Somatostatin in acute bleeding oesophageal varices. Pharmacology and rationale for use. Somatostatin was originally isolated from the hypothalamus, but has subsequently been found throughout the whole gastrointestinal tract. It exerts an inhibitory effect upon numerous functions of the body, and, therefore, increasing attention has been focused on its potential as a therapeutic agent with cytoprotective properties and a potent inhibitory action on a wide variety of functions in endocrine diseases, cancer and gastrointestinal haemorrhage, including bleeding from oesophageal varices. As somatostatin has a very short plasma half-life and requires administration by continuous infusion to maintain therapeutic levels, stable long-acting analogues have been developed. The analogue octreotide has been shown to have a plasma half-life of 113 minutes and to produce a profound selective inhibition of growth hormone. Hepatic excretion of octreotide has been estimated to be between 30 and 40% in healthy volunteers; no data are available in cirrhotic patients. A review of published data assessing systemic and hepatic haemodynamics in animals and humans over varying dosage regimens of somatostatin and octreotide reveals that cardiac output, arterial pressure and peripheral resistance are modified more in animals than in humans. Hepatic haemodynamics are significantly altered in animals as well as in humans in most of the studies. Circumstantial evidence is provided indicating that the mechanisms of action of somatostatin and octreotide in the therapy of bleeding oesophageal varices are mainly mediated by a splanchnic vasoconstrictive effect. Furthermore, gastric acid suppression and potential enhancement of platelet aggregation may contribute to the beneficial outcome after treatment of oesophageal varices with somatostatin.


Somatostatin in acute bleeding oesophageal varices. Clinical evidence. Following the demonstration that somatostatin lowered portal pressure in cirrhotic patients with portal hypertension, 2 uncontrolled reports suggested that the hormone might be useful in the control of acute variceal haemorrhage. Subsequently, a number of randomised controlled trials have indicated that somatostatin may have an efficacy as good as or better than either vasopressin or combined vasopressin and nitroglycerin therapy and is associated with fewer side effects. Somatostatin has an efficacy comparable to balloon tamponade, histamine-2-receptor antagonists and injection sclerotherapy. One double-blind randomised controlled trial demonstrated a significant benefit of somatostatin over placebo in the control of variceal bleeding whereas a second did not show any significant difference between treatments. In all the controlled trials, the average control rate achieved with somatostatin administration was 69% and it was not associated with any major side effects. Somatostatin administration has also been shown in uncontrolled series to be very effective in controlling postinjection sclerotherapy bleeding from the varices per se, and from oesophageal ulcers and oesophagitis. Few data are available on the long acting analogue of somatostatin, octreotide, but preliminary data suggest that it may be as effective and safe as the native hormone in controlling the acute variceal bleeding and postinjection sclerotherapy haemorrhage. It is concluded that there may be a case for instituting somatostatin therapy as soon as the patient enters hospital to facilitate sclerotherapy, and for continuing treatment for 5 days after sclerotherapy when the risk of recurrent bleeding is highest.


Effects of benzo[a]pyrene on steady-state levels of biogenic amines and metabolizing enzymes in mouse brain regions. Benzo[a]pyrene (BaP) is a product of incomplete fossil fuel combustion, a well-known pollutant, and a carcinogenic agent. In the present study male CD-1 mice received ip injections of 0, 5, 25, and 100 mg/kg body weight BaP twice a week for 3 weeks. Endogenous levels of brain biogenic amines and their selected metabolites, norepinephrine (NE), dopamine (DA), 5-hydroxytryptamine (5-HT), vanillylmandelic acid, dihydroxyphenylacetic acid (DOPAC), homovanillic acid, and 5-hydroxyindoleacetic acid (5-HIAA) were measured using high performance liquid chromatography and electrochemical detection. The brain regions studied were cortex, striatum, hypothalamus, midbrain, medulla oblongata, and cerebellum. BaP treatment increased the steady-state levels of NE, DA, and 5-HT in the hypothalamus and striatum. Increased levels of DA and 5-HT and their major metabolites DOPAC and 5-HIAA were noticed in the same region, an indication of increased metabolism of these amines. The increase in the 5-HT level in the cortex was not dose-related. Levels of NE and DA were significantly higher in the medulla oblongata. There was a concurrent increase in activities of tyrosine hydroxylase and tryptophan hydroxylase in several brain regions. The effect of BaP on Dopa-decarboxylase was not consistent. Monoamine oxidase was occasionally inhibited. Results indicate that exposure to BaP altered the steady-state levels of biogenic amines in various brain regions and these changes were consistent with the activities of metabolizing enzymes.


Long term management of oesophageal varices. Effective control of variceal rebleeding (secondary prophylaxis) or prevention of the initial bleeding (primary prophylaxis) are the main objectives of the treatment of portal hypertension. Endoscopic sclerotherapy is the treatment of choice for secondary prophylaxis, since it significantly decreases rebleeding and, to some extent, mortality. A combination of propranolol and sclerotherapy may be of benefit by decreasing postsclerotherapy rebleeding. Endoscopic variceal band ligation and transjugular intrahepatic shunt are emerging as useful alternative techniques. Devascularisation and preferably selective shunts should be reserved for use as salvage of sclerotherapy failures. Liver transplantation, if feasible, could become the ultimate therapy by controlling variceal bleeding and improving hepatic function. Pharmacotherapy, while not very successful for secondary prophylaxis, has shown promise for primary prophylaxis of variceal bleeding. Nonselective beta-blockers significantly decrease the rebleeding rates but are associated with only marginal survival benefits. beta-Blockers alone cannot decrease the hepatic venous pressure gradient adequately (to less than 12mm Hg). Combination with nitrates and other drugs may prove beneficial and requires clinical evaluation. Endoscopic sclerotherapy and surgery have little role in primary prevention of variceal bleeding in patients with cirrhosis but need evaluation in noncirrhotic patients.


Trace metal accumulation by the shrew Sorex araneus. I. Total body burden, growth, and mortality. A laboratory feeding trial is described in which a known trace metal accumulator, the shrew Sorex araneus, was presented with an artificial diet containing high levels of copper and cadmium. Methodological difficulties relating to feeding trials on first generation offspring of insectivorous small mammals caught in the wild are described. There was no relationship between copper or cadmium intake and mortality during the trials. Sex differences in growth were apparently not influenced by metal intake. At the end of the trials there was a significant negative correlation between body weight and total cadmium burden. Whole body concentrations and burdens of cadmium increased with estimated intake. There was a significant negative correlation between percentage of cadmium retained as body burden and estimated cadmium intake. There was no discernible relationship between copper burden and intake.


Trace metal accumulation by the shrew Sorex araneus. II. Tissue distribution in kidney and liver. The distributions of trace metals in tissues of shrews in a series of feeding trials are described. The relationship between body cadmium burden and organ weight was directly related to differences in body weight. Organ weight expressed as a percentage of body weight was constant in the kidney, but was correlated with cadmium burden for the liver. Liver and kidney accounted for up to 80% of total body cadmium. The proportion of the body cadmium burden found in the kidney ranged from 8 to 17%. Copper and zinc concentrations in the kidney increased in response to cadmium accumulation. Hepatic zinc concentrations increased with cadmium accumulation. Exposure to a copper-contaminated diet resulted in increased liver copper levels. High liver concentrations were correlated with high kidney concentrations. At the lowest levels of exposure to metals liver and kidney concentrations were similar, but as exposure increased so did liver concentrations and much more so than in the kidney.


Investigations of the acute toxic, cytogenetic, and embryotoxic activity of buminafos. The organophosphorus herbicide buminafos (O,O-dibutyl-(1-butylaminocyclohexyl)-phosphonate) was tested for its acute toxic, cytogenetic, and embryotoxic activity on different strains of mice. The oral LD50 value for male NMRI mice was determined to be 3500 mg/kg. Single oral doses of 175, 1000, and 2000 mg/kg did not cause any significant enhancement in the percentage of chromosome aberrations in bone marrow cells of male NMRI mice. After oral administration of 500 and 1000 mg/kg buminafos to pregnant Halle:DBA and Halle:AB mice at Days 6-15 of gestation no embryotoxic effects were observed. The cytogenetic inactivity of buminafos in the bone marrow chromosome assay corresponds to negative findings in other mutagenicity tests.


Thyroid physiology impairment by malathion in the freshwater catfish Clarias batrachus. Effects of malathion on serum and glandular thyroxine (T4), triiodothyronine (T3), the T3/T4 ratio, peroxidase activity, and extrathyroidal conversion of T4 to T3 have been studied in the freshwater catfish Clarias batrachus during the prespawning and spawning phases of its annual reproductive cycle. Malathion inhibited T4 secretion in the kidney, but accelerated T4 synthesis in the pharyngeal thyroid. This pesticide also inhibited the extrathyroidal conversion of T4 to T3 in serum during both the studied phases.


Effects of polyoxyethylene (20) sorbitan monooleate on the acute toxicity of linear alkylbenzenesulfonate (C12LAS) to fish. Effects of polyoxyethylene (20) sorbitan monooleate (SMOE20) on the acute toxicity of linear alkylbenzenesulfonate (C12LAS) to fish were investigated in red killifish (Orizias latipes) and carp (Cyprinus carpio). By adding polyoxyethylene sorbitan ester to C12LAS solution, the acute toxicity of C12LAS decreased. As a decreasing toxic effect of these nonionic surfactants, the depression of gill damage was histopathologically observed. No significant hematological and blood biochemical differences were observed between two tests using a single C12LAS solution and a mixed solution of C12LAS/SMOE20. The addition of SMOE20 to C12LAS decreased the incorporation of C12LAS into blood and each organ except spleen and gall bladder as well as the adsorption of C12LAS to the gill. The farther inward the phenyl position in the alkyl side chain, the lower the adsorption of C12LAS to gill. Since the acute toxicity of LAS phenyl isomers to fish was known to decrease according to the order of adsorption, these results suggest that the decreasing toxic effect of SMOE20 on the acute toxicity of C12LAS is due to depression of more toxic C12LAS adsorption on the gill.


Mercury concentration correlates with the nitrogen stable isotope ratio in the animal food of Papuans. The relationships among element concentrations (Na, Mg, Al, P, K, Ca, Cr, Fe, Mn, Cu, Zn, Sr, total Hg, organic Hg, inorganic Hg, Pb) and stable carbon and nitrogen isotope ratios (13C/12C and 15N/14N) in animals consumed by the people called Gidra, who inhabit the lowland of Papua New Guinea, were examined. Animals analyzed included mammal, bird, fish, shellfish, reptile, crustacean, and insect. Highly significantly positive correlations were observed between total Hg concentrations and 15N/14N (r = 0.796), between organic Hg concentrations and 15N/14N (r = 0.781), and between inorganic Hg concentrations and 15N/14N (r = 0.739). This was interpreted to indicate that Hg was an element which accumulates in animals along the food chain. Based on the regression function of Hg on delta 15N, the bioconcentration factor for total, organic, and inorganic Hg was estimated to be 5.


