[Approaches to prevention and treatment of arterial hypoxemia in the pre-perfusion period of radical correction of Fallot's tetrad]. Examination of central hemodynamics in patients in the preperfusion period of radical correction of Fallot's tetrad has demonstrated that after pericardiotomy and the onset of surgical manipulation on the heart there was a depression of circulation both during the use of intravenous total anesthesia on the basis of ketamine and ataralgesia. The depression was more manifest under ataralgesia. The depression of circulation was paralleled by the deepening of initial hypoxemia. Analysis of the efficacy of medicamentous agents with regard to the specific features of intracardiac anatomy of the outlet of the right ventricle, estimated intraoperatively, has established that administration of phenylephrine at a rate of 3 micrograms/kg favoured an increase of partial oxygen pressure and arterial blood saturation with hemoglobin whatever the anatomo-morphological type of stenosis of the outlet of the right ventricle. Propranolol promoted the improvement of arterial blood oxygenation but in the muscular type stenosis of the outlet of the right ventricle. Meanwhile the use of the drugs possessing a beta-adreno-stimulating effect brought about the deepening of hypoxemia.


[Role of surfactant in the etiology of thrombohemorrhagic syndrome: problems of electrocoagulology]. The authors propose three hypotheses concerning heparin containing proteoglycan hypophase of surfactant. The source of thrombohemorrhagic syndrome (THS) they see in fall of the negative charge of the cells and tissues. The treatment of THS was performed by means of negative aero-ionization which turned effective in children after abdominal surgery, in burn and critical care units.


[Role of specific brain peptide factors in the pathogenesis and compensation of neurologic disorders and their use in the treatment of nervous system diseases]. New concepts of the molecular mechanisms implicated in the pathogenesis and compensation of central motor disorders are presented. It has been shown experimentally that specific peptide, postural asymmetry, and inactivation factors, play the leading part in these processes. The detection of these specific factors in the cerebrospinal fluid (CSF) of patients and convalescents allowed one to work out and test clinically a new method of treatment (CSF therapy) of patients with central motor disorders of varying genesis. The results of the clinical observations of two patients' groups (with cerebrovascular brain lesions and multiple sclerosis) are provided.


[Intracranial distension in neurosurgical pathology]. In contrast to the classic research into the problem of intracranial pressure based on the assumption that pressure in the cranial cavity is distributed uniformly, the initial methodological principle of the present work rests on the concept of the nonuniform distribution of pressure in the intracranial system both in health and disease. Comparative dynamic measurements of cerebrospinal fluid and brain tissue pressures were carried out in the early postoperative period to reveal absolute magnitudes of pressures and their correlations. Under examination were 166 neurosurgical patients with different levels of brain injury. The magnitude of cerebrospinal fluid pressure was demonstrated to depend on the level of brain injury. In the groups examined, intracerebral pressure of interstitial fluid in "normal" brain tissue was approximately the same (from -3 to +2 mm Hg). In brain edema, that pressure increased, sometimes to a considerable measure (up to 50 mm Hg). Neurosurgical pathology was shown to be characterized by nonuniform distribution of pressure in the main intracranial media: brain tissue and cerebrospinal fluid, which disturbs the physiological ratio of these pressures. It should be mentioned that some of the structures are under normal pressure whereas the other ones under elevated or lowered as compared to the physiological norm. Such a state can be correctly characterized by the term "intracranial distension" bearing in mind nonuniformity of tension in the intracranial system, a possible one-staged coexistence in the crane of the areas with elevated, normal and lowered pressures.


[A comparative experimental study of pathogenic properties of new strains of California encephalitis virus serogroups]. Experiments in noninbred mice, Syrian hamsters and grey monkeys were made to characterize the pathogenic properties of new strain of the California encephalitis serogroup isolated for the first time on the island of Taimir, in the Murmansk, Leningrad, Tver regions and in Karelia. All the strains displayed marked tropism for the CNS. The strains isolated in the northern regions of this country turned out more pathogenic for the animals. The strain Leiv-12812 Kl (isolated in the Tver region) differed from the remaining ones in more pronounced pathogenicity for the monkeys, as well as in the site and intensity of brain lesions. The persistence of the virus in hamsters organs may play a role of a reservoir of infection in nature.


[Role of enkephalins in the mechanism of anti-arrhythmia effects of adaptation in acute myocardial ischemia]. Adaptation of rats to cold and physical exercise prevented ventricular fibrillation (VF) caused by the occlusion of the left anterior coronary artery. In the heart of adapted rats with acute myocardial ischemia, myocardial enkephalins increased whereas the level of cAMP declined as compared to nonadapted animals. Injection of dalargin before the occlusion of the coronary artery in rats prevented both VF and a decrease of VF threshold. The peptide averted the rise of cAMP content in the heart during acute myocardial ischemia. The data obtained suggest that the rise of endogenous myocardial enkephalins may have an important role in antiarrhythmic action of adaptation. It is assumed that antiarrhythmic effect of enkephalins may be related to the restriction of sympathetic influence on the heart.


[Status metric analysis of epidemiological data on congenital developmental defects caused by environmental pollution]. Status metry was used to determine the portion of congenital developmental abnormalities (CDA) of obscure etiology (probably of mutation origin) among the total number of CDA in 3 cities of the Ukraine. The contribution of CDA determined by new mutations correlates with the reported data and corresponds with the level of atmospheric air pollution in the cities under study. Status metry made it also possible to define the tendencies and magnitude of the influence of factors involved in CDA formation. The conclusion is made about adequacy of the use of status metry for indirect estimation of the rate of the mutation process and its modification under the effect of environmental pollution.


[Possibility of selective binding of HIV-1 infected cells by immunosorbents]. A study was made of a possibility of specific sorption of HIV-1 infected cells with gamma-fraction of AIDS patients' serum containing high titer-specific antibodies, immobilized on the silica matrix (C-3). The laboratory tests were made with the use of the monocyte culture [symbol: see text] chronically infected with HIV-1. In addition to a decrease of the cell count after sorption, there was a decline of the activity of antigen material in the samples (to 43% of the initial). The selectivity rates were 0.92 for immunosorbents and 0.29 for sorbents of the control group, that confirms and increase of the delay of the cells expressing HIV-1 proteins on a column with the immobilized gamma-fraction of AIDS patients' serum. The use of the synthesized immunosorbents in vivo may appear debatable. However, this method can be employed for preparative purposes.


[Dietary intake of Cs-137 and Sr-90 by the population of the Gomel region, Byelarus, 1986-1989]. The authors provide the results of measuring the content of radionuclides in foods in the first 4 years after the Chernobyl accident and the data on the diet of the population. Pollution of foods was demonstrated to decrease with time. The supply of 137Cs to the body fell 3-fold from 1986 to 1989. The rejection of foods in the most polluted Vetkovsk and Narovlyansk regions made it possible to reduce 3-6-fold the mean daily supply of 137Cs with the diet and up to 2-fold the supply of 90Sr.


