mean differences in uptake rate and distribution volume were 4.0% and 4.6% respectively. By applying the mean clearance curve of free 11C-methionine in 18 glioma patients, significant differences in 11C-methionine uptake rate and distribution volume were found according to pathological grading. For the accurate evaluation of the metabolism of 11C-methionine, it is therefore preferable that the actual level of free 11C-methionine in the plasma be measured, especially for the follow-up of individual cases. The study also demonstrated that the mean clearance curve of 11C-methionine in plasma might be employed as an input curve for calculating the uptake rate and distribution volume with small errors.


5-Hydroxytryptamine stimulates cyclic AMP formation in the tunica muscularis mucosae of the rat oesophagus via 5-HT4 receptors. The nature of 5-HT4 receptor coupling in the tunica muscularis mucosae of the rat oesophagus has been studied. 5-HT and renzapride stimulated cyclic AMP formation concentration dependently, with -log EC50 values of 7.1 and 6.8, respectively. Renzapride, relative to 5-HT, acted as a partial agonist. Tropisetron (ICS 205 930) and a novel 5-HT4 antagonist, SDZ 205 557, inhibited 5-HT-induced cyclic AMP production competitively, with pA2 estimates of 6.7 and 7.7, respectively. These data are consistent with the hypothesis that 5-HT4 receptors mediate relaxation of the smooth muscle cells of the tunica muscularis mucosae of rat oesophagus via activation of adenylyl cyclase.


Sumatriptan and 5-benzyloxytryptamine: contractility of two 5-HT1D receptor ligands in canine saphenous veins. Sumatriptan and 5-benzyloxytryptamine are ligands with high affinity for 5-HT1D receptors in the caudate nucleus. Both compounds contracted canine saphenous veins, in vitro. Benzyloxytryptamine was less potent as a contractile agonist than sumatriptan which was less potent than serotonin. In high concentrations (greater than 10(-5) M) serotonin-induced contraction resulted, in part, from activation of alpha-adrenoceptors as determined by blockade of contraction with prazosin (10(-6) M) and idazoxan (10(-6) M). Likewise, benzyloxytryptamine but not sumatriptan also activated contractile alpha-receptors in the canine saphenous vein. Furthermore, benzyloxytryptamine antagonized contraction to sumatriptan in an apparently non-competitive fashion. Thus, benzyloxytryptamine, although possessing some alpha-receptor agonist activity, like sumatriptan, can interact with serotonin receptors in canine saphenous veins. Although effects of sumatriptan and benzyloxytryptamine quantitatively differed in canine saphenous veins, both agents showed similar affinity and agonist efficacy at 5-HT1D receptors in brain. These studies may reflect potential differences between the 5-HT1D receptor in brain and the 5-HT1-like receptor in canine saphenous veins.


Effects of proton pump inhibitor on gastric mucosa hemodynamics and tissue oxygenation in anesthetized rats. Proton pump inhibitors have been reported to have a cytoprotective action in addition to the anti-secretory action of acid. The precise mechanism, however, remains obscure. In this study, the effects of proton pump inhibitors (omeprazole and NC-1300) on gastric mucosa hemodynamics and tissue oxygenation were investigated using organ reflectance spectrophotometry in a hemorrhagic shock-reperfusion model involving anesthetized rats. Neither drug affected gastric mucosa hemodynamics nor tissue oxygenation in the basal state before hemorrhage. During the hemorrhagic shock state, however, these drugs maintained tissue oxygenation and reduced ulcer formation, although they did not show a significant effect on gastric mucosa blood volume. The results suggest that both proton pump inhibitors have an anti-ulcer action by maintaining mucosal oxygenation in addition to the anti-secretory activity of acid.