Effects of coal gasification slag as a substrate for the plant Cyperus esculentus and the worm Eisenia fetida. A further development of the coal gasification process will result in an increase of the amount of coal gasification slag (CGS). As yet little is known about the effects of storage in uncovered dumps. If there are any environmental effects, they are most likely caused by accumulation of metals from the CGS or by unacceptable physical properties of the CGS. Growth inhibition, mortality, and metal accumulation were analyzed for the plant Cyperus esculentus and the worm Eisenia fetida on CGS substrate. Pulverized fuel ash (PFA) was used as a reference. Both in the substrate and in tissues the concentrations of the cations Cu, Ni, Pb, and Zn and the anions As, B, Cr, Mo, Sb, and Se were determined. The availability of anions for C. esculentus and for E. fetida is greater in PFA than in CGS. The extent and rate of uptake of anionic metals by the plants is on the whole higher in the wetland situation. The availability of metals, expressed as the concentration factor (CF), in most cases appears to be smaller than 1 for nearly all elements. In E. fetida a CF greater than 1 was found only for the element As, in PFA substrate and 50% CGS. In C. esculentus a CF greater than 1 was found for B and Mo in the PFA substrate as well.


Frozen algae as food for Daphnia magna Straus in toxicity testing. As EC validity criteria for the production of Daphnia magna neonates in standard toxicity tests set fecundity minima which must be attained, it is important that culture conditions provide an adequate ration of high quality food. Maintaining a steady supply of fresh algae can be problematical so the possibility of using frozen food was investigated. The performance of three generations of D. magna fed on diets of fresh or frozen Chlorella was compared. There were no significant effects on fecundity due to food type or generation. Cumulative fecundity (over 21 days) on fresh or frozen food exceeded 120 neonates per female and satisfied that EC validity criterion (60 neonates per female). In addition criteria relating to survival and variability were met. We conclude that frozen Chlorella are an acceptable substitute for fresh algae.


Toxic effects of bleached and unbleached paper mill effluents in primary cultures of rainbow trout hepatocytes. Toxic effects of unbleached (sulfate or sulfite) and bleached (sulfate) paper mill effluents were studied in a primary culture of rainbow trout liver cells. The effluents and control water from a clean area were extracted with diethyl ether and added to the cultures dissolved in dimethyl sulfoxide. Plasma membrane integrity was studied by measuring lactate dehydrogenase (LDH) leakage. The cellular content of glutathione (GSH) was used as an indicator of oxidative stress and the formation of reactive intermediates. Dose-response studies indicated that unbleached effluents contained more potent toxic substances than bleached effluents. Both unbleached and bleached effluents contained organic diethyl ether-extractable substances which increased cytochrome P450-dependent 7-ethoxyresorufin-O-deethylase (EROD) activities. The inducing effects were seen at concentrations substantially lower than those decreasing GSH content and increasing LDH leakage. Possible EROD inducing substances in bleached effluents are chlorinated organic compounds. Inducing compounds in unbleached effluents are yet to be identified. Furthermore, at higher concentrations the effluents contained substances that inhibited the cytochrome P450 system. The results show that the trout primary hepatocyte cultures afford a convenient in vitro method for screening cytochrome P450 inducing components extracted from industrial effluents to investigate mechanisms by which wastewaters cause injury in cells.


Preexposure temperature acclimation and diet as modifying factors for the tolerance of golden ide (Leuciscus idus melanotus) to short-term exposure to 4-chloroaniline. The influence of different temperature and nutrition regimes on the acute toxicity of 4-chloroaniline to golden ide (Leuciscus idus melanotus) was investigated. Acute toxicity was determined over 48 hr at 20 degrees C without feeding after a 2-day acclimation period. In an attempt to reveal underlying mechanisms accounting for diet- and temperature-related differences in the toxicant resistance of golden ide, biochemical and quantitative morphological parameters of the liver, a central organ in the xenobiotic metabolism of fish, were recorded throughout the 12-week acclimation period. In cold-acclimated fish, acute toxicity of 4-chloroaniline was 40% higher than in fish acclimated to 20 degrees C. If compared to 20 degrees C, preacclimation to 14 degrees C was characterized by a lower specific growth rate, an increase of hepatic glycogen, and a decrease of body and liver lipid deposits. The organelle composition of hepatocytes was not significantly altered by temperature acclimation. For the nutrition experiment, commercially available diets A and B of similar crude composition were used. Acute toxicity of 4-chloroaniline was 60% lower with diet B than with diet A. If compared to diet B, diet A induced a higher specific growth rate and increased hepatocellular volume of endoplasmic reticulum and Golgi fields, whereas glycogen and lipid of the liver as well as body lipid contents were reduced. The toxicity of 4-chloroaniline was correlated with the development of the endoplasmic reticulum, the major site of biotransformation enzymes. A consistent correlation with lipid contents could not be established. Results illustrate not only that assay conditions during the actual test may profoundly interact with results of toxicity studies, but also that maintenance conditions before the test can have significant consequences on results. In order to improve reproducibility of the results of acute toxicity tests, more consideration should be given to the standardization of pretest maintenance conditions of fish.


Organomercury determination in biological reference materials: application to a study on mercury speciation in marine mammals off the Faröe Islands. The potential use of graphite furnace atomic absorption spectrometry (GF-AAS) for the organic mercury determination in marine biological tissues was evaluated. Following its isolation by acid extraction in toluene, organic mercury was recovered in aqueous thiosulfate and measured by GF-AAS. The detection limit was 0.01 microgram Hg/g (as methyl mercury). Analyses were conducted on three reference standard materials certified for their methyl mercury content, DOLT-1, DORM-1, and TORT-1, provided by the National Research Council of Canada. The method resulted in very good recovery and reproducibility, indicating that GF-AAS can provide results comparable to those obtained by using more expensive and time consuming analytical techniques. The method was applied to the analysis of liver tissues of pilot whale specimens (Globicephala melas) from the drive fishery of the Faröe Islands (northeast Atlantic). The results provided useful information on the proportion of different mercury forms in the liver of these marine mammals.


Automated interictal EEG spike detection using artificial neural networks. Feed-forward, error-back-propagation artificial neural networks were applied to recognition of epileptiform patterns in the EEG. The inherent network properties of generalization and variability tolerance were effective in identifying wave forms that differed from the training patterns but still maintained 'epileptiform' spatio-temporal characteristics. The certainty of recognition was measured as a continuous function with a range of 0-1. Two levels of certainty (0.825 and 0.900) were used to indicate recognition of spikes and sharp waves (SSW). An average 94.2% (+/- 7.3) of the SSW were recognized; 20.9% (+/- 22.9) of all recognized SSW were false-positive recognitions. The time required for pattern recognition was well within the time required for digitizing the analogue data. This study provides evidence that neural network technology is, in principle, an effective pattern recognition strategy for identification of epileptiform transients in the EEG. The analysis is sufficiently rapid to be of potential value as a strategy for data reduction of long recordings stored on bulk media.


Development of epileptic activity induced by iron injection into rat cerebral cortex: electrographic and behavioral characteristics. Unilateral injection of ferrous chloride solution into the rat sensorimotor cortex produced epileptic discharges in the electrocorticograms (ECoGs). The discharges were isolated spikes and spike and wave complexes, and the epileptic activity lasted for more than 12 months after the injection. Isolated spike activity often appeared on the left or right side of the cortex, whereas spike and wave complex activity appeared bilaterally. In rats showing dominant isolated spike activity in the secondary epileptic cortex, there was a deviation in somatosensory evoked potentials (SEPs). Rats showing isolated spikes and spike and wave complexes exhibited vibrissa tremors and head nodding. Rats showing only isolated spikes exhibited no abnormal behavior, but their convulsion thresholds to pentylenetetrazol were lowered. The results including ECoGs, SEPs, behavior and convulsion threshold were characterized with reference to the development of iron-induced epilepsy. The profiles of ECoG discharge activity and SEP configuration suggest that the process of iron-induced epilepsy consists of 3 stages.


Ontogenesis of nocturnal organization of sleep spindles: a longitudinal study during the first 6 months of life. Ontogenesis of sleep spindles was studied on overnight longitudinal recordings in 12 full-term infants at 1.5-3-4.5 and 6 months of life. Six parameters (density, duration, frequency, amplitude, asymmetry and asynchrony) were analyzed during both slow wave sleep (SII and delta) and during 5 periods of the night. Results show a significant increase of most parameters between 1.5 and 3 months of age. All spindle patterns developed quite rapidly during the first 3 months of infancy, possibly reflecting developmental changes in thalamo-cortical structures and maturation of the physiological system that produces spindles. The density of 12-14 Hz spindle frequency was higher in stage II when compared to stage delta, as in adults. Our data confirm previous reports on spindle ontogenesis and give a more complete aspect of this ontogenesis in relation to sleep development. Three months of age appeared to be a turning point in maturational processes and might reflect changes in central nervous system activity and behavior which take place during that period. Sleep spindle evolution seems to be an accurate reflection of the slow wave sleep (SWS) development, and our results are discussed in terms of the developmental aspect of SWS production and characterization of sleep stages in young infants. Concordance between quantitative aspects and nocturnal organization leads us to consider that the individualization of slow wave sleep (SWS) in infants occurs from 4.5 months of life.


Changes in cortical negative DC shifts due to different motor task conditions. The experiments were performed to study the relationship between motor performance and DC potential curves recorded by scalp electrodes. Accordingly, we studied the influence of different movements (e.g., unilateral versus bilateral, simple versus complex, active versus passive, phasic versus tonic muscle activity) on negative DC potentials. Our results confirm that spatial distributions of DC potential maxima can be used as an indicator of the activation of distinct cortical areas. Furthermore, evidence is presented that some motor tasks have a greater influence on the magnitude of surface electronegativity than others. (1) Phasic muscle activity revealed a significantly larger potential size than tonic. (2) Performance of a complex finger movement task elicited an increased surface electronegativity compared with performance of a simple task. (3) No significant differences in potential size were found between left (untrained) and right (skilled) hand use during the performance of the same complex motor task. (4) This was also true for the performance of an active and a passive finger movement task, indicating that, at least in simple motor tasks, somatosensory afferents significantly contribute to the recorded potential curve.