[Glucocorticoid receptor mechanism of regulation of the activity of angiotensin-converting enzyme and new prospects in the treatment of cardiovascular diseases]. Based on the concept of glucocorticoid-receptor induction of angiotensin-converting enzyme (ACE), approaches to inhibiting enzyme induction with drugs that suppress the function of type II cytoplasmic glucocorticoid receptors, (genuine glucocorticoid receptors), are suggested. Three types of inhibiting the function of type II glucocorticoid receptors by drugs were distinguished. Type I is characterized by competition of the drugs with natural and synthetic glucocorticoids for interaction with glucocorticoid receptors (cortexolone, progesterone); type II is determined by irreversible inactivation of type II glucocorticoid receptors (aminazine, tisercin); type III is related to an increase of interaction of transcorticoid receptors with natural glucocorticoids which is accompanied by a reduction of the interaction of natural glucocorticoids with genuine glucocorticoid receptors (analgin, amidopyrine). It has been established that the drugs that provoke irreversible inactivation of the function of type II glucocorticoid receptors decrease ACE activity in blood plasma and in the lungs, that may serve the main reason for their high hypotensive effect in arterial hypertension. A concept is advanced, providing evidence for the use of the classical ACE inhibitors and of type II glucocorticoid receptor inhibitors.


Antenatal maternal serum screening for Down's syndrome: results of a demonstration project. To assess the implementation of antenatal screening for Down's syndrome in practice, using individual risk estimates based on maternal age and the three serum markers: alpha fetoprotein, unconjugated oestriol, and human chorionic gonadotrophin. Demonstration project of Down's syndrome screening; women with a risk estimate at term of 1 in 250 or greater were classified as "screen positive" and offered diagnostic amniocentesis. Hospital and community antenatal clinics in four health districts in London. 12,603 women of all ages with singleton pregnancies seen between February 1989 and the end of May 1991, with follow up of the outcome of pregnancy completed to the end of 1991. Uptake of screening, detection rate for Down's syndrome, false positive rate, odds of being affected given a positive result, and uptake of amniocentesis in women with positive screening results, together with the costs of the screening programme. The uptake of screening was 74%. The detection rate was 48% (12/25), and the false positive rate was 4.1%, consistent with results expected from previous work based on observational studies. There was a loss of detection due to the selective use of ultrasound scans among women with positive screening results. One affected pregnancy occurred among 205 reclassified as negative; this illustrated the danger of false negatives occurring in this group and lends weight to the view that if an ultrasound estimate of gestational age is used it should be carried out routinely on all women rather than selectively among those with positive results. The estimated cost of avoiding the birth of a baby with Down's syndrome was about 38,000 pounds, substantially less than the lifetime costs of care. Antenatal maternal serum screening for Down's syndrome is effective in practice and can be readily integrated into routine antenatal care. It is cost effective and performs better than selection for amniocentesis on the basis of maternal age alone.


Opposing effects of heparin with TGF-beta or aFGF during repair of a mechanical wound of human endothelium. Influence of cAMP on cell migration. The effects on vascular wound repair in vitro of aFGF and TGF-beta, growth factors having opposite influences on endothelial cell growth and angiogenesis, were studied using as a model a mechanical lesion of confluent endothelium. Modulation by heparin of the activities of these growth factors during the repair process was also examined. Whereas heparin alone inhibited repair by lowering both cell proliferation and cell migration, TGF-beta alone mainly inhibited cell proliferation. When added together, TGF-beta and heparin exerted a combined inhibitory effect resulting in a residual lesion 50% larger than in controls. aFGF alone accelerated lesion coverage and this effect was enhanced by 40% over control values when heparin was added with aFGF. This acceleration was slightly (less than 10%) but consistently diminished by TGF-beta. Cell density in confluent unwounded areas was increased by 40% in the presence of aFGF, but TGF-beta diminished cell density by 20%. A small (30%) increase in intracellular cAMP was measured whenever aFGF was present during the repair process. In comparison, intracellular cAMP inducing agents (forskolin, dbcAMP) accelerated cell migration by 20% during lesion recovery without affecting cell proliferation or density. The present results show that the inhibitory effects of TGF-beta during vascular wound repair are opposed by aFGF. Furthermore, heparin (or heparan sulfates in vivo) modulates growth factors having activating or inhibiting functions and thus plays a regulatory role during the repair process. cAMP-inducing substances other than growth factors are able to accelerate cell migration.


Child mental health in the Americas: a public health approach. The systematic, population-wide application of preventive measures based on what is known about the causes and outcomes of psychiatric disorders can markedly reduce morbidity from mental ill health among children in the Americas. The actions proposed here rely partly upon increasing access for all women and their children to thoroughly tested obstetric and pediatric care; in part they depend on improving nutrition and opportunities for cognitive stimulation; and in part they call for enhancing the mental health skills of primary care practitioners by appropriate in-service training. There are limits to our knowledge and to the effectiveness of some of our interventions; nonetheless, the greatest barrier to better child mental health is failure to muster the political will to apply what is known to the care of mothers and children in all sectors of society.


Involvement of substance P present in primary afferent neurones in modulation of cutaneous blood flow in the instep of rat hind paw. 1. The participation of small-diameter afferent fibres in the microcirculatory haemodynamics of cutaneous tissue was examined by studies on the effects of antidromic stimulation of primary afferent neurones on cutaneous blood flow (CBF) and tachykinin release into the subcutaneous space in the instep of the hind paw of rats. 2. Antidromic stimulation of the sectioned sciatic nerve induced a biphasic flow response, an initial transient decrease followed by an increase, with no alteration in the blood pressure. 3. Neither phase was affected by pretreatment with phentolamine (0.1 mg kg-1, i.a.), propranolol (0.5 mg kg-1, i.a.), atropine (0.5 mg kg-1, i.a.), methysergide (0.5 mg kg-1, i.a.) or mepyramine (10 mg kg-1, i.a.) plus cimetidine (10 mg kg-1, i.a.), but both were significantly inhibited by pretreatment with capsaicin (50 mg kg-1, s.c.). 4. Spantide (1-2 mumol kg-1, i.a.), a substance P (SP) antagonist, reduced the basal CBF, and also inhibited both phases of the biphasic flow response evoked by antidromic stimulation of the sectioned sciatic nerve. 5. Intra-arterial infusion of SP (0.5 mumol kg-1, i.a.) induced a biphasic flow response similar to that elicited by antidromic stimulation of the sectioned sciatic nerve. 6. Antidromic stimulation of the sectioned sciatic nerve caused a marked increase in SP release into the subcutaneous perfusate of the instep of the rat hind paw, but no detectable increase in neurokinin A release.7. We suggest that SP and its receptors are mainly responsible for the vascular response induced by stimulation of the sectioned sciatic nerve, and that small-diameter afferent fibres containing SP tonically regulate vascular tone in cutaneous microvessels.