Facilitation of brain stimulation reward by mesencephalic injections of neurotensin-(1-13). The effects on brain stimulation reward of neurotensin-(1-13) microinjected at different concentrations (2.5, 5, 10 and 20 micrograms/0.5 microliters) into the ventral mesencephalic region containing mesocorticolimbic dopamine neurons were tested in 12 male rats. Neurotensin lowered the stimulation frequency required to sustain threshold levels of responding for brain stimulation reward, suggesting that this neuropeptide is involved in modulating the activity of dopamine neurons that mediate behaviors motivated by positive reinforces. The magnitude of the facilitatory effect of neurotensin on brain stimulation reward was dependent on the concentration injected and to a significant extent also on whether the peptide was administered in an ascending or a descending order of concentration. The different effects of neurotensin depending on the order of administration may suggest long-lasting effects on the responsiveness of neurotensin receptors in this region after injection of high concentrations of the peptide. Subsequent injection of morphine (2.5-5 micrograms/0.5 microliter) into the same site produced a weaker facilitation of brain stimulation reward than expected, suggesting that local damage after multiple central injections or prior injections of neurotensin itself reduced the responsiveness of dopamine neurons to opiates. Taken together, the results are consistent with data indicating that activation of neurotensin receptors in the ventral mesencephalon stimulates dopamine cell firing and axonal dopamine release in limbic terminal fields and suggest that endogenous neurotensin is involved in the control of behavior motivated by positive reinforcement.


Mechanisms underlying the inhibitory effect of dibutyryl cyclic AMP in vascular smooth muscle. The mechanism by which dibutyryl cyclic AMP (db-cAMP) induces vasodilatation was examined in isolated rat aorta. The contraction induced by norepinephrine (NE) was more sensitive to the inhibitory effect of db-cAMP than that induced by high K+, and the contraction induced by lower concentrations of each stimulant was more sensitive to db-cAMP than that induced by higher concentrations. Db-cAMP at 10 microM inhibited the increases in muscle tension and cytosolic Ca2+ level ([Ca2+]i) without changing the [Ca2+]i-tension relationship, suggesting that the inhibitory effect is mainly due to a decrease in [Ca2+]i. A higher concentration (300 microM) of db-cAMP inhibited muscle tension more strongly than [Ca2+]i suggesting that db-cAMP decreases Ca2+ sensitivity of contractile elements. In contrast, 10 microM verapamil inhibited the NE-stimulated [Ca2+]i more strongly than the NE-induced contraction. The verapamil-insensitive portion of the NE-stimulated [Ca2+]i and contraction was inhibited by db-cAMP, suggesting that db-cAMP and verapamil act by different mechanisms. In Ca(2+)-free solution, 1 microM NE induced transient increases in muscle tension and [Ca2+]i. The transient contraction was inhibited by 1 mM db-cAMP more strongly than [Ca2+]i. An activator of adenylate cyclase, forskolin, showed inhibitory effects similar to those of db-cAMP. The inhibitory effects of db-cAMP and forskolin were inversely proportional to [Ca2+]i before the addition of these inhibitors. These results suggest that db-cAMP inhibits smooth muscle contraction by decreasing [Ca2+]i and the Ca2+ sensitivity of contractile elements, and that both of these effects are stronger when [Ca2+]i is lower.


The actions of 3-aminopropanephosphinic acid at GABAB receptors in rat hippocampus. The actions of 3-aminopropanephosphinic acid (APPA) were examined using whole-cell patch-clamp recording in rat hippocampal slice. In recordings from neurons in subfield CA1 of slices from young (2-4 weeks) and adult (greater than 2 month) rats, APPA (0.5-50 microM) produced membrane hyperpolarization and outward current under voltage-clamp. APPA also inhibited excitatory postsynaptic potentials with an IC50 of 2.3 microM, and reduced inhibitory postsynaptic potentials at concentrations from 0.1 to 1 microM. The hyperpolarizing and synaptic depressant effects of APPA were reduced by 2-OH-saclofen an antagonist at the B-type receptor for the neurotransmitter gamma-aminobutyric acid (GABA). In this preparation APPA exhibited potencies similar to those previously reported for the GABAB receptor agonist baclofen. APPA was much less effective in inhibiting synaptic transmission measured using field potential recordings. The observations made with whole-cell patch-clamp recording indicate that in hippocampus APPA acts as a potent agonist at presynaptic GABAB receptors associated with both excitatory and inhibitory synapses, and also activates postsynaptic GABAB receptors.


L-alpha-glycerylphosphorylcholine antagonizes scopolamine-induced amnesia and enhances hippocampal cholinergic transmission in the rat. The effects of L-alpha-glycerylphosphorylcholine (alpha-GPC) on scopolamine-induced memory impairment and on brain acetylcholine (ACh) synthesis and release were investigated in rats. Oral administration of alpha-GPC 3 h before the behavioural test prevented the learning impairment induced by scopolamine given 30 min before the acquisition of a passive avoidance response. Similarly, retrograde amnesia induced by scopolamine, given immediately after acquisition training, was also completely reversed by the drug. These effects were dose-dependent with a maximum at 300 mg/kg. The mechanism of action of this compound was investigated by measuring hippocampal ACh synthesis and release both in vivo by means of the microdialysis technique and in vitro in tissue slices. alpha-GPC dose dependently increased ACh release with a maximum at 300 mg/kg. In addition, i.v. injection of [14C]alpha-GPC resulted in [14C]ACh formation. The data suggest that the behavioural effects of alpha-GPC may be related to its property to increase hippocampal ACh synthesis and release.