Event-related potentials to repetition and change of auditory stimuli. The major intent of this study was to compare the role of stimulus repetition and change in the elicitation of the MMN, an ERP component specific to stimulus change, and N2b, usually partially overlapping the MMN when stimuli are attended. Event-related potentials were recorded in one set of conditions where subjects ignored the stimuli and read a book, and in another set of conditions where subjects counted stimuli designated as targets. Stimuli were delivered in 4 ways, the common feature between all these conditions being the occurrence of infrequent events at a probability of 0.20: (1) an oddball paradigm with 1 deviant, (2) an oddball paradigm with 2 deviants, each with a probability of 0.10, (3) a regular alternation of tones of 2 pitches where either of the 2 tones infrequently repeated (P = 0.20), and (4) a random presentation of tones of 5 different pitches, where any of the 5 tones infrequently repeated (P = 0.20). In the count conditions, the infrequent events were designated as targets. It was found that the MMN was elicited by stimulus change and not stimulus repetition in the ignore and count conditions, whereas the N2b was elicited by both stimulus changes and repetitions in the count conditions. It was also possible, in the count conditions, to disentangle the part of the late positive complex which is related to stimulus deviation and the part which is related to stimulus significance (target).


On the evidence of auditory evoked magnetic fields as an objective measure of tinnitus. The purpose of the present study was to determine the utility of auditory evoked magnetic fields as an objective measure of tinnitus. The auditory evoked magnetic fields of 14 patients with tinnitus and of 14 sex- and age-matched controls were measured by means of a 7-channel BTI neuromagnetometer. Stimuli were 1 kHz tone-bursts presented randomly. Tinnitus patients and controls were similar with respect to latencies and amplitudes of the N100m and P200m, and with respect to the locations and moments of the equivalent current dipoles. The present study could not support the notion that specific abnormalities of the auditory evoked magnetic fields are characteristic of patients with tinnitus.


Modelling magnetic coil excitation of human cerebral cortex with a peripheral nerve immersed in a brain-shaped volume conductor: the significance of fiber bending in excitation. To help elucidate some basic principles of magnetic coil (MC) excitation of cerebral cortex, a model system was devised in which mammalian phrenic nerve, or amphibian sciatic nerve with its branches was suspended in appropriate Ringer's solution in a human brain-shaped volume conductor, an inverted plastic skull. The nerve was recorded monophasically out of the volume conductor. The site of nerve excitation by the MC was identified by finding where along the nerve a bipolar electrical stimulus yielded a similar action potential latency. MC excitation of hand-related corticospinal (CT) neurons was modelled by giving the distal end of nerve attached to the lateral skull an initial radial (perpendicular) trajectory, with subsequent bends towards the base and posterior part of the skull; this nerve was optimally excited by a laterally placed figure 8 or round MC when the induced electric field led to outward membrane current at the initial bend. By contrast, nerve given a trajectory modelling CT neurons related to the foot was optimally excited when the coil windings were across the midline, but again when membrane current flowed outward at the first bend. Corticocortical fibers were modelled by placing the nerve in the anteroposterior axis lateral to the midline; with the round MC vertex-tangentially orientated, optimal excitation occurred at the bend nearest the interaural line, i.e., near the peak electric field. The findings emphasize the importance of orientation and direction of current in the MC and fiber bends in determining nerve excitation. The findings in the peripheral nerve-skull model help explain (1) why lateral and vertex-tangentially orientated MCs preferentially excite arm-related CT neurons directly and indirectly (through corticocortical fibers), respectively, and (2) why the MC orientations for optimally exciting directly arm and leg-related CT neurons differ.


Facilitation of motor evoked potentials by somatosensory afferent stimulation. The effect of an electrically induced peripheral afferent volley upon electrical and magnetic motor evoked potentials (MEPs) from muscles of the upper and lower extremities was studied in 16 healthy volunteers. A standard conditioning-test (C-T) paradigm was employed whereby the test stimulus (transcranial electric or magnetic) was applied at random time intervals, from 10 msec prior to 90 msec after the conditioning stimulus (peripheral nerve stimulus). MEP amplitude facilitation was observed for the majority of the upper extremity muscles tested at two distinct periods, one occurring at short, and the other at long C-T intervals. This bimodal trend of MEP facilitation was found to be equally as prominent in the lower extremity muscles tested. The period of short C-T interval facilitation is consistent with modifications in the spinal excitability of the segmental motoneuron pool. On the other hand, the period of long C-T interval facilitation is suggested to be due to alterations in excitability of the motor cortex as a result of the arrival of the orthodromic sensory volley. Although most pronounced in muscles innervated by the nerve to which the conditioning stimulus was applied, this bimodal facilitatory effect was also observed in adjacent muscles not innervated by the stimulated nerve. Qualitatively, the conditioned MEPs from the upper and lower extremities responded similarly to both electrical and magnetic trans-cranial stimulation. In addition, our study demonstrates that the C-T paradigm has potential for use in the assessment of spinal and cortical sensorimotor integration by providing quantitative information which cannot be obtained through isolated assessment of sensory and/or motor pathways.(ABSTRACT TRUNCATED AT 250 WORDS)


The contribution of magnetic stimulation of the motor cortex to the diagnosis of cervical spondylotic myelopathy. Correlation of central motor conduction to distal and proximal upper limb muscles with clinical and MRI findings. Magnetic stimulation of the motor cortex and cervical spine was performed on 24 patients with cervical spondylotic myelopathy documented by MRI. Compound motor action potentials (CMAPs) were recorded from the biceps and thenar muscles to study the central motor pathways of two different myotomes, C5-C6 and C8-D1. Central motor conduction was abnormal in all 24 patients for thenar muscles and in 5 patients for biceps brachii. In patients with a single compression level, central motor conduction abnormalities were confined to the myotomes caudal to the site of compression documented by MRI, in both proximal and distal upper limb muscles in the patients with upper spondylotic compression, and in distal muscles only in the patients with lower compression. In the patients with multilevel compression, central motor conduction time was abnormal for thenar muscles and always normal for the biceps muscle, but its mean value was significantly greater than in the control subjects, suggesting a slight involvement of central motor pathways for proximal upper limb muscles and major damage of the lower cervical segments. Owing to their high degree of sensitivity, central motor conduction studies may be of considerable value in the functional assessment of central motor pathways in cervical spondylotic myelopathy.


The relation between functional deficits, motor and sensory conduction times and MRI findings in syringomyelia. Opinions about the relation between the dimensions of the syrinx and the severity and distribution of symptoms in patients with syringomyelia are controversial. Therefore, this study investigates the relation of clinical symptoms, a disability score, quantified (1- and 2-dimensional) radiological findings (magnetic resonance imaging, MRI) and electrophysiological data (somatosensory and motor evoked potentials) in 22 patients with syringomyelia. There was a close relation between clinical symptoms and electrophysiological data. By both electrophysiological methods subclinical deficits could be detected. Furthermore, the results disclosed that the clinical symptoms, the degree of disability and the duration of the disease are not related to the dimensions of the syrinx or the electrophysiological results obtained by investigating the long ascending and descending spinal tracts of the lower limbs. Our findings suggest that, besides the syrinx, other factors not directly visible in the MRI are responsible for the development and progress of clinical symptoms.


The influence of the shape of mechanical stimuli on muscle stretch reflexes and SEP. Stretching of human thenar muscle was carried out using mechanical stimuli of different amplitudes and velocities in 50 normal subjects. Amplitudes of early M1 and late M2 reflex responses as well as cortical evoked potentials were recorded. M1 and M2 increased with a larger mechanical stimulus. Steeper mechanical stimuli generated larger M1 amplitudes, whereas M2 remained unaffected. Amplitudes of early cortical evoked potentials increased with increase of both rise-time and amplitude of muscle stretch. The shape of the mechanical stimulus is critical for the resulting reflex response. The different behaviours of M1 and M2 with increasing stimulus velocity are in favour of their different origins. Our results support a participation of slowly conducting muscle spindle afferents in the generation of the late M2 response in distal human hand muscles.


Recovery functions of common peroneal, posterior tibial and sural nerve somatosensory evoked potentials. We studied recovery functions of the somatosensory evoked potentials (SEPs) of common peroneal (CPN), posterior tibial (PTN) and sural nerves (SN) using a paired conditioning-test paradigm. The interstimulus interval (ISI) of paired stimuli ranged from 2 to 400 msec. In all SEPs with ISIs of 12-20 msec, the amplitude recovery was close to or beyond 100% of the control response, though their latencies and wave forms were not the same as the control. Further increases of the ISI resulted in significant depression of SEP (late phase suppression), most markedly in CPN, and less prominently in SN-SEP. With a longer than 50 msec ISI there was progressive recovery of SEP, but full recovery differed depending on the nerve stimulated; 400 msec ISI was required for CPN-, 250 msec for PTN- and 100 msec for SN-SEP. The peroneal nerve block by local anesthetic injected just distal to the stimulus electrodes abolished the late phase SEP suppression observed before the nerve block. These findings suggest that the late phase SEP suppression is attributable to the "secondary" afferents as a result of activation of peripheral receptors (muscle, joint and/or cutaneous) by the efferent volley initiated from the stimulus point. The greater and longer duration of peripheral receptor activation in CPN than in PTN or SN stimulation could explain the more pronounced and the longer duration of late phase suppression in CPN-SEP.


After-potentials and control of repetitive firing in human motoneurones. Characteristics of motoneurone after-potentials in man were derived from the recovery curve of motoneurone excitability after a single discharge evoked by threshold stimulation of Ia afferents or by gentle voluntary muscle contraction. The motoneurone excitability was estimated by the firing index of a single motor unit whose potentials were recorded by needle electrodes. The soleus (a slow muscle) and the flexor carpi ulnaris (a fast muscle) were investigated. The duration of motoneurone after-hyperpolarization of the soleus evaluated by this method ranged between 145 and 255 msec; for the flexor carpi ulnaris it was 55-150 msec. In some motoneurones of the fast muscle, an early short-lasting recovery of excitability (within 5-20 msec after a discharge) was revealed. It was accounted for by delayed depolarization of the motoneurone. The relationship between after-potentials and the characteristics of repetitive firing of motoneurones activated by weak voluntary muscle contraction was analysed. It was observed that the motoneurones with early excitability recovery were capable of firing double discharges with a 5-15 msec interspike interval. It was found also that the minimal firing rate of motoneurones (up to 3.1-5.2 imp/sec in the soleus and 3.8-9.0 imp/sec in the flexor carpi ulnaris) was not correlated with the after-hyperpolarization duration. This differs from the results obtained for cat's motoneurones under intracellular stimulation. The findings suggest that after-hyperpolarization is not the only leading mechanism controlling the low firing rate of motoneurones under conditions of their natural activity in man.