Structure-activity relations of amiloride and its analogues in blocking the mechanosensitive channel in Xenopus oocytes. 1. Patch clamp recording techniques have been used to compare the block caused by amiloride and some of its structural analogues of the mechanosensitive (MS) cation selective channel in frog (Xenopus laevis) oocytes. 2. Like amiloride, the amiloride analogues dimethylamiloride (DMA), benzamil and bromohexamethyleneamiloride (BrHMA) block the MS channel in a highly voltage-dependent manner. 3. All analogues tested were more potent blockers than amiloride with IC50's of 500 microM (amiloride), 370 microM (DMA), 95 microM (benzamil) and 34 microM (BrHMA). 4. Hill plots gave Hill coefficients of 2 (amiloride), 1.8 (DMA), 1 (benzamil) and 1.2 (BrHMA) indicating that the binding of two ligand molecules may be necessary for the block caused by amiloride, DMA and possibly BrHMA whereas only a single ligand molecule may be required for the block by benzamil. 5. The potential use of BrHMA as a light-activated, covalent label of the MS channel protein is discussed. 6. The amiloride analogue 'fingerprinting' of the blocking site on the MS channel indicates it is structurally different from previously described amiloride-sensitive ion transport pathways but may be related to the amiloride binding site on outer hair cells of the ear.


Evidence for a glutamate receptor of the AMPA subtype which mediates insulin release from rat perfused pancreas. 1. The effect of L-glutamate has been studied on insulin secretion by the isolated perfused pancreas of the rat. The glutamate receptor subtype involved has been characterized. 2. In the presence of a slightly stimulating glucose concentration (8.3 mM), L-glutamate (5 x 10(-5)-4 x 10(-3) M) induced an immediate, transient and concentration-dependent insulin response. On the other hand, in the presence of a non stimulating glucose concentration (2.8 mM), L-glutamate (10(-3) M) did not modify the basal insulin secretion. 3. The three non-NMDA receptor agonists, kainate (10(-4)-10(-3) M), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA, 5 x 10(-5)-10(-4) M) and quisqualate (5 x 10(-6)-5 x 10(-5) M) all provoked a transient and concentration-dependent insulin response from pancreas perfused with 8.3 mM glucose. Compared with glutamate, kainate exhibited a similar efficacy, whereas AMPA and quisqualate elicited only a 3 fold lower maximal insulin response. In contrast, NMDA (10(-4)-10(-3) M) was ineffective. 4. An antagonist of non-NMDA receptors, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 5 x 10(-5) M) totally prevented the stimulatory effect of L-glutamate (4 x 10(-4) M) and kainate (2 x 10(-4) M). In contrast, the NMDA receptor antagonist, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine ((+) MK801) was without effect. 5. The insulin secretory effect of glutamate (4 x 10(-4) M) was not affected by atropine (3 x 10(-7) M) or tetrodotoxin (3 x 10(-6) M). 6. Quisqualate at a high maximally effective concentration (4 x 10(-4) M) inhibited glutamate (10(-3) M) or kainate (4 x 10(-4) M)-induced insulin release. 7. This study shows that L-glutamate stimulates insulin secretion in rat pancreas, by acting on an excitatory amino acid receptor of the AMPA subtype.


Role of the hospital support team.

Role of the hospital support team. The hospital support team is a multidisciplinary team through which the expertise of hospice care can be extended to the acute hospital situation. This article highlights the aims of such a team and the expertise of different members, emphasizing the advisory role and the importance of 'role-modelling'.


Cholinoceptor regulation of cyclic AMP levels in bovine adrenal medullary cells. 1. The regulation of adenosine 3':5'-cyclic monophosphate (cyclic AMP) levels by cholinoceptors has been studied in cultured bovine adrenal medullary cells. 2. Acetylcholine (100 microM), nicotine (10 microM) and dimethylphenylpiperazinium (20 microM) each increased cellular cyclic AMP levels 2 to 4 fold over 5 min in the absence of phosphodiesterase inhibitors. The muscarinic agonist acetyl-beta-methylcholine (100 microM) had no effect either on its own or on the response to nicotine. The responses to acetylcholine and nicotine were unaffected by atropine (1 microM) but were abolished by mecamylamine (5 microM). 3. Cellular cyclic AMP increased transiently during continuous exposure to nicotine (1-20 microM), with the largest response seen after 5 min, a smaller response after 20 min, and no change in cyclic AMP levels seen after 90 or 180 min. The maximal response after 5 min stimulation was seen with 5-10 microM nicotine and the EC50 was about 2 microM. In contrast, extracellular cyclic AMP levels did not change after 5 or 20 min stimulation with nicotine, but increased slightly after 90 min and further after 180 min. 4. The cellular cyclic AMP response to nicotine (10 microM) was unchanged or weakly enhanced in the presence of the unselective phosphodiesterase inhibitor, isobutylmethylxanthine, and was unchanged in the presence of rolipram. Nicotine did not interact synergistically with low concentrations of forskolin. The response was however completely abolished in the absence of extracellular Ca2+.


Pharmacological characterization of non-NMDA subtypes of glutamate receptor in the neonatal rat hemisected spinal cord in vitro. 1. A grease-gap technique was used to record depolarizing responses to alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionate (AMPA), kainate and N-methyl-D-aspartate (NMDA) in the hemisected spinal cord of the neonatal rat. The pharmacology of non-NMDA subtypes of glutamate receptor was investigated with the novel quinoxalinedione, 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo (F)-quinoxaline (NBQX) and with a series of barbiturates. 2. NBQX antagonized AMPA- and kainate-, but not NMDA- induced depolarizations. The near parallel shifts of the major part of the dose-response curves for AMPA and kainate by NBQX gave pA2 values (+/- s.e.) of 6.7 +/- 0.2 and 6.8 +/- 0.2 respectively, consistent with a common site of action for these two agonists. 3. Below the 50% level at which these pA2 values were calculated, however, an NBQX-resistant plateau was seen within the kainate, but not the AMPA, dose-response curve. 4. In decreasing order of potency, methohexitone, secobarbitone, thiopentone, pentobarbitone and phenobarbitone preferentially reduced kainate-, rather than AMPA- and NMDA-, induced depolarizations. Methohexitone was also the most selective with IC50S against kainate, AMPA and NMDA of 31 +/- 7, 172 +/- 47 and greater than 200 microM respectively. 5. The NBQX-resistant plateau seen within the kainate dose-response curve was reduced by methohexitone. Kainate antagonism by methohexitone was not reduced by 50 microM picrotoxin. 6. We conclude that, while mixed agonist actions may hamper demonstration of antagonist selectivity, depolarizations induced by the non-NMDA ionotropic agonists, AMPA and kainate, are mediated in part via distinct receptors.