Potentiation of opioid analgesia by the antidepressant nefazodone. Nefazodone is a new antidepressant related structurally to trazodone. In addition to its activity in preclinical assays for antidepressant activity, nefazodone was a potent analgesic in the mouse hotplate assay. At 50 mg/kg s.c. nefazodone doubled baseline latencies in 40% of mice but was inactive in the tailflick test at any dose tested. The hotplate analgesia seen with nefazodone alone was not reversed by naloxone (10 mg/kg s.c.). In the tailflick assay, nefazodone (50 mg/kg s.c.) enhanced morphine's analgesic response, shifting morphine's ED50 from 3.1 mg/kg alone to 0.86 mg/kg in conjunction with nefazodone (P less than 0.05). Two days after implantation of a morphine pellet (75 mg) no mice remained analgesic in the tailflick assay. Administration of nefazodone (50 mg/kg s.c.) restored analgesia to 60% of mice (P less than 0.03). In selective analgesic assays, nefazodone enhanced mu 1, mu 2 and delta analgesia, but not kappa 1 or kappa 3 analgesia. Nefazodone did not affect morphine's LD50 and, in assays of gastrointestinal transit, nefazodone increased morphine's potency only slightly. In conclusion, nefazodone alone is analgesic in certain animal models. In conjunction with morphine, nefazodone potentiated analgesia with no effect on lethality and little effect on gastrointestinal transit, resulting in an increase in morphine's therapeutic index. These results suggest that nefazodone and similar agents may have a significant role in the management of pain.


Effect of flosequinan upon isoenzymes of phosphodiesterase from guinea-pig cardiac and vascular smooth muscle. The effect of flosequinan and its sulphone metabolite BTS 53,554, on phosphodiesterase isoenzymes isolated from guinea-pig cardiac and vascular smooth muscle using DEAE-cellulose chromatography was investigated. Zaprinast and milrinone showed peak I and peak III selectivity, and IBMX non-selective activity respectively, against both cardiac and vascular smooth muscle isoenzymes, as expected for these reference inhibitors. Flosequinan and BTS 53,554 demonstrated non-selective inhibition with similar potency against both cardiac and vascular smooth muscle isoenzymes and, overall, were the least potent compounds tested. The high inhibitory concentrations observed (IC50 peak III 660 microM for cardiac tissue and 230 microM for vascular smooth muscle with flosequinan) relative to its clinically effective plasma concentration (10 microM) questions the relevance of phosphodiesterase inhibition to the efficacy of flosequinan in heart failure.


(R)-alpha-methylhistamine augments neural, cholinergic bronchospasm in guinea pigs by histamine H1 receptor activation. In the airways, activation of histamine H3-receptors with (R)-alpha-methylhistamine inhibits neurally induced cholinergic contractions in vitro and peptidergic responses in vivo. The role of histamine H3-receptors on the cholinergic bronchoconstriction induced by electrical stimulation of the dorsal medulla in guinea pigs was assessed in this study. There was no evidence for an H3-receptor mediated inhibition of cholinergic bronchospasm in vivo. However, there was potentiation of central cholinergic bronchoconstriction by (R)-alpha-methylhistamine or histamine by a mechanism involving H1-receptors. I.v. (R)-alpha-methylhistamine (0.3-3 mg/kg) or histamine (0.001-0.01 mg/kg) produced a transient bronchospasm and potentiated the bronchoconstriction due to medullary stimulation. These effects of (R)-alpha-methylhistamine and histamine were blocked by the histamine H1-antagonist, chlorpheniramine (30 micrograms/kg i.v.) but not by H2- or H3-receptor antagonists. (R)-alpha-Methyl-histamine did not potentiate the bronchoconstriction due to i.v. methacholine. Other bronchoconstrictor agents such as methacholine and serotonin did not potentiate the CNS-induced bronchospasm.