Evidence for a novel insulin-like growth factor (IGF)-dependent protease regulating IGF-binding protein-4 in dermal fibroblasts. The mechanisms by which insulin-like growth factors (IGFs) reduce IGF-binding protein-4 (IGFBP-4) levels in cellular conditioned media are poorly understood. The effect of IGFs on IGFBP-4 levels in fibroblast conditioned media is not mediated via the type 1 or type 2 cellular IGF receptors, and the IGFs exert little or no effects on IGFBP-4 messenger RNA levels in human adult fibroblasts or in rat neuroblastoma cells. To determine whether the effects of IGFs on IGFBP-4 might be exerted through alterations in IGFBP-4 degradation, we incubated cell-free, fibroblast-conditioned media from either sheep or human dermal fibroblasts with or without IGF-I, IGF-II (each 1 microgram/ml), or insulin (10 micrograms/ml) for 72 h at 37 C. Samples were then analyzed by Western ligand blot using radiolabeled IGFs and by immunoblotting using a polyclonal antisera to human IGFBP-4. In the absence of IGFs, no apparent changes in the basal concentrations of the various IGFBPs were observed. In contrast, incubation of media with IGFs caused a 70-80% reduction in levels of both sheep and human IGFBP-4, whereas incubation with insulin was without effect. Similarly, incubation of cell-free conditioned media containing recombinant human IGFBP-4 with IGF-I caused a reduction in detectable levels of the 28K protein. The decrease in IGFBP-4 levels was accompanied by the appearance of an immunoreactive approximate 17-20K fragment that did not bind radiolabeled IGFs by ligand blot. The IGF-dependent decrease in IGFBP-4 was prevented by coincubation of the media with serine protease inhibitors, EDTA, or 1,10-phenanthrolene, suggesting that IGFs may activate an IGFBP-4 specific metallo-serine protease present in fibroblast conditioned media. Alternatively, binding of IGF-I or -II to IGFBP-4 may enhance the susceptibility of IGFBP-4 to proteolytic degradation. The demonstration that IGF-I and IGF-II can promote directly the proteolytic degradation of IGFBP-4 into fragments that do not bind IGFs provides a novel mechanism by which the IGFs may increase their own availability and/or activity in biological fluids.


Regulation of galanin gene expression in the rat anterior pituitary gland by the somatostatin analog SMS 201-995. In addition to inducing pituitary tumors in rats, estrogen (E2) markedly increases galanin and PRL gene expression. We previously showed that galanin secretion from pituitary cells in vitro is inhibited by dopamine and somatostatin and stimulated by TRH. The objectives of these in vivo studies were to assess whether the long-acting somatostatin analog SMS 201-995 alters 1) immunoreactive galanin or PRL levels in the anterior pituitary, neurointermediate lobe, hypothalamus, or plasma, 2) pituitary galanin and PRL mRNA levels, and 3) the development of E2-induced pituitary tumors. Ovariectomized Fischer 344 rats were implanted with E2-filled or empty Silastic capsules and treated with or without SMS 201-995 (1.5 mg) via Alzet miniosmotic pumps. Two or 6 weeks later, immunoreactive galanin and PRL levels were determined by RIA. In ovariectomized rats, the somatostatin analog lowered the anterior pituitary content of galanin by 50%, but had no effect on PRL concentrations. E2 increased galanin and PRL levels in the anterior pituitary by 220- and 4-fold, respectively. Concomitant E2 and SMS 201-995 treatment further increased galanin and PRL in the anterior pituitary by 60-80%, but decreased plasma galanin and PRL levels. Likewise, the administration of SMS 201-995 for 2 and 6 weeks inhibited the E2-induced growth of the anterior pituitary. Galanin and PRL mRNA levels were quantified by solution hybridization. Galanin mRNA levels were reduced to undetectable levels in ovariectomized rats treated with SMS 201-995. Furthermore, a 10-fold increase in galanin mRNA levels seen in the presence of E2 was inhibited 80% by SMS 201-995. PRL mRNA levels in E2-treated rats were unchanged by SMS 201-995. We conclude that SMS 201-995 1) lowers plasma galanin and PRL levels in E2-treated rats, 2) elevates the anterior pituitary contents of galanin and PRL in E2-exposed rats, probably through decreased secretion of the hormones, and 3) reduces galanin mRNA levels in E2-treated and untreated ovariectomized rats. Overall, these results establish the differential regulation of galanin and PRL gene expression in vivo by SMS 201-995. Moreover, the data demonstrate that somatostatin receptor agonists may have therapeutic potential for some prolactinomas.


Components of signaling pathways for insulin and insulin-like growth factor-I in muscle myoblasts and myotubes. To survey and compare the signaling pathways from the insulin and insulin-like growth factor-I (IGF-I) receptors in undifferentiated and differentiated muscle cells, we examined the phosphotyrosine (Ptyr)-containing polypeptides elicited in L6 and Sol8 myoblasts and myotubes by the combination of insulin and IGF-I. These polypeptides were detected by immunoblotting with antibodies against Ptyr. In the L6 myoblasts and myotubes and the Sol8 myoblasts, Ptyr polypeptides of approximately 240, 175, 115, 100, 41, and 37 kilodaltons (kDa) appeared in response to insulin-IGF-I. With the Sol8 myotubes, the 240-, 175-, and 37-kDa Ptyr polypeptides were detected in basal cells, and only the Ptyr content of the 175-kDa one increased in response to insulin-IGF-I. The polypeptides of 175, 41, and 37 kDa were tentatively identified as the insulin receptor substrate 1 (IRS1) and extracellular signal-regulated kinases 1 and 2 (ERK1 and -2), respectively, by immunoblotting with antibodies specific for these proteins, and the 115- and 100-kDa polypeptides are probably the beta-subunits of the insulin and IGF-I receptors. The amounts of IRS1, ERK1, and ERK2 were roughly the same in the L6 and Sol8 myoblasts and myotubes. Thus, differentiation of the myoblasts to myotubes was not accompanied by the detectable appearance of new insulin-IGF-I-elicited Ptyr polypeptides or marked changes in the amounts of known participants in their signaling pathways.


Basic fibroblast growth factor modulates insulin-like growth factor-I, its receptor, and its binding proteins in hypothalamic cell cultures. Interactions between different growth factors may be important in the regulation of cell growth and differentiation in the nervous system. For instance, basic fibroblast growth factor (bFGF) regulates neuroblast division through a mechanism probably involving insulin-like growth factor-I (IGF-I). In this regard, we previously found that simultaneous addition of both factors produces an additive effect on survival and differentiation of hypothalamic neuronal and glial cells in culture. To further analyze these interactions, we explored the influence of bFGF on IGF-I, its membrane receptor, and its binding proteins in hypothalamic cells. We also tested the effects of IGF-I on its own receptor and binding proteins (IGFBPs) to determine the specificity of bFGF's actions. Treatment of neuronal and glial cultures with bFGF produced an increase in IGF-I receptors, without changing their affinity, together with an increase in the apparent M(r) of the receptor. On the other hand, IGF-I elicited a down-regulation of its own receptor in both neurons and glia, without modifying its affinity. Treatment with bFGF also produced a marked differential effect on the IGFBPs secreted by the cells. While IGFBP levels in neuronal cultures were greatly increased by bFGF, their production by glial cells was inhibited. On the other hand, IGF-I increased the amount of IGFBPs in both types of cells. Finally, addition of bFGF to the cultures elicited a dose-dependent increase in the release of IGF-I to the medium, but only a moderate increase in cellular IGF-I content, in both neurons and glia. We conclude that bFGF strongly modulates IGF-I, its receptors, and its binding proteins in the two major cell types of the hypothalamus. These findings reinforce the possibility that IGF-I and/or its receptors and binding proteins are involved in the trophic effects of bFGF on developing brain cells.


Modulation of growth-related gene expression and growth inhibition by cyclic adenosine 3',5'-monophosphate-elevating agents in the insulin-producing cell line beta TC1. We studied the involvement of the cAMP pathway in the regulation of beta TC1 cell growth with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) and the activator of adenylate cyclase forskolin. We examined the effect of the increase in cAMP content on the serum-induced resumption of the cell cycle of quiescent cells. IBMX and forskolin both inhibited the mitogenic effect of serum in a concentration-dependent manner. Intracellular cAMP levels were, respectively, enhanced 3.0- and 8.6-fold by IBMX (0.5 mM) and forskolin (20 microM) within 1 h. IBMX and forskolin were also inducers of insulin release, indicating that the growth-arrested beta TC1 cells have retained the essential characteristics of the normal differentiated beta-cells. The effects of IBMX and forskolin were correlated with a modulation of cell cycle-related gene expression. IBMX induced expression of the c-fos gene, which was further enhanced by the simultaneous addition of serum, whereas forskolin alone elicited maximal induction of this gene. Interestingly, c-jun expression was only enhanced with forskolin. We also studied the effects of IBMX and forskolin on the expression of the simian virus-40 T-antigen controlled by the rat insulin II promoter in beta TC1 cells. IBMX and forskolin inhibited the serum-induced accumulation of simian virus-40 T-antigen mRNA in quiescent as well as exponentially growing beta TC1 cells.


Regulation of rat insulin-like growth factor-binding protein-1: the effect of insulin-induced hypoglycemia. Rat insulin-like growth factor-binding protein-1 (rIGFBP-1) was purified from H4IIE rat hepatoma cells by IGF-I affinity chromatography and reverse-phase HPLC. A rabbit antiserum (B2) was raised to rIGFBP-1 and a RIA established. Immunoreactive IGFBP-1 was present in rat amniotic fluid and in the medium conditioned by isolated rat hepatocytes and HTC rat hepatoma cells. To study the effect of hypoglycemia, fasting female Wistar rats were anesthetized and cannulated for multiple venous sampling after the administration of insulin or saline. Serum IGFBP-1 rose in adrenal intact rats from < 0.1 micrograms/ml to a maximum of 1.41 +/- 0.23 micrograms/ml approximately 120 min after insulin administration. Compared to adrenal-intact rats, adrenalectomized animals demonstrated a delayed rIGFBP-1 response to hypoglycemia and did not appear to have reached a maximum at 180 min. A slow rise in rIGFBP-1 levels throughout the sampling period was seen after saline injection in both adrenal-intact and adrenalectomized animals. Glucose, corticosterone, rat insulin, and human insulin levels were measured and none, alone, appeared responsible for the observed rIGFBP-1 responses. We conclude that 1) rIGFBP-1 is stimulated in response to hypoglycemia in a similar manner to glucose counterregulatory hormones, 2) an adrenal factor is required for an early rIGFBP-1 response to hypoglycemia, and 3) neither circulating glucose nor insulin levels, alone, are responsible for the observed patterns of response.