Effects of a potassium channel opener (SDZ PCO 400) on guinea-pig and human pulmonary airways. 1. SDZ PCO 400 evoked dose-related relaxation of isolated airway smooth muscle. For human bronchus precontracted by endogenous tone or addition of carbachol (10(-5) M), IC50 values were 1.74 microM and 1.82 microM respectively. With guinea-pig trachea contracted by endogenous tone, a comparable IC50 (1.79 microM) was observed, but no IC50 (less than 100 microM) could be determined following contraction by carbachol (10(-6) M). 2. Airway obstruction induced by intravenous bombesin in the anaesthetized ventilated guinea-pig was diminished by intravenous injection of SDZ PCO 400 (ID50 54 micrograms kg-1) or by introduction into the duodenum (ID50 1.0 mg kg-1). Inhalation of nebulized SDZ PCO 400 (0.1 mg kg-1) diminished airway obstruction due to intravenous injection of histamine (3.2-5.6 micrograms kg-1) for up to 20 min. 3. Increased bronchoconstrictor responses to bombesin (180-240 ng kg-1) following intravenous infusion of platelet activating factor (PAF) or (+/-)-isoprenaline, or to histamine (1.0-3.2 micrograms kg-1) following intravenous injections of immune complexes, were suppressed following concomitant intravenous infusion of SDZ PCO 400 (ID50 0.3 mg kg-1 h-1, 1.0 mg kg-1 h-1 and 0.1 mg kg-1 h-1 respectively). 4. Intravenous injection of SDZ PCO 400 (0.1 mg kg-1) effected transient (less than 10 min) inhibition of histamine-induced bronchospasm, yet diminished, for prolonged periods [up to 40 min] the enhanced bronchoconstrictor responses to histamine that followed intravenous injections of immune complexes.The capacity of SDZ PCO 400 to resolve such established airway hyperreactivity was prevented by prior intraduodenal instillation of a potassium channel antagonist, glibenclamide (30 mg kg-').5. In sensitized guinea-pigs, SDZ PCO 400 inhaled as a dry powder (5.7 mg kg-') suppressed development of allergic airway hyperreactivity to histamine (1.8-3.2;pg kg-', i.v.), but failed to diminish accumulation of eosinophils or other inflammatory cells within the airway lumen 24 h after inhalation of ovalbumin.6. Preincubation (30 min) of isolated sensitized trachea of guinea-pig with SDZ PCO 400 (10-5-10-4M) did not influence contractile responses to ovalbumin. However in anaesthetized sensitized guinea-pigs,insufflation of SDZ PCO 400 (1.25 mg) as a powder substantially diminished airway obstruction that followed inhalation of ovalbumin. This effect was prevented by prior vagal section.7. It is concluded that SDZ PCO 400 reduces airway obstruction not only through direct actions on airway smooth muscle but also by impairing the expression of airway hyperreactivity, without directly influencing inflammatory events in the airways.


Effects of imipramine on the transient outward current in rabbit atrial single cells. 1. The effects of imipramine on action potential characteristics and transient outward potassium current (It) of rabbit isolated atrial myocytes were studied using the whole-cell configuration of the patch-clamp technique. 2. Imipramine, 3 microM, decreased action potential amplitude and lengthened the action potential duration measured at 50% of repolarization, whereas it did not modify the final phase of repolarization or the resting membrane potential. These results are similar to those reported in multicellular rabbit atrial preparations. 3. Imipramine, 0.1-100 microM, induced a concentration-dependent inhibition of the peak amplitude of It, a shortening of the time to peak current and an increase in the inactivation rate. The acceleration of the current inactivation is to a major extent responsible for the decrease in the integral of the outward current measured at 50 ms after the start of the pulse. 4. The drug-induced block of It was not associated with changes in the voltage-dependence of the steady-state inactivation curve or in the process of recovery from inactivation of the current. Extrapolation to zero block shows that imipramine did not block It before its activation at the onset of the depolarization. These results suggested that imipramine does not affect the inactivated or the resting state of It channels. 5. It is concluded that in rabbit isolated atrial cells, imipramine inhibits It and that this effect is responsible for the lengthening of the action potential duration produced by this drug.


Effect of the beta-adrenoceptor agonist clenbuterol and phytohaemagglutinin on growth, protein synthesis and polyamine metabolism of tissues of the rat. 1. The kidney bean lectin, phytohaemagglutinin (PHA), induced a marked atrophy of skeletal muscle which was evident from the changes in tissue composition (protein, RNA, DNA and polyamine content) and from the reduction in weight and protein synthesis of hind leg muscles of rats fed on kidney bean-diets for four days. The beta-adrenoceptor agonist, clenbuterol, induced skeletal muscle hypertrophy by transiently stimulating protein synthesis. As a consequence, the muscle loss caused by a short exposure to PHA was, in part, ameliorated by clenbuterol treatment. 2. Cardiac muscle was affected to a lesser extent than skeletal muscle by both clenbuterol and the lectin. However, there was evidence that protein synthesis in heart was reduced by PHA. 3. PHA had opposite effects on the gut, the lectin-induced hyperplasia of the jejunum was accompanied by a large increase in protein synthesis. Clenbuterol alone had no effect on the jejunum whereas a combination of PHA and clenbuterol appeared to exacerbate the effect of the lectin on gut. 4. Both the lectin-induced gut growth and the hypertrophy of skeletal muscle caused by clenbuterol were preceded by the accumulation of polyamines in the respective tissues. Of particular note was the observation that a significant increase in the proportion of the intraperitoneally injected 14C-labelled spermidine or putrescine taken up by the growing tissues could be detected by the second day. Therefore, the measurement of uptake of labelled polyamines may be used as a sensitive indicator of early alterations in tissue metabolism.


Ca2+ release from isolated sarcoplasmic reticulum of guinea-pig psoas muscle induced by K(+)-channel blockers. A Ca(2+)-sensitive electrode was used to measure the Ca2+ concentration of the medium containing the heavy fraction of the fragmented sarcoplasmic reticulum (SR) prepared from guinea-pig psoas muscle. Among K(+)-channel blockers tested, 4-aminopyridine (4-AP), tetraethylammonium (TEA) and charybdotoxin elicited Ca2+ release from the SR, but apamin and glibenclamide did not. These results suggest that a reduction of SR K+ conductance leads to Ca2+ release from the SR.


The neuroprotective effect of a nitric oxide inhibitor in a rat model of focal cerebral ischaemia. Recent data showed that glutamate toxicity in primary cortical cultures is mediated by nitric oxide. In order to investigate the effect of inhibition of NO synthase on focal cerebral ischaemia in rats, we studied the histological consequences of a middle cerebral artery (MCA) occlusion after post-operative treatment with NG-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase. We found a significant reduction of cortical (-43%) and striatal (-25%) necrotic volumes induced by MCA occlusion, indicating that NO synthesis plays an important role in the neurotoxic cascade leading to neuronal damage after focal cerebral ischaemia in rats.


Characterization and localization of nitric oxide synthase in non-adrenergic non-cholinergic nerves from bovine retractor penis muscles. 1. Partially purified soluble nitric oxide (NO) synthase was isolated from the bovine retractor penis muscle (BRP), a tissue in which the inhibitory response to non-adrenergic non-cholinergic nerve (NANC) stimulation appears to be mediated by NO or NO-like material. 2. NO synthase from BRP used L-arginine as a substrate, required NADPH, tetrahydrobiopterin, and FAD as co-factors and was Ca2+/calmodulin-dependent. The activity of NO synthase was inhibited by NG-methyl-L-arginine and NG-nitro-L-arginine, and haemoglobin blocked the effect of NO formed by the enzyme. 3. On reducing SDS polyacrylamide gel electrophoresis the apparent molecular mass of NO synthase from BRP was 160 +/- 2 kDa, which is similar to that of the cerebellar NO synthase. Protein immunoblot and immunoprecipitation showed that NO synthase from BRP cross-reacted with the selective antiserum to neuronal NO synthase from rat cerebellum. 4. Immunohistochemistry using the same antiserum demonstrated that NO synthase in BRP was located exclusively within nerve fibres. Thus, autonomic nerves synthesizing the NANC neurotransmitter seem to contain an isoform of NO synthase which is similar to that from rat cerebellum.