GABAA antagonists reveal binding sites for [35S]TBPS in cerebellar granular cell layer. Autoradiography revealed picrotoxin-sensitive [35S]TBPS ([35S]t-butylbicyclophosphorothionate) binding sites in the cerebellar granule cell layer only in the presence of GABAA receptor antagonists. However, under the same conditions, the binding was decreased in other brain regions, including the cerebellar molecular cell layer. Thus, the convulsant binding sites at GABAA receptors in the granule cell layer are highly sensitive to down-regulation by GABA, which may be due to a unique combination of receptor subunits.


Substrate specificity of diazepam-insensitive cerebellar [3H]Ro 15-4513 binding sites. [3H]Ro 15-4513, a negative modulator at the benzodiazepine receptor, binds with high affinity to all known benzodiazepine binding sites associated with the GABAA receptors. The present experiments surveyed a number of benzodiazepine receptor ligands with different chemical structures and different intrinsic pharmacological profiles for their ability to displace [3H]Ro 15-4513 binding in rat cerebellar membranes. The latter have been shown to possess two types of binding sites, one sensitive to positive modulators such as diazepam and the other insensitive to them (diazepam-insensitive). Whereas the full positive modulators from the benzodiazepine, beta-carboline, and quinoline classes did not displace binding at the diazepam-insensitive sites, the partial positive modulators' affinities for these sites varied much more, some being very potent and others having no affinity at all. All the neutral antagonists tested displaced binding at both types of binding sites, while some of the negative modulators were apparently not potent at the diazepam-insensitive sites. In an alcohol-sensitive rat line (ANT, alcohol non-tolerant), with an enhanced affinity for positive modulators at the diazepam-insensitive sites, these sites also exhibited enhanced affinity for some of the partial positive and negative modulators. The results suggest that the cerebellar diazepam-insensitive [3H]Ro 15-4513 binding sites have unique substrate specificity. It remains to be established which behaviours are affected by activation or inhibition of these receptors that are characterized by their insensitivity to benzodiazepine-positive modulators.


Evidence for GABAB-mediated mechanisms in experimental generalized absence seizures. Experimental absence seizures are characterized by the fact that they are exacerbated by both direct and indirect GABA agonists. To date most of the studies that have examined this phenomenon have utilized GABAA agonists. We assessed the effect of a GABAB agonist, baclofen and a specific GABAB antagonist in two pharmacological models of absence seizures in rodent after using either gamma-hydroxybutyrate or pentylenetetrazole to induce the bilaterally synchronous spike wave discharges that typify absence seizures in rodent. Baclofen markedly prolonged and the GABAB antagonist attenuated or blocked the experimental absence seizures in both models. These data suggest a role for GABAB-related mechanisms in the pathogenesis of generalized absence seizures and raise the possibility that GABAB antagonists may have therapeutic potential as antiabsence drugs.


Effect of Hoe 140, a new bradykinin receptor antagonist, on bradykinin- and platelet-activating factor-induced bronchoconstriction and airway microvascular leakage in guinea pig. We have investigated the effect of a new bradykinin receptor antagonist, Hoe 140 (D-Arg- Hyp3,Thi5,D-Tic7,Oic8]-bradykinin), on bradykinin- and platelet-activating factor (PAF)-induced bronchoconstriction and airway microvascular leakage in anesthetized guinea pigs. Extravasation of Evans blue dye and lung resistance were measured simultaneously. Both i.v. (15 nmol/kg) and inhaled bradykinin (1 mM, 45 breaths) caused a significant increase in lung resistance and leakage of dye at all airway levels. Hoe 140 (100 nmol/kg i.v.) almost completely inhibited these airway responses induced by bradykinin except for dye extravasation in trachea induced by inhaled bradykinin. Inhaled PAF (3 mM, 30 breaths) significantly increased lung resistance and leakage of due at all airway levels, but Hoe 140 had no effect on these responses. Bradykinin-induced bronchoconstriction and airway microvascular leakage are predominantly mediated by activation of B2 receptor, since Hoe 140 is a B2 receptor antagonist. Bradykinin receptor-mediated mechanisms do not play an important role on inhaled PAF-induced bronchoconstriction and microvascular leakage.