In vitro androgenic induction of a major protein in epithelial cell subcultures from mouse vas deferens. Pure epithelial cell cultures, obtained from primary culture of vas deferens tissue collected from 20- to 30-day-old mice, were amplified by subculturing the cells over 3T3 feeder layer in a serum-free defined medium. Adhesion and proliferation of epithelial cells did not require androgens, but a minimal concentration of 5.10(-7) M hydrocortisone. In that system, epithelial cells expressed cytokeratin but failed to produce the tissue specific mouse vas deferens protein (MVDP) in response to androgens. Various culture procedures and medium compositions were assayed for induction of MVDP expression. Culture onto microporous membrane inserts, which allow polarization of cells, is absolutely required for androgenic induction of MVDP. Androgen action did not require the presence of hydrocortisone, insulin, triiodothyronine, pituitary extracts, epidermal growth factor and acetylcholine. A minimal supplemented medium was then defined in which the expression of MVDP by epithelial cells in response to androgens was dose dependent. It has also been shown that this response at each concentration of dihydrotestosterone was heterogeneous at individual cell level. Highly reproducible results were obtained from epithelial cell cultures between 8th to 16th passages, showing that subcultured cells have maintained their ability to differentiate and express specialized functions.


Somatostatin analog induces insulin-like growth factor binding protein-1 (IGFBP-1) expression in human hepatoma cells. As the somatostatin analog octreotide suppresses pituitary GH secretion and circulating IGF-1 levels, we examined its effects on human hepatoma (hep G2) cells which selectively express IGFBP-1. Octreotide (60 nM) stimulated IGFBP-1 up to 4.1-fold (p < 0.001 after 24 hrs). Induction of IGFBP-1 was first detectable after 12 hrs of 6 nM octreotide (1.5-fold, p < 0.03), and was confirmed by ligand blotting. Cholera toxin and forskolin induced IGFBP-1 independently and were also additive with octreotide. IGFBP-1 mRNA expression was induced 2.7-fold by octreotide. Thus, octreotide induces basal and stimulated IGFBP-1 in hepatocytes independently of insulin and GH. As IGFBP-1 may regulate peripheral IGF-1 action, induction of IGFBP-1 represents a novel pituitary-independent mechanism for octreotide action.


Prorenin-renin axis in synovial fluid in patients with rheumatoid arthritis and osteoarthritis. This study was undertaken 1) to determine whether or not renin is present in synovial fluid in patients with rheumatoid arthritis and osteoarthritis, and, if present, 2) to investigate whether it is synthesized in synovial fluid, or it is only transported from the circulation into the synovial cavity. The active renin concentration (indirect) was measured with angiotensin I radioimmunoassay kits. Inactive renin was converted into active renin with Sepharose-bound trypsin. Both active and inactive forms of renin were found in synovial fluid. They were significantly higher in patients with rheumatoid arthritis (n = 9) than in those with osteoarthritis (n = 16). In plasma, the concentration of inactive renin was significantly higher (P less than 0.001) in the former. Albumin, transferrin, alpha 2-macroglobulin, ceruloplasmin and immunoglobulins G and M were also found in synovial fluid. In each disease, a plot of the log ratio of synovial fluid to the serum concentration against the log molecular weight of each protein gave an approximately straight line curve, suggesting that these proteins are derived from the circulation and are transported into the synovial cavity. In contrast, the ratio of synovial fluid to plasma concentrations of active renin was significantly higher than that predicted on the basis of the above-mentioned interrelationships in both diseases, whereas the ratio of inactive renin was significantly lower. These findings suggest that 1) inactive and active renin are filtered into the synovial fluid from the circulation, and that 2) inactive renin is converted into the active form in the fluid.


Localized non-Hodgkin's lymphoma of the adrenal and thyroid glands. A case of immunoblastic lymphoma, involving only the thyroid and the adrenal glands, is presented. The patient had clinical symptoms and findings of Addison's disease, and computed tomography (CT) demonstrated bilateral adrenal tumoral enlargement. He also had euthyroid diffuse multinodular goiter. The diagnosis of the patient was based on the cytological examination of the aspiration materials from both endocrine glands. The patient received "m-BNCOD" chemotherapy regimen and replacement therapy for Addison's disease. At the end of three courses, a partial response was obtained.


The orientation of transition moments of dye molecules used in fluorescence studies of muscle systems. Fluorescence and phosphorescence depolarization techniques can provide information on orientational order and rotational motion of crossbridges in muscle fibres. However the depolarization experiment monitors the orientation and motion of the crossbridges indirectly. The changes in depolarization arise from a change in the orientation of the transition dipoles of the dye attached to the crossbridge. In order to extract the physiologically relevant orientations from the data it is therefore necessary to characterize the orientation of the dye molecule relative to the crossbridge and the orientation of the transition moments in the frame of the dyes. The dyes 1,5-I-AEDANS and eosin-5-maleimide are commonly used for labelling the crossbridge in muscle fibres. The orientations of the absorption and fluorescence emission dipoles of these two dyes in the molecular frame were determined. Angle resolved fluorescence depolarization experiments on the dyes, macroscopically aligned in a stretched polymer matrix of poly vinyl alcohol, were carried out. The data were analyzed in terms of an orientational distribution of the dye molecules in the film and the orientations of the absorption and emission dipoles in the frame of the dye molecule. Experimental data, obtained from a given sample at different excitation wavelengths, were analyzed simultaneously in a global target approach. This leads to a reduction in the number of independent parameters optimized by the non-linear least squares procedure.


The roles of serine and threonine sidechains in ion channels: a modelling study. The ion channel of the nicotinic acetylcholine receptor (nAChR) is believed to be lined by transmembrane M2 helices. A "4-8-12" sequence motif, comprising serine (S) or threonine (T) residues at positions 4, 8 and 12 of M2, is conserved between different members, anion and cation selective, of the nAChR superfamily. Parallel bundles of 4-8-12 motif-containing helices are considered as simplified models of ion channels. The relationship between S and T sidechain conformations and channel-ion interactions is explored via evaluation of interaction energies of K+ and of Cl- ions with channel models. Energy calculations are used to determine optimal chi 2 (C alpha-C beta-O gamma-H gamma) values in the presence of K+ or Cl- ions. 4-8-12 motif-containing bundles may form favourable interactions with either cations or anions, dependent upon the chi 2 values adopted. Parallel-helix and tilted-helix bundles are considered, as are heteromeric models designed to mimic the Torpedo nAChR. The main conclusion of the study is that conformational flexibility at chi 2 enables both S and T residues to form favourable interactions with anions or cations. Consequently, there is apparently no difference between S and T residues in their interactions with permeant ions, which suggests that the presence of T vs. S residues within the 4-8-12 motif is not a major mechanism whereby anion/cation selectivity may be generated. The implications of these studies with respect to more elaborate models of nAChR and related receptors are considered.


Using coomassie blue to stabilize H2O2-guaiacol stained peroxidases on polyacrylamide gels. Shimoni and Reuveni [1] have reported a procedure which uses Coomassie Brilliant Blue R-250 staining and destaining to stabilize H2O2-guaiacol stained peroxidases on polyacrylamide gels and at the same time detects nonspecific protein bands. The procedure has the advantage that the otherwise orange and unstable peroxidase bands are visualized as blue bands and it, therefore, facilitates laser densitometry of electrophoretically separated peroxidases. In the modification of the procedure reported here, the Coomassie staining reagent was adjusted and the destaining step was eliminated. The modified procedure has added advantages: it simplifies the procedure, increases the consistency of the results across gels, prevents or reduces the staining of nonspecific proteins, and still gives excellent resolution of stable bands which can be used to quantify peroxidase activity.


Fibroblast growth factor receptor-4 shows novel features in genomic structure, ligand binding and signal transduction. Fibroblast growth factor (FGF) receptor (FGFR) gene family consists of at least four receptor tyrosine kinases that transduce signals important in a variety of developmental and physiological processes related to cell growth and differentiation. Here we have characterized the binding of different FGFs to FGFR-4. Our results establish an FGF binding profile for FGFR-4 with aFGF having the highest affinity, followed by K-FGF/hst-1 and bFGF. In addition, FGF-6 was found to bind to FGFR-4 in ligand competition experiments. Interestingly, the FGFR-4 gene was found to encode only the prototype receptor in a region where both FGFR-1 and FGFR-2 show alternative splicing leading to differences in their ligand binding specificities and to secreted forms of these receptors. Ligands binding to FGFR-4 induced receptor autophosphorylation and phosphorylation of a set of cellular polypeptides, which differed from those phosphorylated in FGFR-1-expressing cells. Specifically, the FGFR-1-expressing cells showed a considerably more extensive tyrosine phosphorylation of PLC-gamma than the FGFR-4-expressing cells. Structural and functional specificity within the FGFR family exemplified by FGFR-4 may help to explain how FGFs perform their diverse functions.


The multidrug resistance and cystic fibrosis genes have complementary patterns of epithelial expression. The cystic fibrosis gene product, CFTR, and the multidrug resistance P-glycoprotein (encoded by the MDR1 gene) are structurally related proteins and both are associated with epithelial chloride channel activities. We have compared their cell-specific expression in the rat by in situ hybridization. In all tissues examined the two genes were found to have complementary patterns of expression, demonstrating exquisite regulation in both cell-specific and temporal fashions. Additionally, a switch in expression from one gene to the other was observed in certain tissues. For example, expression in the intestine switches from CFTR to MDR1 as the cells migrate across the crypt-villus boundary. A switch from CFTR to MDR1 expression was also observed in the uterine epithelium upon pregnancy. These data suggest that CFTR and P-glycoprotein serve analogous roles in epithelial cells and provide additional evidence that P-glycoprotein has a physiological role in regulating epithelial cell volume. The patterns of expression suggest that the regulation of these two genes is coordinately controlled.


The atypical M2 segment of the beta subunit confers picrotoxinin resistance to inhibitory glycine receptor channels. Purified preparations of the inhibitory glycine receptor (GlyR) contain alpha and beta subunits, which share homologous primary structures and a common transmembrane topology with other members of the ligand-gated ion channel superfamily. Here, a beta subunit-specific antiserum was shown to precipitate the [3H]strychnine binding sites localized on alpha subunits from membrane extracts of both rat spinal cord and mammalian cells co-transfected with alpha and beta cDNAs. Further, inhibition of alpha homo-oligomeric GlyRs by picrotoxinin, a non-competitive blocker of ion flow, was reduced 50- to 200-fold for alpha/beta hetero-oligomeric receptors generated by cotransfection. Site-directed mutagenesis identified residues within the second predicted transmembrane segment (M2) of the beta subunit as major determinants of picrotoxinin resistance. These data implicate the M2 segment in blocker binding to and lining of the GlyR chloride channel.