Involvement of inflammatory mediators in the airway responses to trimellitic anhydride in sensitized guinea-pigs. 1. We examined the effect of various pharmacological agents on the acute bronchoconstrictor response and airway microvascular leakage in a model of guinea-pig sensitization to trimellitic anhydride (TMA) a cause of low molecular weight occupational asthma in man. 2. Guinea-pigs were given intradermal injections of 0.1 ml of 0.3% TMA in corn oil; 21-28 days later, anaesthetized guinea-pigs were challenged with TMA conjugated to guinea-pig albumin by tracheal instillation. Changes in lung resistance were measured and airway microvascular leakage was quantified by measuring the extravasation of Evans blue dye into the airway tissue. 3. Sensitized guinea-pig (n = 9 in each group) were pretreated with chlorpheniramine (2.5 mg kg-1, i.v.), WEB 2086 (10 micrograms kg-1, i.v.), BW 4AC (50 mg kg-1, i.p.), nedocromil sodium (2% aerosol for 60 s) or vehicle alone. 4. Pretreatment with chlorpheniramine inhibited both the acute bronchoconstrictor response and the increase in airway microvascular leakage. WEB 2086 and nedocromil sodium partially inhibited the bronchoconstrictor response but had no significant effect on airway microvascular leakage. BW 4AC caused a non-significant reduction of the bronchoconstrictor response and airway microvascular leakage. 5. These results indicate that both the bronchoconstrictor response and the airway microvascular response in this model of sensitization is mediated to a large extent by histamine. PAF but not 5-lipoxygenase products also partially mediates the bronchoconstrictor response but not the airway microvascular leakage. Nedocromil sodium partially inhibits the bronchoconstrictor response only.


The cardiodepressant and vasodepressant effects of tumour necrosis factor in rat isolated atrial and aortic tissues. 1. The ability of recombinant human tumour necrosis factor-alpha (rec huTNF) to elicit cardiodepressor and vasodepressor effects in rat isolated tissues was investigated. 2. rec huTNF (3 x 10(-11)-3 x 10(-8) M) administered directly to the organ bath, caused a concentration-dependent relaxation of the isoprenaline-induced inotropic response in electrically stimulated rat left atria. This occurred within 20 min of administration. In contrast, rec huTNF was without effect on the chronotropic response to isoprenaline in isolated spontaneously beating atria. 3. rec huTNF (1 microgram kg-1) was also given systemically to rats and the atria studied in vitro. Only 60 min of rec huTNF pretreatment was sufficient to cause a marked attenuation of the isoprenaline-induced inotropic response. This effect was not further augmented when rats were pretreated with rec huTNF for 24 h. 4. In isolated aortic rings taken from rats 60 min after rec huTNF (1 microgram kg-1, i.v.) administration, there was no effect seen on the constriction induced by phenylephrine in either endothelium-intact or denuded tissues. In addition, any responses to L-arginine or NG-nitro-L-arginine methyl ester (L-NAME) administration were unaffected by rec huTNF pretreatment. 5. In aortic rings taken from rats 24 h after rec huTNF administration, the phenylephrine-induced constriction was significantly attenuated in tissues with an intact endothelium. Furthermore, the relaxation to subsequent L-arginine administration was greater in these tissues than in those saline-treated rats. In addition, in both endothelium-intact and denuded tissues, the vasoconstrictor response to L-NAME (10-3M) was significantly augmented. 6. These data suggest that rec huTNF possesses both cardiodepressant properties with a rapid onset of action and vasodepressant properties with a slow onset of action. The latter could be mediated through the induction of a non-constitutive form of the NO-synthase enzyme present within the vascular wall.


Endotoxin-induced vasodilatation in anaesthetized rat skin involves nitric oxide and prostaglandin synthesis. 1. The effect of intradermally injected endotoxin on skin blood flow was investigated in anaesthetized male Wistar rats in vivo. 2. Local skin blood flow changes were measured hourly for 6 h in the shaved dorsal skin with a laser-Doppler flow probe and compared to changes in control sites which had been injected with 100 microliters of phosphate-buffered saline. By 3 h, skin blood flow increased above basal by 129 +/- 27% and 186 +/- 29% with 1 and 10 micrograms of endotoxin respectively. Blood flow remained significantly elevated at 6 h, the corresponding figures being 129 +/- 24% and 154 +/- 31% (P less than 0.05, n = 6 rats, mean +/- s.e.mean). 3. In further experiments, the response to 3 micrograms of endotoxin was measured at 4 h and treatment with a cyclo-oxygenase inhibitor, nitric oxide synthase inhibitors or a topical steroid all significantly inhibited this response (P less than 0.05 in each case, n = 6 rats in each group with duplicate sites in each animal). 4. Indomethacin 3 x 10(-9) mol per site injected 3.5 h after injection of endotoxin suppressed the mean 4 h response to endotoxin by 78%; NG-nitro-L-arginine methyl ester (L-NAME) 10(-7) mol per site suppressed the response by 95%; NG-monomethyl-L-arginine (L-NMMA) 10(-7) mol per site suppressed the response by 50%; whereas the D-isomer of NG-monomethyl-arginine 10(-7) mol per site had no significant effect.5. Topical application of the corticosteroid, betamethasone 17-valerate (1% solution) 18 h before injection of endotoxin inhibited the mean 4 h response to endotoxin by 66% and the 6 h response by 48%.6. In the same model, the vasodilator response to arachidonic acid was inhibited by both indomethacin and nitric oxide synthase inhibitors (P<0.05 in each case).7. These data suggest that the microcirculatory vasodilator response to endotoxin and arachidonic acid injected locally involves both nitric oxide synthase and cyclo-oxygenase in this in vivo model.


Urological audit: the role for an aggressive approach to high grade superficial bladder tumours. A retrospective study was undertaken of the different treatment modalities for bladder tumours under the care of 3 consultants in the urology department of a district general hospital. The aim was to review the results of the various forms of treatment. In all, 261 patients' case records were reviewed and 19 variables extracted. There was an average delay of 4.2 months from the onset of symptoms to the initial cystoscopy. Over 50% of high grade tumours were invasive on initial presentation (G3T2/G3T3). A range of treatments for the more aggressive tumours was adopted by the urologists, ranging from a conservative resection (TURBT) to an aggressive approach (cystourethrectomy) at the earliest sign of progression. A strong association between aggressive treatment and higher survival was noted. This study has proved valuable in demonstrating to the urology team the value of routine audit and questioning "established" surgical practice. As a result, a more standard regime for the treatment of bladder tumours has been advocated and a prospective randomised controlled trial will be introduced.