Effect of putative melatonin receptor antagonists on melatonin-induced pigment aggregation in isolated Xenopus laevis melanophores. The ability of putative melatonin receptor antagonists to antagonise melatonin-induced aggregation of pigment granules in cultured neural crest Xenopus laevis melanophores was examined. Neither ML 23 (N-(2,4-dinitrophenyl)-5-methoxytryptamine) nor 6-methoxy-2-benzoxazolinone showed agonist or antagonist activity. N-Acetyltryptamine and N-butanoyltryptamine were partial agonists; both compounds aggregated pigment granules in some cells, but also reversed melatonin-induced pigment agreggation in a fraction of the cells tested. In contrast, 2-benzyl N-acetyltryptamine (luzindole) did not show agonist activity (upto 10 microM) but did reverse the aggregating action of melatonin at 1 and 10 microM. Pretreatment of melanophores with luzindole shifted the melatonin concentration-response curve to the right.


Compound 48/80 blocks transmission and increases the excitability of ganglion neurons. Compound 48/80 (5.0-50 micrograms/ml) significantly and reversibly decreased (1) the amplitude, but not the shape of the compound action potential, (2) the amplitude and duration of the acetylcholine potential and (3) the residual fast excitatory postsynaptic potential recorded from neurons of the 9th and 10th paravertebral ganglia of the bullfrog Rana catesbeiana. The excitability of B-type ganglion neurons in the presence of nicotinic and muscarinic receptor antagonists was increased by compound 48/80 without altering the input resistance or membrane potential. In addition, compound 48/80 (10-50 micrograms/ml) significantly decreased the duration of the spike afterhyperpolarization (AHP). The amplitude but not the decay rate of the current underlying the slow component of the spike AHP was decreased by compound 48/80. Compound 48/80 did not, however, alter either the amplitude or the duration of calcium-dependent spikes. Intracellular recordings from dissociated sympathetic neurons also demonstrated a compound 48/80-induced increase in neuronal excitability. These results suggest that compound 48/80 interacts with the nicotinic receptor/channel complex to decrease ganglionic transmission, and also has a direct action to increase neuronal excitability by blocking potassium channels mediating the duration of the spike AHP.


Polyamines enhance calcium mobilization in fMet-Leu-Phe-stimulated phagocytes. Spermidine and putrescine (50 microM-1 mM), found in exudates from infection sites, significantly enhance fMet-Leu-Phe-induced Ca2+ mobilization in differentiated HL-60 cells and polymorphonuclear leukocytes (PMNs) by delaying the return to basal cytosolic Ca2+ levels. This enhancement by polyamines is associated with inhibition of Ca2+ efflux across the plasma membrane. In parallel with their effects on Ca2+ signaling, polyamines also significantly prolong the kinetics of fMet-Leu-Phe-induced protein kinase C translocation. Thus, polyamines may play a novel role in modulating regulatory events in phagocytes.


Enzymatic activity of adenylate cyclase toxin from Bordetella pertussis is not required for hemolysis. Adenylate cyclase (AC) toxin from Bordetella pertussis enters cells to cause supraphysiologic increases in cAMP. AC toxin is also hemolytic. Substitution of Lys-58 with a methionine residue by site-directed mutagenesis of the structural gene for AC toxin, cyaA, and introduction of this mutation onto the B. pertussis chromosome results in an organism that synthesizes an enzyme-deficient AC toxin molecule. This mutant toxin molecule exhibits 1000-fold reduction in enzymatic activity relative to wild-type and has no toxin activity in J774 cells. The enzyme-deficient toxin molecule is not, however, impaired in its ability to lyse sheep red blood cells. In order to ascertain the importance of these two separate activities of AC toxin in vivo the enzyme-deficient organisms were used to infect infant mice. The hemolytic, enzyme-deficient mutant organisms are reduced in virulence relative to wild-type organisms after intranasal challenge indicating that, although the enzymatic activity of AC toxin does not contribute to hemolysis, it is this property of the toxin which is important for virulence of B. pertussis.


Neopterin as an endogenous antioxidant.

Neopterin as an endogenous antioxidant. The in vitro potency of neopterin (NP) as an antioxidant and its in vivo activity to suppress alloxan-induced diabetes were investigated. The reduced form of neopterin, 5,6,7,8-tetrahydroneopterin (NPH-4), showed an extremely high superoxide anion radical scavenging activity in two assay systems, i.e. xanthine/xanthine oxidase- and macrophage/phorbol myristate acetate (PMA)-reaction systems. NPH-4 also inhibited the oxidation of linoleic acid about as effectively as uric acid. Furthermore, NPH-4 and NP effectively suppressed alloxan-induced mouse diabetes. These results suggest that pteridines play an important role as endogenous antioxidants.