The human T cell antigen gp39, a member of the TNF gene family, is a ligand for the CD40 receptor: expression of a soluble form of gp39 with B cell co-stimulatory activity. Signals delivered to B cells via CD40 can synergize with those provided by other B cell surface receptors to induce B cell proliferation and antibody class switching as well as modulate cytokine production and cell adhesion. Recently, it has been shown that the ligand for CD40 is a cell surface protein of approximately 39 kDa expressed by activated T cells, gp39. Here we report on the isolation and characterization of a cDNA clone encoding human gp39, a type II membrane protein with homology to TNF, and the construction and characterization of a soluble recombinant form of gp39. COS cell transfectants expressing gp39 synergized with either anti-CD20 mAb or PMA to drive strong B cell proliferation and alone were able to drive B cells to proliferate weakly. In all cases the B cell proliferation induced by gp39-expressing COS cells was reduced to background levels by the addition of soluble CD40. Unlike gp39-expressing COS cells, recombinant soluble gp39 was not mitogenic alone and required co-stimulation to drive B cell proliferation. These results suggest that B cells require a second signal besides gp39-CD40 to drive proliferation and that soluble gp39 alone in a non-membrane bound form is able to provide co-stimulatory signals to B cells.


Cytokine effects of CD23 are mediated by an epitope distinct from the IgE binding site. Human CD23 and its soluble forms (sCD23) display various biological activities, in addition to their IgE binding function (IgE/BF). The IgE binding domain was recently mapped to residues between Cys163 and Cys282 but its involvement in IgE-independent, CD23 functions remains unknown. In order to clarify this point, a series of N-terminal, C-terminal and internal deletion mutants of CD23 or sCD23 were expressed in CHO cells and tested for their ability (i) to bind to IgE, (ii) to induce colony formation by human myeloid precursor cells, (iii) to promote mature T cell marker expression by early prothymocytes, and (iv) to regulate IgE synthesis. The present study indicates that cytokine activities require the presence of Cys288, while this amino acid is not necessary for IgE/BF. Blocking experiments using various conformation-sensitive monoclonal antibodies further suggest that active epitope(s) of CD23 in cytokine assays is(are) distinct from those involved in IgE/BF.


Identification of two structurally related proteins involved in proteolytic processing of precursors targeted to the chloroplast. Two proteins of 145 and 143 kDa were identified in pea which co-purify with a chloroplast processing activity that cleaves the precursor for the major light-harvesting chlorophyll binding protein (preLHCP). Antiserum generated against the 145/143 kDa doublet recognizes only these two polypeptides in a chloroplast soluble extract. In immunodepletion experiments the antiserum removed the doublet, and there was a concomitant loss of cleavage of preLHCP as well as of precursors for the small subunit of Rubisco and the acyl carrier protein. The 145 and 143 kDa proteins co-eluted in parallel with the peak of processing activity during all fractionation procedures, but they were not detectable as a homo- or heterodimeric complex. The 145 and 143 kDa proteins were used separately to affinity purify immunoglobulins; each preparation recognized both polypeptides, indicating that they are antigenically related. Wheat chloroplasts contain a soluble species similar in size to the 145/143 kDa doublet.


GATA-1 but not SCL induces megakaryocytic differentiation in an early myeloid line. GATA-1, a transcription factor of the 'zinc-finger' family, is required for the development of mature erythroid cells and is also highly expressed in the megakaryocytic and mast cell lineages. The helix-loop-helix gene SCL (or TAL) is expressed in the same three hematopoietic lineages as GATA-1. To explore the role of GATA-1 and SCL in hematopoietic differentiation, we introduced a new expression vector bearing each gene into the early myeloid cell line 416B, which could originally differentiate in vivo along the megakaryocytic and granulocytic lineages. Enforced expression of SCL at high levels did not provoke differentiation, but GATA-1 induced the appearance of megakaryocytes as assessed by morphology, the presence of acetylcholinesterase and a polyploid DNA content. Although GATA-1 is thought to stimulate its own transcription in erythrocytes, expression of the endogenous gene was not increased in the megakaryocytic lines; hence GATA-1 may not be autoregulatory in this lineage. Megakaryocytic differentiation was accompanied by a marked decrease in the myeloid surface marker Mac-1. The absence of mast cell or erythroid differentiation suggests that GATA-1 may not be sufficient to provoke maturation along these lineages or that these pathways are impeded in 416B cells. These results demonstrate that a member of the GATA gene family can act as an important regulator of megakaryocytic differentiation.


The effect of pedaling frequency on glycogen depletion rates in type I and type II quadriceps muscle fibers during submaximal cycling exercise. This study was conducted to determine whether the pedaling frequency of cycling at a constant metabolic cost contributes to the pattern of fiber-type glycogen depletion. On 2 separate days, eight men cycled for 30 min at approximately 85% of individual aerobic capacity at pedaling frequencies of either 50 or 100 rev.min-1. Muscle biopsy samples (vastus lateralis) were taken immediately prior to and after exercise. Individual fibers were classified as type I (slow twitch), or type II (fast twitch), using a myosin adenosine triphosphatase stain, and their glycogen content immediately prior to and after exercise quantified via microphotometry of periodic acid-Schiff stain. The 30-min exercise bout resulted in a 46% decrease in the mean optical density (D) of type I fibers during the 50 rev.min-1 condition [0.52 (0.07) to 0.28 (0.04) D units; mean (SEM)] which was not different (P > 0.05) from the 35% decrease during the 100 rev.min-1 condition [0.48 (0.04) to 0.31 (0.05) D units]. In contrast, the mean D in type II fibers decreased 49% during the 50 rev.min-1 condition [0.53 (0.06) to 0.27 (0.04) units]. This decrease was greater (P < 0.05) than the 33% decrease observed in the 100 rev.min-1 condition [0.48 (0.04) to 0.32 (0.06) units). In conclusion, cycling at the same metabolic cost at 50 rather than 100 rev.min-1 results in greater type II fiber glycogen depletion. This is attributed to the increased muscle force required to meet the higher resistance per cycle at the lower pedal frequency.(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of the reverse transcriptase of 1731, a Drosophila melanogaster retrotransposon. The nucleotide sequence of 1731, a retrotransposon cloned from the genome of Drosophila melanogaster, reveals a structural similarity with the proviral form of the retroviruses including a pol-like gene containing a putative reverse-transcriptase(RT)-coding sequence. Diverse parts of that sequence were subcloned and expressed in Escherichia coli. It has been demonstrated that the expression of the RT-like sequence, when translated, gives rise to peptides displaying enzyme activity characteristic of a true RT enzyme. In addition, rabbit antisera directed against such recombinant proteins allowed us to detect an immunoreactive protein of around 110 kDa, which was only present in D. melanogaster cell lines, but not in cells derived from Drosophila virilis or Drosophila hydei, whose genomes do not bear the 1731 element. This protein is expected to correspond to a non-processed pol-gene translated product and cosediments with virus-like particles exhibiting RT activity.


Isolation and characterization of alliinase cDNA clones from garlic (Allium sativum L.) and related species. cDNA libraries constructed from poly(A)-rich RNA isolated from Allium sativum (garlic), Allium cepa (onion) and Allium ascalonicum (shallot) were screened for cDNA clones encoding the alliinase using colony hybridization. Sequence analysis of the alliinase cDNA clones from different Alliaceae species revealed a high degree of sequence similarity both at the nucleotide and at the amino acid level. Apparently, the alliinases are translated from mRNA species of approximately 2200 nucleotides. The primary translation products are preproproteins which are converted into the mature alliinases following post-translational modifications. In the case of the garlic alliinase, the mRNA encodes a 486-amino-acid polypeptide with a molecular mass of 55,623 Da. Cleavage of the signal peptide (28 amino acids) results in a preprotein which extends 10 amino acids before the first amino acid of the mature protein of 51,451 Da. Southern-blot analysis of genomic DNA has shown that the alliinases are most probably encoded by a family of closely related genes, which is in good agreement with the sequence heterogeneity found between different alliinase cDNA clones of one species.


Stable expression of chicken type-VI collagen alpha 1, alpha 2 and alpha 3 cDNAs in murine NIH/3T3 cells. As a component of an extensive network of microfibrils interwoven with large collagen fibers and in close contact with cell surfaces, type VI collagen plays an important role in cell-matrix interactions. To investigate the behaviour of chicken type VI collagen chains in heterologous host cells as a means to understanding the pattern of assembly of this collagen, we transfected murine NIH/3T3 cells with cDNAs encoding chicken alpha 1(VI), alpha 2(VI) and alpha 3(VI) chains. Cell lines that constitutively expressed the individual chains were analyzed by metabolic labeling and immunoprecipitation with specific antibodies. No self-association was observed for either alpha 1(VI) or alpha 2(VI) chains which were secreted as monomeric polypeptides. Furthermore, neither the chicken alpha 1(VI) nor alpha 2(VI) chains associated with the endogenous murine chains to form chimeric chicken/murine heterotrimers. In contrast, chimeric chicken/murine heterotrimers were detected in cell lines transfected with chicken alpha 3(VI) cDNA. These chimeric forms appeared to be properly aligned since their triple helices were stable to pepsin digestion. In addition, the chimeric heterotrimers coassembled and gave rise to disulfide-linked type VI collagen molecules.


Tenascin: a modulator of cell growth.

Tenascin: a modulator of cell growth. The large, multidomain extracellular matrix protein tenascin displays a markedly restricted tissue distribution during embryogenesis and remains present only in a few adult tissues. The protein is reexpressed, however, during wound healing and in the stroma of malignant tumours. While a variety of studies have dealt with the important role of tenascin in the development of neural and non-neural tissues, there is growing evidence that tenascin expression may be associated with proliferation of cells lining these tissues. The presence of repeating domains in tenascin similar to those in epidermal growth factor prompted us to investigate the ability of tenascin to modulate the growth of different cell types. Tenascin was actually found to be mitogenic for several cell types. This mitogenic activity, however, appears to be associated with a region in the fibronectin type III domains. The mitogenic mechanism is clearly distinct from pathways used by peptide growth factors such as epidermal growth factor and platelet-derived growth factor, which activate the intrinsic tyrosine kinase activity of their cell-surface receptors. However, we show that this large extracellular matrix molecule is efficiently internalised and may be processed by responding cells.