Transurethral incision versus transurethral resection of the prostate. A subjective and objective analysis. A prospective study was undertaken comparing transurethral incision of the prostate (TUIP) with transurethral resection (TURP) in the treatment of 220 patients with urinary obstruction caused by a small, benign prostate. Patients were managed alternately by TUIP and TURP, and their symptoms and urodynamic findings evaluated before and after surgery. Subjectively and objectively, the results were comparable in both groups. Pre- and post-operative complications were significantly less for the TUIPs than the TURPs. TUIP was significantly better than TURP in terms of shorter operating time, duration of hospitalisation and reduced need for transfusion. We recommend TUIP as the operation of choice for the relief of obstruction in the presence of a small, benign prostatic enlargement.


Localised deep microwave hyperthermia in the treatment of benign prostatic hyperplasia: long-term assessment. A group of 133 patients with benign prostatic hyperplasia who were either poor operative risks or who had refused surgery underwent localised deep microwave hyperthermia, without supplementary drugs. In 59% of patients who had had an indwelling catheter, freedom from urological obstruction and satisfactory voiding were maintained for 7 years and catheterisation was not required. In patients with severe prostatic symptoms, 65% showed general improvement and satisfactory voiding for 8 years. There were no side effects. Patients who relapsed were given a second course of treatment and 75% responded positively. This positive response and the lack of side effects suggest that localised deep microwave hyperthermia may be an effective alternative to surgery in the management of high risk patients and those who are reluctant to undergo surgery.


Inter-relation between measurement of serum prostatic specific antigen and transrectal ultrasound in the diagnosis of benign prostatic hyperplasia and prostatic cancer. Serum prostatic specific antigen (PSA) and ultrasound-determined prostatic volume (UPV) were measured in 50 patients with histologically proven benign prostatic hyperplasia (BPH) and in 40 patients with histologically proven prostatic cancer of whom 17 had evidence of distant metastases (M1) and 23 did not (M0). A good correlation between log PSA and UPV was demonstrated in the BPH group and rearrangement of the linear regression equation allowed calculation of a single variable--the log PSA corrected to a standard prostate volume for any given individual. A volume-corrected PSA correctly identified all patients with M1 disease and greatly improved but did not eliminate overlap of M0 disease with BPH. Reduction of serum PSA to a single volume-corrected variable will allow the introduction of practical and optimum protocols for the management of patients with prostatic enlargement.


Limitations of laser treatment for malignant dysphagia. Of 86 patients with inoperable malignant dysphagia, 68 (79 per cent) underwent successful palliation by endoscopic laser therapy, of whom 24 remained well palliated until the time of death. In 18 patients laser treatment was unsuccessful and nine of these underwent intubation, eight successfully. After successful laser therapy, dysphagia recurred in 44 patients a mean of 7.8 weeks later. Of these, 31 received palliation until death by dilatation with or without laser therapy, and 13 required intubation. The overall laser-related complication rate was 12 per cent with a mortality rate of 4 per cent. The intubation-related mortality rate was 9 per cent.


Direct surface cooling of the exocrine pancreas in the rat. A study was carried out to evaluate the effects of direct cooling on the exocrine pancreas. Changes in amylase and cathepsin B release, and in the subcellular distribution of amylase and cathepsin B were measured after 1, 2 and 3 h of direct pancreatic cooling in rats. Cooling for 2 and 3 h caused significant hyperamylasaemia and increased pancreatic amylase content, but minimal histological change. Furthermore, 3 h of cooling caused marked redistribution of cathepsin B activity from the lysosomal fraction to the heavier zymogen fraction, and co-localization of lysosomal and digestive enzymes; amylase and cathepsin B output into pancreatic juice after caerulein stimulation were also reduced. These results show that direct pancreatic cooling impairs exocrine function and implicate lysosomal enzymes in the pathogenesis of pancreatic injury, in agreement with results from other models of experimental pancreatitis.


Results of surgical palliation for cancer of the head of the pancreas and periampullary region. Between 1977 and 1986, 101 patients underwent surgical bypass for periampullary carcinoma. The hospital mortality rate was 18 per cent and the morbidity rate 43 per cent. Mortality was not influenced by the extent of the tumour. Survival rates at 1,2 and 3 years were 28, 9 and 4 per cent, respectively. The median survival time was 17 months for localized tumours, 10 months for those that had invaded surrounding tissues, 6 months in the presence of lymph node involvement and 3 months with distant metastasis. The quality of survival was good for most patients with localized tumours but poor for those with parenchymal metastasis, in whom palliation was transient for 85 per cent and effective for less than half of their survival time for 60 per cent. These results suggest that patients with distant metastasis but without impending duodenal obstruction should undergo palliation by endoscopic or percutaneous routes while those with less advanced disease or with duodenal involvement remain candidates for surgical bypass.


Induction of cobalt accumulation by excitatory amino acids within neurons of the hippocampal slice. Computer-assisted image analysis was used to establish the dose response of excitatory amino acid (EAA) analogs on the induction of cobalt accumulation within pyramidal and granule cell neurons in 400 microns slices of gerbil hippocampus. Slices were incubated 20 min at 22 degrees C in a solution containing 5 mM CoCl2 and 0-1,000 microM EAA analog. The cobalt was visualized by development in (NH4)2S, and the slices were digitized for quantitative densitometry. Kainic acid (KA) had the largest effect and induced cobalt accumulation in the dentate gyrus and CA1, 180% and 150% above control, respectively, with an ED50 = 30 microM. alpha-Amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) induced accumulations of cobalt in CA1 and hilar neurons 130% above control with an ED50 = 30 microM, but had little effect on dentate granule cells. The accumulations induced by KA and AMPA were blocked by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), but not by DL-2-amino-5-phosphonovaleric acid (AP5) or voltage-dependent calcium channel blockers. N-Methyl-D-aspartate (NMDA) induced accumulation in the dentate and CA1 150% above control in a pattern similar to KA, but with an ED50 of 100 microM. The accumulation was blocked by both AP5 and CNQX. These data indicate that cobalt-permeable, receptor-activated divalent cation channels are differentially distributed within the gerbil hippocampus, and have differential sensitivities to non-NMDA agonists. The localization of KA-activated, cobalt-permeable channels appears to be coincident with the flop form of the AMPA-selective calcium-permeable glutamate receptor-activated channel.(ABSTRACT TRUNCATED AT 250 WORDS)


Decreased neuropeptide-converting enzyme activities in cerebrospinal fluid during acute but not chronic phases of collagen induced arthritis in rats. We investigated the effects of collagen II-induced arthritis on two cerebrospinal fluid (CSF) enzymes converting dynorphin A and substance P (SP), namely dynorphin-converting enzyme (DCE) and substance P endopeptidase (SPE). The products generated by these enzymes are the bioactive fragments Leu-enkephalin-Arg6 and substance P, respectively. The strain used (DA rats) is very sensitive towards induction of arthritis. The collagen arthritis is a chronic autoimmune arthritis induced by native rat collagen type II (CII). Following intradermal injection of CII into the tailbase. CSF was sampled on day 21 (acute arthritis) and day 38 (chronic arthritis). Control rats were untreated because the strain used developed an acute and self-limited arthritis (adjuvant arthritis) when administered vehicle (i.e. incomplete Freund's adjuvant). The DCE activity was significantly lowered in the acute phase of arthritis (P less than 0.05) when analysed with two-factor analysis of variance (ANOVA). The enzyme converting SP (SPE) also showed a significant decrease in the acute phase of arthritis (P less than 0.05). These results demonstrate that both DCE and SPE are affected in the acute phase of arthritis. A functional role of these enzymes in processing pain-related neuropeptides is therefore implicated.