Heterogeneous expression of four MAP kinase isoforms in human tissues. Mitogen-activated protein kinases (MAP kinases) are a group of closely related enzymes implicated in signal transduction pathways. We report the molecular cloning of four human proteins (p40mapk, p41mapk, p44mapk and p63mapk) with high homology to members of the MAP kinase family. Sequence analysis demonstrated that p44mapk and p63mapk were the products of distinct genes. However, the p40mapk and p41mapk were found to be related, and are likely to result from alternative processing of transcripts from a single gene. The heterogeneous expression of these human MAP kinase isoforms in different tissues may reflect the diversity of signal transduction pathways in differentiated cells.


Ca(2+)-dependent translocation of cytosolic proteins to isolated granule subpopulations and plasma membrane from human neutrophils. In order to identify cytosolic proteins involved in control of granule exocytosis in human neutrophils, subcellular fractions enriched in each of the 3 major granule subsets were incubated with cytosol from neutrophils in the presence or absence of Ca2+. After washing, proteins were eluted from the organelles by EGTA. Annexins I, II, IV and VI were found to bind to all organelles studied. In addition, a 28-kDa protein was found to bind exclusively to plasma membranes and secretory vesicles, the most readily exocytosed organelle of neutrophils. Ca(2+)-dependent association of cytosolic proteins to different granule subsets may control differential exocytosis of granules.


Activation of type-1 protein phosphatase by cdc2 kinase. Purified cdc2 or cdc2 obtained from HeLa cells in association with p13suc1 activate inactive type-1 protein phosphatase (PP1) (catalytic subunit.inhibitor-2 complex, purified from skeletal muscle). Likewise in the case of PP1 activation by FA/GSK3, activation by cdc2 is accompanied by phosphorylation of inhibitor-2 (I2) and free I2 can be phosphorylated as well. Correlation between PP1 activation and I2 phosphorylation is suggested by the fact that both activation and phosphorylation (a) increase in parallel during incubation with cdc2, (b) decrease in parallel upon subsequent cdc2 inhibition by EDTA, and (c) are inhibited by the cdc2 inhibitor 5,6-dichlorobenzimidazole riboside. cdc2 also phosphorylates the catalytic subunit of PP1, whether in the complex with I2 or as free molecule. The activation of PP1 by cdc2 and by FA/GSK3 is compared.


A rapid calcium influx during exocytosis in Paramecium cells is followed by a rise in cyclic GMP within 1 s. The synchrony of trichocyst exocytosis in Paramecium allows temporal correlation of associated events. Using quenched flow we observed a Ca2+ influx concurrent with exocytosis within 80 ms after stimulation with the secretagogue aminoethyldextran. Cyclic AMP did not change in depency of stimulation. Cyclic GMP transiently increased after 500 ms, culminating at 2 s, and thus considerably lags behind exocytosis induction and influx of Ca2+. Both Ca2+ influx and rise in cGMP are known to be induceable also by Ba2+ or veratridine, allegedly via the opening of ciliary Ca2+ channels. However, only veratridine stimulated exocytosis. We conclude that both aminoethyldextran and veratridine induce an exocytosis-associated Ca2+ influx, which is responsible for the rise in cGMP, through an as yet unknown pathway.


Phosphorylation of the phosphatase modulator subunit (inhibitor-2) by casein kinase-1. Identification of the phosphorylation sites. The isolated modulator subunit of the inactive protein phosphatase-1 is phosphorylated in vitro by casein kinase-1 at two different sites: Ser-86 and Ser-174. The Ser-86 site is a common target for casein kinase-1 and casein kinase-2, but is preferentially phosphorylated by the former enzyme. The Ser-174 site seems to be specific for casein kinase-1, and is phosphorylated at a slower rate. These results give a new insight into the in vitro phosphorylation pattern of the modulator subunit of the phosphatase and provides additional data on the specificity of casein kinase-1.


Minimal functional size of porcine lung and testicular angiotensin-converting enzymes deduced from radiation inactivation analysis. Interaction of two highly homologous domains in somatic isoenzyme. Domain structures of porcine lung and testicular angiotensin-converting enzymes (ACE) were studied by radiation inactivation to test the hypothesis that lung ACE has two catalytic sites localized to discrete, structurally independent domains (the N- and C-domains) of approximately equal size. The minimum functional sizes of lung and testicular ACE, calculated from the inactivation curves obtained, were 140 and 74 kDa, respectively. Since testicular ACE has been demonstrated to contain only the C domain, this result indicates that the two domains in lung ACE are not independent but are, in fact, structurally tightly linked.