The major zymogen granule membrane protein GP-2 in the rat pancreas is not involved in granule formation. The major membrane protein of zymogen granules in the rat pancreas is a glycoprotein of 78 kDa (GP-2), which is inserted into the membrane via a glycosyl-phosphatidylinositol (GPI) anchor. GP-2 occurs in both, a membrane-attached and a soluble form. Due to its specific luminal orientation and its quantitative contribution to the zymogen granule membrane, GP-2 has been postulated to play an important role in sorting of digestive enzymes into the granule and in the formation of the granule as a storage organelle. We have tested this hypothesis in the rat pancreas under three different functional conditions, where both the rates of enzyme/isoenzyme synthesis change drastically, and new zymogen granules form at a high rate: a) during prolonged hormonal stimulation of the adult rat pancreas, b) during the differentiation of AR4-2J cells induced by dexamethasone in vitro, and c) during embryonic development and early postnatal life, when gene expression is modulated due to the differentiation program. Both, GP-2 mRNA levels and the rate of GP-2 biosynthesis were quantitated and compared to the immunohistochemical localization of this protein in tissue sections. Under all three functional conditions, significant changes could be demonstrated at the level of digestive enzyme gene expression, but no concomitant modulation of GP-2 expression was observed. GP-2 mRNA is absent from the embryonic pancreas and for the first time is expressed after birth with a significant increase during the period of weaning. Furthermore, GP-2 mRNA and protein levels are not modulated by hormonal stimulation, either in the adult pancreas or in AR4-2J cells in culture. Therefore, we conclude that GP-2, in spite of its quantitative contribution to the zymogen granule membrane, is not involved in enzyme protein sorting or granule formation. Alternative functions for GP-2 are discussed.


Antigenic determinants of T lymphocyte alpha beta receptor and other leukocyte surface proteins as differential markers of skeletal muscle regeneration: detection of spatially and timely restricted patterns by MAM microscopy. Different stages of human muscle regeneration have been identified by multiple antigen-mapping (MAM) microscopy at the level of a novel marker system. This immunofluorescence method allows the selective imaging of numerous antigen signals from cells or tissue sections. It is shown that 2 monoclonal antibodies (mAbs) against the common region of the alpha beta T lymphocyte antigen receptor (alpha beta and alpha TCR chains) and 3 mAbs against the leukocyte surface antigens Leu8, OKM5 and Leu19 recognize regenerating muscle cells at different time points of regeneration in human muscle sections. Paradoxically, these epitopes are not expressed by T lymphocytes or other mononuclear leukocytes invading regenerating muscle. Hence, presence of the corresponding antigens in muscle fibers may exclude their expression by muscle-invasive immune cells suggesting a function in muscle-specific cell-to-cell recognition. Simultaneous localization of these epitopes in Duchenne dystrophy reveals 10 different phenotypes of regenerating and normal infantile muscle cells due to different developmental stages during the myocyte differentiation. In adult muscle (mitochondrial myopathy) segmental muscle fiber necrosis is accompanied by high concentration of alpha beta/alpha TCR epitopes in the intact fiber ends, which are the target sites of myogenesis. The same sites are invaded by OKM5+ endomysial capillary sprouts that terminate at the tip of the alpha beta/alpha TCR reactive fiber ends. These hitherto unrecognized initial events of segmental muscle regeneration seem to be followed by fragmentation of the invasive capillaries into single endothelial cells, which then switch from the OKM5+ Leu19- through the OKM5+ Leu19+ to the OKM5- Leu19+ phenotype. This cell type exhibits the typical features of Leu19+ myogenic stem cells described earlier. The findings may give rise to a concept of muscle repair, in which the alpha beta/alpha TCR-related antigen, concentrated in fiber stumps, might provide positional information for invading endothelial cells. These cells appear to be a source of myogenic stem cells for regeneration.


Characteristics of pancreatic exocrine secretion produced by venom from the Brazilian scorpion, Tityus serrulatus. The influence of venom (TSV) from the Brazilian scorpion, Tityus serrulatus, on exocrine pancreatic secretion was studied in relation to known cholinergic and peptidergic secretagogue activity. Pulse-labeling followed by chase incubation in the presence of secretagogues and various pharmacological agents revealed unique physiological characteristics of TSV in guinea pig pancreatic lobules. Exocytotic discharge of newly synthesized 3H-labeled proteins during a 3-h chase incubation showed a marked increase over basal discharge levels using logarithmic TSV doses of 0.10 to 100 micrograms/ml. This stimulation was comparable to maximal values elicited by carbachol, cholecystokinin-octapeptide (CCK-8) or caerulein and discharge kinetics were similar. TSV-mediated secretion was ATP and calcium dependent and partially inhibited by atropine. Only tetrodotoxin completely blocked TSV stimulation of newly synthesized protein discharge. Both botulinum toxin and curare had no effect on venom stimulation, indicating that TSV interaction with exocrine pancreatic cells occurs postsynaptically. Verapamil, a calcium channel antagonist, produced a moderate inhibition of TSV stimulation. When antagonists to the cholecystokinin (CCK) receptor were incubated with TSV, no change in secretory activity occurred. Therefore, TSV does not bind to CCK receptors and probably operates through its own receptor which may be an ion channel. Additionally, morphological studies in vitro revealed a high level of pancreatic secretory activity as evidenced by dense secretory acinar luminal content, reduction in zymogen granule (ZG) population, and development of exocytotic images.


Serum osteocalcin in monitoring bone metastases in advanced prostatic cancer. Osteocalcin, a K-dependent vitamin protein, was studied in a group of advanced prostatic carcinoma patients to test the usefulness of this marker for diagnosing bone metastases. Osteocalcin levels were above the norm in 22 out of 27 patients with bone metastases, although high levels were not observed in patients without bone metastases. High sensitivity and specificity levels of serum osteocalcin appear to be strongly correlated to metastatic bone involvement and disease relapse after hormone treatment. Although these results must be confirmed on a larger series of patients, this protein appears to be a useful biological marker in prostatic cancer.


Advanced prostate cancer follow-up with prostate-specific antigen, prostatic acid phosphatase, osteocalcin and bone isoenzyme of alkaline phosphatase. We report our experience in the follow-up of 63 patients with advanced prostate adenocarcinoma. We used prostate-specific antigen and prostatic acid phosphatase in 27 patients; in 36 patients we evaluated osteocalcin and bone isoenzyme of alkaline phosphatase, two markers of bone metabolism which seem to be good markers in the follow-up of patients with bone metastases.


Relationship between prostatic acid phosphatase and prostate-specific antigen serum levels and prostatic volume in benign prostate hyperplasia. Pitfall on tumor markers assessment in primary prostatic cancer? Serum levels of prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) were measured in 78 patients with benign prostate hyperplasia and compared with both the gland weight and the glandular component of prostatic tissue. Both PAP and PSA were significantly higher where prostate was heavier; however, we could not find a consistent factor which could correlate weight increase to marker levels. PSA tended to be higher when glandular component was more expressed. From the present findings we conclude that in patients with prostate cancer, PAP and PSA serum levels should be investigated considering also the benign components of prostate gland.


Seasonal variation of prostatic acid phosphate and prostate-specific antigen in patients without prostatic malignancy. The seasonal pattern of prostatic acid phosphatase (PAP) and prostate specific antigen (PSA) in nonmalignant males was investigated. Serum levels were measured in 1,540 men during a 3-year period with radioimmunoassay methods using monoclonal antibody techniques. All of the tested individuals were free of prostatic malignancy. During each of the 3 years, PAP ans PSA showed a rise, especially in spring. The mean PSA level in spring showed a statistically important difference when compared with winter, fall and summer mean levels (p less than 0.05). However, no significant difference of PAP levels was estimated seasonally in the 3 years, which shows that an important marker of prostatic cancer can vary with seasons.


A new marker for early detection and indicator of progression of cancer of the bladder. Preliminary results with Ag-NOR index in 38 cases of superficial bladder cancer. The multiform biology of superficial bladder tumors, both morphologically and evolutively, and the lack of reliable predictors of progression have led the authors to study the Ag-NOR proteins as a new marker of these tumors. It is well known that particularly the low-grade superficial tumors frequently relapse on the same histologic and proliferative module. Their potential of progression is probably present at the time of the first manifestation of the disease or it can show itself along the relapsing evolution with classic modifications translating the cellular dedifferentiation. The NOR index, set up by the authors, has several advantages: firstly, it corresponds to a functional value of normal and neoplastic cells; secondly, it can be used also with paraffin blocks. Another advantage is the semiautomatic lecture, reproducible also in the urinary cytology, mainly of low-grade tumors, reducing the number of false-negatives. The conclusion of the study of 38 cases of superficial bladder cancer has induced the authors to believe that an increased NOR index is a reliable 'marker' of their progression. Therefore, the authors suggest the use of the NOR activity for the surveillance of the urothelial disease and for a more logical therapeutic strategy.


Current status of tumor markers in testicular cancer. A practical review. Detection of serum and cellular AFP and hCG has made a significant contribution in understanding and management of testicular cancer. It is essential to remember the following events in utilizing these markers: (1) Histologic diagnosis of seminoma, but AFP is elevated. There is usually an element of choriocarcinoma. (2) Histologic diagnosis of seminoma and highly elevated hCG greater than 100 ng/ml has usually an element of choriocarcinoma. (3) Histologic diagnosis of choriocarcinoma with an elevated serum AFP. There is usually an element of embryonal carcinoma. (4) Pathologic stage I nonseminomatous testicular cancer with elevated serum markers is either stage II or stage III. (5) In a recent study of 23 patients undergoing resection of residual nonseminomatous testicular cancer after intensive chemotherapy, 21 had either teratoma in primary tumor or bulky metastatic disease. The markers were normalized after chemotherapy and prior to resection. (6) Although normalization of these markers after chemotherapy indicates effective therapeutic response, one should look of residual tumor utilizing radiologic investigations.


Beta-human chorionic gonadotropin and alpha-fetoprotein in central and peripheral venous blood of patients with testicular tumors. From January 1989 to June 1991, 18 patients ranging in age from 19 to 41 (mean 34) years with testicular tumor were examined. 14 patients had seminoma (11 typical and 3 spermatocytic) and 4 patients had a mixed form (2 seminoma + embryonal tumor and 2 seminoma + teratocarcinoma). Serum levels of beta-human chorionic gonadotropin and alpha-fetoprotein from peripheral venous blood and from spermatic venous vessel were evaluated in every patient. All patients with seminoma and in a patient with mixed tumor (seminoma + embryonal tumor) the markers were regular. The increase of the markers was found in the peripheral and in spermatic blood of 3 patients (2 seminoma + embryonal carcinoma and 1 seminoma + teratocarcinoma). For these reasons the values of spermatic vessels are an important confirmation of the level of peripheral markers.