Melanin, tyrosine hydroxylase, calbindin and substance P in the human midbrain and substantia nigra in relation to nigrostriatal projections and differential neuronal susceptibility in Parkinson's disease. The anatomy of melanin-containing neurons and other midbrain structures was examined by tyrosine hydroxylase (TH), calbindin D28k, and substance P immunostaining. Greater than 95% of cells in the substantia nigra pars compacta contained melanin, but densely packed cells in a ventral tier had a low content of melanin and loosely packed cells in a dorsal tier had a high content of melanin. Approximately 60% in the gamma group and 40% in the retrorubral nucleus had a low content of melanin. TH immunostaining was moderate in both the ventral and dorsal tiers, but more intense in the gamma group and retrorubral nucleus. Calbindin D28k was absent from the ventral and dorsal tiers, but present in the gamma group and retrorubral nucleus. In the light of primate tracing studies these findings suggest that the ventral tier of the pars compacta projects to striosomes of the striatum and the dorsal tier, gamma group and retrorubral nucleus to the matrix compartment. The ventral tier is more vulnerable than the dorsal tier in Parkinson's disease, but the cells contain less melanin. Neither tier contains calbindin D28k. This differential vulnerability between the ventral and dorsal tiers cannot be explained by melanin or calbindin D28k.


Localization and retention in vitro of fluorescently labeled aortic baroreceptor terminals on neurons from the nucleus tractus solitarius. The anterograde fluorescent tracer DiA was used to visualize baroreceptor fibers and synaptic terminals both in living and fixed tissue. Baroreceptor fibers labeled with DiA terminated as a dense synaptic field in the medial nucleus tractus solitarius (NTS), making synaptic contact on the soma, as well as processes of neurons that they innervated. A similar distribution and morphology was observed in baroreceptor fibers and terminals labeled with horseradish peroxidase. DiA also identified baroreceptor terminals and the neurons receiving these synaptic contacts in vitro. NTS neurons were dissociated from their surrounding tissue and identified by attached baroreceptor terminals that retained the fluorescent dye. These results will enable us to study the electrophysiological properties of dispersed neurons that receive identified baroreceptor synaptic terminals.


Excitatory amino acid receptor activation produces a selective and long-lasting modulation of gene expression in hippocampal neurons. Activation of excitatory amino acid (EAA) receptors in cultured hippocampal neurons causes down-regulation of the protein ligatin, a receptor for phosphoglycoproteins and a marker protein for membrane-vesicle transport systems. This reduction occurs at both physiologic and excitotoxic levels of glutamate stimulation and is accompanied by a significant decrease in steady state levels of ligatin mRNA. Reduction in ligatin mRNA occurs within 60 min and persists 24 h later. Steady state levels of mRNAs encoding cyclophilin, an ubiquitous cytosolic protein, and neuron specific-enolase (N-SE) are not diminished by glutamate receptor activation, demonstrating that down-regulation of ligatin mRNA was not a result of general catabolism. Further, this reduction in ligatin mRNA occurred without induction of HSP 70. Pharmacological studies using selective antagonists and agonists indicate that this down-regulation of ligatin gene expression is predominantly mediated by the N-methyl-D-aspartate (NMDA) subclass of EAA receptors and that Ca2+ is required. This is the first report that EAA receptor activation in hippocampal neurons can pretranslationally down-regulate gene expression in a rapid and long-lasting manner under physiologic, as well as cytotoxic conditions. The data support the hypothesis that modulation of neuronal gene expression may represent a molecular mechanism mediating some of the long-lasting functional and pathophysiological effects of EAA on cell function.


Spontaneous and stimulated firing in cultured rat suprachiasmatic neurons. Neurons from the suprachiasmatic nucleus (SCN) of the hypothalamus, the site of a circadian pacemaker in mammals, were isolated from embryonic rat. After mechanical dissociation neurons were brought into culture for 1-2 weeks, using a chemically defined medium. Recordings were made from 74 bipolar neurons using two different configurations of the patch-clamp technique. During cell attached patch recordings, 45% of neurons fired spontaneously. The mean firing rate was 0.7 +/- 0.6 Hz and the firing pattern was irregular. In whole cell recordings 73% of the investigated neurons showed spontaneous activity with an irregular firing pattern. The mean spontaneous firing rate with an intracellular Cl- concentration of 145 mM was 1.0 +/- 0.6 Hz. The resting membrane potential of the bipolar neurons was estimated to be -62 +/- 24 mV. An intracellular Cl- concentration of 145 mM depolarised the membrane potential. It also increased the probability of spontaneous firing. A depolarising current stimulus produced an action potential with a threshold voltage of -46 +/- 9 mV. Suprathreshold stimuli resulted in repetitive firing with a mean frequency of 12 +/- 4 Hz. The minimum interspike interval was 52 +/- 14 ms. All action potentials either occurring spontaneously or elicited by current stimuli were abolished by the Na(+)-channel blocker TTX. These results indicate that our cultured neurons have some electrophysiological properties in common with SCN neurons in brain slices and in vivo.


Nerve growth factor responsiveness of cultured major pelvic ganglion neurons from the adult rat. The bladder and other pelvic viscera are innervated in the rat by the major pelvic ganglion (MPG), a mixed sympathetic/parasympathetic population of neurons that participates in lower urinary pathophysiology. Neurons from the MPG of adult females were removed, dissociated and cultured in order to test retention of the neuronal phenotype and whether they responded to Nerve Growth Factor (NGF). The bladder-specific subset of MPG neurons were distinguished by retrograde labeling prior to culture. The adult ganglionic neurons adapted to culture with greater than 80% survival in the best cases. The cultured neurons retained excitability, as determined by measuring voltage-activated ionic currents. They were positive for neuron-specific beta-tubulin and many retained immunoreactivity for characteristic peptides and transmitter synthetic enzyme. The proportion of neurons in the different categories tested varied somewhat from that in vivo, but there was no evidence of selective death of a particular population. The cultured MPG neurons were responsive to NGF and anti-NGF antibody. NGF supported neuronal survival and expression of tyrosine hydroxylase. Added NGF also affected the expression of neuropeptide Y. Hypertrophied neurons from animals with experimental bladder outlet obstruction demonstrated increased responsiveness to NGF. The data suggest that NGF participates in adult neural plasticity due to continued responsiveness to the factor. Furthermore, questions concerning regulation of MPG neurons may be addressed in vitro.