The effect of the UL42 protein on the DNA polymerase activity of the catalytic subunit of the DNA polymerase encoded by herpes simplex virus type 1. The effect that the UL42 protein of herpes simplex virus type 1 has on the DNA polymerase activity of the DNA polymerase catalytic subunit (Pol) of the same virus has been investigated. The observed effects are critically dependent on the salt used and its concentration, such that the UL42 protein may inhibit, have little or no effect on, or activate the Pol activity, depending on the condition used. The observed effects are due to the values for Km(app) for activated DNA and Vmaxapp for Pol and the Pol-UL42 protein complex differently varying with salt concentration.


Cyclic AMP and Ca2+ as regulators of zygote formation in the cellular slime mold Dictyostelium mucoroides. To analyze the mechanism of the sexual process (macrocyst formation) in the cellular slime mold Dictyostelium mucoroides-7 (Dm7), the effects of 3',5'-cyclic adenosine monophosphate (cAMP), conditioned medium (CM) factors, and various ions including Ca2+ on zygote formation were examined. The application of cAMP was found to inhibit the sexual cell fusion. In addition, the activity of fusion inhibitor(s) contained in CM was heat stable and lost by phosphodiesterase (PDE)-treatment, thus indicating that cAMP is the inhibitor, being in contrast to ethylene as a fusion activator. Pulse experiments using two cAMP analogues, 2'-deoxy-cAMP and 8-bromo-cAMP suggested that the signal transduction system through the cell surface cAMP receptor is of particular importance for regulation of the sexual fusion process. Among several ions having effects on zygote formation, Ca2+ seemed to be necessary both for the acquisition of fusion competence and for cell fusion itself. In the presence of Ca2+, K+ and Na+ had the opposite effects on zygote formation; K+ was stimulative, while Na+ inhibitory. The significance of these findings is discussed in relation to the regulatory mechanism of zygote formation.


Morphogenesis and differentiation of Dictyostelium cells interacting with immobilized glucosides: dependence on DIF production. Previous work has shown that multicellular morphogenesis of submerged Dictyostelium cells is inhibited when they bind to glucosides covalently linked to polyacrylamide gels. The amoebae aggregate normally, but then the aggregates repeatedly disperse and reaggregate, whereas control cells go on to form tight aggregates. We have investigated the role of the stalk cell differentiation inducing factors (DIFs) in this process. In the presence of cyclic AMP, amoebae submerged at high cell density accumulate DIF and differentiate into stalk cells. We find that stalk cell differentiation is inhibited by interaction of the cells with glucoside gels in these conditions, but can be restored by the addition of exogenous DIF-1. Since the responsiveness of cells to DIF-1 is not altered, it appears likely that the effect of the glucoside gel is to block DIF-1 production. Further, the addition of DIF-1 or DIF-2 stimulates the formation of tight aggregates by cells developing on glucoside gels in the absence of cyclic AMP, thus preventing the rounds of aggregation and disaggregation otherwise seen. This suggests a role for DIF in morphogenesis as well as in controlling cell differentiation. We propose a model in which immobilized glucosides activate a specific receptor ("food sensor") which drives the amoebae toward the vegetative state and inhibits DIF accumulation. DIF, on the other hand, induces tight aggregate formation and so locks the amoebae into the developmental program.


Medication counseling for the elderly: effects on knowledge and compliance after hospital discharge. The findings from the instruments used in this study and the additional qualitative data of the personal interviews provide information helpful to the nurse's understanding of medication knowledge and compliance patterns in older persons. Investigations that study older persons over time are needed to identify how behaviors change. Such studies would also be helpful in identifying self-care behaviors learned over extended periods of time. It may also be helpful to investigate knowledge and compliance behaviors in the home setting, where the environment may be more conducive to learning. With the patient assuming more and more responsibility for self-care, it is imperative that professionals explore all opportunities to assist patients in acquiring the skills essential to promoting their own well-being.