Evaluation of hematochemical parameters and renal echography after ESWL. Evaluation of hematochemical parameters and renal echography after ESWL treatment with Tripter X1 was done on the first 95 patients treated and for whom we had a longer than 3-month follow-up. Creatinine phosphokinase values showed an increase in 24 patients. 12 patients demonstrated an increase in amylasemia. Echographic checks performed the day after treatment revealed the presence of perirenal lesions in 3 cases. As far as amylasemia is concerned, only 1 patient was treated with aprotinin. One of the patients who developed perirenal hematoma had hypertension; another was following an antiaggregant therapy that was temporarily suspended.


Tissue markers in the diagnosis and prognosis of prostatic carcinoma. Prostatic acid phosphatase (PAP), prostate-specific antigen (PSA), and beta-microseminoprotein are organ-specific markers. Also Leu-7 may be regarded as an organ-specific marker. The main utilization of PAP and PSA immunostaining is to establish the prostatic origin of a carcinoma. Changes of PAP and PSA immunoreactivity may be correlated with histologic grade of prostatic carcinoma. Keratin expression assessment is useful to identify prostatic basal cells or epithelial secretory cells. Neuroendocrine marker reactivity in prostatic carcinoma seems to be related with increasing histologic grade and tumor progression.


Correlation between serum values of prostatic acid phosphatase and morphometric analysis in the cytologic diagnosis of prostatic carcinoma. We studied 78 men with suspicion of prostatic carcinoma, who underwent transrectal aspiration biopsy, diagnosing 46 adenocarcinoma, 13 chronic prostatitis and 19 benign prostatic hyperplasia. Moreover, we determined prostatic acid phosphatase (PAP) by enzyme immunoanalysis, resulting in 9/78 false-positives and 18/78 false-negatives. Also, we carried out a morphometric analysis of the cytologic samples which showed good correlation with the cytologic diagnosis except in the moderately differentiated carcinomas. We found a good correlation between PAP values, cytologic diagnosis and nuclear size as well as the percentage of the binucleolated cells.


New ultrasensitive assay development by using monoclonal antibodies for detecting prostate-specific antigen. The Serono Maia Clone prostatic-specific antigen (PSA) kit incorporates an immunoradiometric assay for the measurement of PSA in the serum. The method can be used over a range of 0-100 ng/ml without dilution. Standard concentrations are 0, 0.4, 1, 5, 20, and 100 ng/ml. Up to date, 373 normal men, 89 normal women, 117 prostatic carcinoma, 98 other carcinoma, and 85 benign prostatic hypertrophy have been tested. The aim of this study is to evaluate the sensitivity and specificity of a new immunoassay method for the determination of PSA, that could be able to evaluate low levels of PSA, resulted undetectable with other methods. This ability could be useful in patients treated with hormone-suppressive therapy or after radical prostatectomy. We have collected all low values present in samples examined. With the Serono Maia Clone PSA kit only 26.7% of these have been evaluated as 'out' values as opposed to 46.5% with the Hybritech kit.


Changes in prostate-specific antigen and prostatic acid phosphatase concentration following prostatic examination in benign prostatic hypertrophy and prostate cancer patients. The authors measured serum prostate-specific antigen (PSA) and prostatic acid phosphatase concentration in histologically positive prostate hyperplastic and carcinomatous patients before, and 30-60 min and 24 h after prostate manipulation (rectal digital examination, cystoscopy and perineal punch biopsy). After rectal examination, PSA, and after other interventions, both markers changed significantly, although in different points of time and to a different extent. The authors call the attention to the importance of the point of time of the examination. Examination following prostate manipulation may result in wrong diagnosis or in erroneous therapeutic consequences in the follow-up period.


Comparison between digital rectal examination, prostate-specific antigen and transrectal ultrasound in symptomatic patients. Results on 141 cases. In this study we proposed to verify sensitivity and specificity of prostate-specific antigen (PSA), digital rectal examination (DRE) and transrectal ultrasound (TRUS) in patients who referred at our institution for prostatic complaints. 141 patients, ages ranging between 55 and 86 years (mean 67.5), underwent DRE, blood PSA, TRUS and ultrasonically guided biopsy of the prostate. The comparison of the results obtained with the different diagnostic tools allowed us to draw a diagnostic algorithm for prostate cancer in symptomatic patients.


Prognostic significance of prostate-specific antigen in endocrine treatment for prostatic carcinoma. We have studied the prognostic significance of prostate-specific antigen (PSA), monitored monthly, in 24 patients with prostatic cancer (5 D1, 19 D2) on endocrine therapy. The pretreatment levels of PSA were high in all patients (mean value 41 ng/ml). It was found that PSA levels at the end of the first and sixth months of treatment were reliable prognostic indicators. At the first month evaluation PSA had decreased more than 50% from the initial values in the 16 patients with stable disease, while it had decreased less than 50% in those with progressing disease. At the end of 6 months, patients with stable disease had PSA levels within the normal range, while 8 of the patients who had progressing disease had levels higher than 10 ng/ml. Respectively 6 and 2 patients had also had increases in PSA levels at 3 and 6 months before scintigraphic demonstration of increased bone metastases.


Preoperative and postoperative evaluation of prostate-specific antigen in localized prostatic cancer treated by radical prostatectomy. Preoperative prostate-specific antigen (PSA) values were determined in 73 patients with clinically localized prostatic cancer and candidates for a radical procedure. Correlation of preoperative PSA with a final pathological stage was attempted. Only in 44.8% of our 22 patients with organ-confined disease was the PSA value within the normal range; in 17.3% of cases PSA values were higher than 20 ng/ml. 18.2% of the patients with locally advanced disease showed normal PSA values, while 45.5% had concentrations above 20 ng/ml. In the case of lymph node involvement, PSA values were normal in 22.7% of the cases. Our data indicate that no strict relationship can be suggested between PSA and the final pathological stage and grading of the tumor in patients who underwent radical prostatectomy.


Influence of prostatic disease and prostatic manipulations on the concentration of prostate-specific antigen. We examined the influence of different factors [benign prostatic hyperplasia (BPH), prostatic carcinoma (PCA), organ volume, weight of resected tissue, transurethral catheter] on the serum prostate-specific antigen (PSA) levels in 253 patients with BPH (n = 138; 54%) and PCA (n = 115; 46%). Only in 57.2% of the BPH patients, PSA values were < 4 ng/ml, in 74.6% < 7 ng/ml. In 108 patients with BPH, a transurethral prostatectomy was performed. PSA values correlated significantly with the sonographically determined prostatic volumes and less precisely with the weight of the resected tissue. The PSA concentration per milliliter of prostatic volume was 0.12 ng/ml, per gram of resected tissue it was 0.21 ng/ml. An incidental PCA was found in 12/108 patients (11%). The PSA values were identical with those of the total collective in regard to volume and tissue weight. In 11 patients, we examined possible alterations of the PSA values before and until 24 h after prostatic massage. Only insignificant alterations were seen, a massive increase was not found in any patient. Searching for an absolutely valid 'normal value' appears hardly appropriate. However, the usefulness of PSA is increased when the sonographically determined prostatic volume is included. A rectal examination of the prostate has no influence on the PSA value.


Prostate-specific antigen and prostate acid phosphatase in the detection of early prostate cancer and the prediction of regional lymph node metastases. Serum values of prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) were determined in 180 patients prior to pelvic lymphadenectomy and radical prostatectomy and in 40 patients prior to pelvic lymphadenectomy alone. In all tumor stages, PSA was superior to PAP in detecting cancer of the prostate. By PSA determination using a cutoff level of 4 ng/ml (Tandem assay), 28.8% of the patients with prostate cancer, stage pT2pN0M0, and 17.8% of the cases with a stage pT3pN0M0 tumor could not be detected. All these tumors had been noticed, however, by digital rectal examination. This indicates that PSA determination cannot replace digital rectal examination as a screening method for prostate cancer. In this study, it was possible neither by PSA nor by PAP to define a practicable cutoff level for patients with and without lymph node metastases. A clear differentiation between the stages pT2pN0M0 and pT3pN0M0 was not possible by either PSA or PAP alone.


Prediction of pelvic lymph node metastases by a prostate-specific antigen and prostatic acid phosphatase in clinical T3/T4M0 prostatic cancer. Prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) were determined in the serum of 69 patients with clinical T3/T4M0 prostatic cancer before staging lymphadenectomy. In principle, high-dose radiotherapy was given only to patients of pathological N0 category. Seventeen patients had a prelymphadenectomy PSA level below the normal upper reference limit (10 micrograms/l) and only 3 of them had pelvic lymph node metastases. Fifteen of 52 patients with a preoperative PSA level > or = 10 micrograms/l were of N0 category. Only 8 of the 41 evaluable patients had PAP values above the normal range, and 6 of these 8 patients had pelvic lymph node metastases. Preoperative PSA values, but not preoperative PAP levels, assist the clinician in predicting regional lymph node metastases in patients with clinical T3/T4M0 prostatic cancer. Two-thirds of the patients with T3/T4 tumours and PSA values between 10 and 50 micrograms/l have regional lymph node metastases. About 80% of the patients with PSA levels < 10 micrograms/l belong to the N0 category. About 75% of the patients with PSA > 50 micrograms/l have N+ disease. Taking into account the individual preoperative PSA values, the indication for preradiotherapy staging lymphadenectomy should be balanced between the chance of demonstrating N+ disease, the expected postoperative morbidity and the benefit for the patient found to be of N0 category.


Transurethral radiofrequency heating or thermotherapy for benign prostatic hypertrophy: a prospective trial on 65 consecutive cases. 65 consecutive cases with symptomatic benign prostate hypertrophy were treated with transurethral radiowave thermotherapy (TURF) using the Thermex-II at a temperature of 44.5 degrees C. We report uroflowmetry and symptom scores after a follow-up of 6 months. The mean age was 63 years, the mean maximal flow (Qmax) was 8.3 ml/s, mean voided volume was 195.6 ml, mean residual urine was 60.7 ml and the mean Madsen symptom score 9.7. Values at baseline and at 1, 3 and 6 months after TURF are discussed. The mean subjective symptom score dropped to 6.3 (p < 0.01). In the entire group, the objective measure scores (OMS) did not change significantly. However the OMS improved significantly in a subgroup with a Qmax < 10.0 ml/s. The Madsen symptom score improved in 70% of the cases after 6 months.


The 'female prostate': location, morphology, immunohistochemical characteristics and significance. Using immunostaining for prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP), we can demonstrate prostate-analogous periurethral glands (the 'female prostate') in 66.7% of 33 cases. Histomorphologically, these glands resemble strongly the male prostate glands before puberty. They remain immature throughout life from the fetal period up to advanced age obviously because of a lack of an androgenic stimulus. The glands have long ducts leading into the urethra. A proper stroma component is missing. The immunohistochemical behavior of these glands also corresponds widely to that of the male prostate glands before puberty. No indications can be found for a proper biological function.