The elderly patient receiving radiation therapy: treatment sequelae and nursing care. Preparing the elderly patient for radiation therapy requires knowledge of the major cancers, the role of radiation treatment, sequelae of treatment, and the age-associated factors that exacerbate treatment reactions. Elderly patients often require long treatment courses. Encouragement to express feelings and report symptoms may be necessary in this patient population. Instructions should be written and repeated, especially for patients with memory loss. The nurse caring for the elderly patient receiving radiation must help the patient to anticipate side effects and manage sequelae of treatment. The patient who knows what to expect is better able to tolerate therapy and to maintain an optimal quality of life during treatment.


Reversible binding of the cooked food mutagen MeIQx to lignin-enriched preparations from wheat bran. The binding of the mutagen 3,8-dimethyl-3H-imidazo[4,5-f]quinoxaline-2-amine (MeIQx) to various fibre preparations from wheat bran was studied. The physical structures of wheat bran and lignin-enriched preparations were determined by scanning electron microscopy. With increasing mutagen concentration from 0.5-16 micrograms/ml, the fraction of MeIQx bound to cellulase-treated lignin was nearly constant, for a certain lignin density. The binding between cellulase-treated lignin and MeIQx was reversible. Incubation temperature influenced the rate at which the equilibrium between lignin and MeIQx was established, but had less effect on the equilibrium itself. With increasing fibre densities and a constant mutagen concentration, complete binding was apparently reached at a high fibre density. This was illustrated by plotting the data according to Scatchard. Increased binding of the MeIQx was obtained with increased enrichment of lignin in different fibre preparations.


Effect of murine retroviral infection on hair and serum levels of cocaine and morphine. LP-BM5 retrovirally infected female C57BL/6J mice were administered cocaine, morphine or both by daily intraperitoneal injection for 9 weeks. Drug concentrations were measured by radioimmunoassay in serum and in hair extracts. Hair samples obtained from all drug-treated mice were positive for the drug injected, while none of the saline-treated mice had detectable drug levels in hair or serum. The average morphine concentration obtained from non-infected mice was 11 ng/mg hair whereas the amount found in the LP-BM5-infected mice was significantly higher (20 ng/mg hair). Mice injected with both morphine and cocaine were given 50% of the regular dose of each drug and drug levels in the hair of these animals were approximately half that of mice injected with the full dose of the single drug. Non-infected mice treated with both drugs had a mean value of 7 ng morphine/mg hair and 374 ng cocaine/mg hair while retrovirus-infected mice had significantly higher concentrations, 10 ng morphine/mg hair and 1160 ng cocaine/mg hair (P less than 0.001). Serum concentrations of cocaine and morphine were significantly higher (P less than 0.01) in the retrovirus-infected animals from 40 min to 1.5 h. The increased concentrations of cocaine and morphine in serum during retrovirus infection are accompanied by a significant increase in the amount of drug incorporated into the hair matrix. This change indicates that retroviral infection may influence the disposition of these drugs in the systemic circulation.


Morphological differences between hepatocellular carcinoma and hepatocellular carcinomalike lesions. In an attempt to provide a quantitative basis for differentiation between well-differentiated hepatocellular carcinoma and hepatocellular carcinomalike lesions (focal nodular hyperplasia, regenerative nodular hyperplasia and hepatocellular adenoma), histopathological and morphometrical analyses were performed on 208 cases of various liver diseases with the aid of an image analyzer. As practical indicators for hepatocellular carcinoma, the following six morphometrical features were established: (a) nuclear shape factor of less than 0.93, (b) coefficient of variance of nuclei of more than 5%, (c) average width of trabecular cords greater than three cells, (d) nucleocytoplasmic ratio increased to more than 0.3, (e) cellular density of more than 40 liver cells and (f) individual nuclear dimension larger than 50 microns2. The manifest categories increased with dedifferentiation of cells in hepatocellular carcinoma; the number and degree of cellular and structural atypia became more prominent. Data were analyzed statistically by two multivariate analyses. Logistic analysis was able to correctly separate hepatocellular carcinoma from conditions that were not hepatocellular carcinoma, including hepatocellular carcinomalike lesions. The incidence of 13 descriptive histopathological findings such as fibrous capsule, portal triads and clear-cell clusters were also compared in hepatocellular carcinomalike lesions and hepatocellular carcinomas graded according to Edmondson's classification. Presence of a fibrous capsule, portal triads, mosaiclike patterns and tumor vessels showed statistical differences between hepatocellular carcinomalike lesions and well-differentiated hepatocellular carcinoma by the chi 2 test (p less than 0.005).