Olfactory bulb transplantation into the olfactory bulb of neonatal rats: a WGA-HRP study. After unilateral bulbectomy in neonatal (P1-P5) rats, autoradiographically prelabeled presumptive olfactory bulbs from E15 and E17 embryos were transplanted in place of the removed tissue. After 2-7 months, the animals received injections of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the piriform cortex. Nine of the twenty animals revealed WGA-HRP-positive neurons among neurons autoradiographically labeled, providing thus evidence that the axons of the output neurons from the homotopically transplanted olfactory bulb reconnect with the host piriform cortex.


Effects of methiothepin on changes in brain serotonin release induced by repeated administration of high doses of anorectic serotoninergic drugs. We previously observed, using in vivo microdialysis, that the potassium-evoked release of frontocortical serotonin (5-HT) is suppressed after rats receive high doses (30 mg/kg, i.p., daily for 3 days) of fluoxetine, a selective blocker of 5-HT reuptake. We now describe similar impairments in 5-HT release after repeated administration of two other 5-HT uptake blockers, zimelidine and sertraline (both at 20 mg/kg, i.p. for 3 days) as well as after dexfenfluramine (7.5 mg/kg, i.p. daily for 3 days), a drug which both releases 5-HT and blocks its reuptake. Doses of these indirect serotonin agonists were about 4-6 times the drug's ED50 in producing anorexia, a serotonin-related behavior. In addition, methiothepin (20 microM), a non-selective receptor antagonist, locally perfused through the dialysis probe 24 h after the last drug injection, enhanced K(+)-evoked release of 5-HT at serotoninergic nerve terminals markedly in control rats and slightly in rats treated with high doses of dexfenfluramine or fluoxetine. On the other hand, pretreatment with methiothepin (10 mg/kg, i.p.) one hour before each of the daily doses of fluoxetine or dexfenfluramine given for 3 days, totally prevented the decrease in basal and K(+)-evoked release of 5-HT. Finally, when methiothepin was injected systemically the day before the first of 3 daily injections of dexfenfluramine, it partially attenuated the long-term depletion of brain 5-HT and 5-HIAA levels induced by repeated administration of high doses of dexfenfluramine. These data suggest that drugs which bring about the prolonged blockade of 5-HT reuptake - such as dexfenfluramine and fluoxetine - can, by causing prolonged increases in intrasynaptic 5-HT levels as measured by in vivo microdialysis, produce receptor-mediated long-term changes in the processes controlling serotonin levels and dynamics.


Convergence of oral and extraoral information in the superior secondary gustatory nucleus of the channel catfish. Neurons within the superior secondary gustatory nucleus (nGS) of the channel catfish were examined electrophysiologically for responses to mechanical and chemical stimulation of neural peripheral receptive fields (RFs). Of the 28 single units sampled, 18 had mechanosensory RFs on the extraoral epithelium, two had RFs within the oropharyngeal cavity, and eight had RFs that included both oral and extraoral surfaces. RF sizes varied from approximately 2 cm2 on the ipsilateral lips and barbels to the whole body surface, bilaterally. No obvious correlation existed between RF pattern and recording location within the nGS. Eight of the mechanosensory nGS units also responded to amino acid taste stimuli with thresholds from micromolar to millimolar concentrations. The convergence of oral and extraoral information within the nGS determined electrophysiologically was corroborated anatomically by HRP labeling experiments. Restricted HRP injections into each of the primary gustatory nuclei of the medulla, the vagal (VL) and facial (FL) lobes, labeled fibers that appeared to terminate diffusely throughout the nGS, and injections into different portions of the nGS retrogradely labeled cells in both the FL and VL. The present electrophysiological and neuroanatomical data distinguish the convergent gustatory representation within the nGS of the catfish from the highly specific somatotopic and viscerotopic sensory maps previously identified in the FL and VL, respectively.


Effect of neurotransmitters on axoplasmic transport: acetylcholine effect on superior cervical ganglion cells. The effect of acetylcholine (ACh) on particle movements along axons of cultured superior cervical ganglion cells was analyzed with a computer-assisted video-enhanced differential interference contrast microscope system. ACh suppressed the axoplasmic transport reversibly in both anterograde and retrograde directions. A muscarinic agonist, arecoline, mimicked the ACh effect, but nicotine did not. An experiment with the Ca(2+)-indicator dye, fura-2, revealed that ACh suppressed the transport without any change of intracellular Ca2+ concentration. ACh also suppressed the axoplasmic transport in Ca(2+)-free medium. Islet-activating protein (IAP), pertussis toxin, blocked the ACh effect. These results indicate that ACh activates muscarinic receptors and suppresses fast axoplasmic transport through the activation of IAP-sensitive GTP-binding protein, irrespective of Ca2+ ions.


Barbiturate tolerance: effects on GABA-operated chloride channel function. Male ICR mice were fed powdered laboratory chow containing phenobarbital for 7 days to induce tolerance. Mice were sacrificed and brains assayed for changes in GABA-mediated chloride flux into brain membrane vesicles (microsacs). Concentration-dependent stimulation of chloride flux by GABA alone was not affected by the development of tolerance to phenobarbital. Phenobarbital potentiation of GABA-mediated chloride flux was significantly attenuated in the membranes prepared from phenobarbital-tolerant mice compared with those from pair-fed control mice. Similarly, stimulation of GABA-mediated flux by the benzodiazepine, flunitrazepam was also depressed in membranes from tolerant mice. However, the ability of ethanol and the benzodiazepine inverse agonist FG-7142 to modulate GABA-gated chloride flux was not affected by the development of phenobarbital tolerance. No significant changes in saturation [3H]diazepam binding parameters were observed. These findings suggest that there is a degree of cross-tolerance between phenobarbital and benzodiazepine agonist at the level of the GABA-operated chloride channel. Furthermore, although some reports have demonstrated behavioral cross-tolerance between ethanol and barbiturates, the present data suggest different mechanisms of tolerance development for these intoxicants at the level of the GABAA receptor chloride channel complex.


Time course of serotonergic afferent plasticity within rat spinal trigeminal nucleus following infraorbital nerve transection. High-performance liquid chromatography with electrochemical detection (HPLC-ED) and immunocytochemistry were used to examine the time course of serotonergic afferent plasticity within trigeminal subnucleus interpolaris (SpVi) following infraorbital nerve (ION) transection in adult rats. Biochemical analysis was also performed in trigeminal subnucleus caudalis (SpVc) to examine the possibility of transient lesion-induced changes in this region. No significant changes in serotonin (5-HT) or 5-hydroxyindoleacetic acid (5-HIAA) concentration, or in density of 5-HT-immunoreactive (5-HTIR) axonal varicosities were observed in either subnucleus on the lesioned side, up to 51 days following ION cut. However, at 76-79 days post-lesion, a significant increase in 5-HT concentration was again demonstrated within SpVi.


Excitotoxic lesions of the pedunculopontine tegmental nucleus of the rat. I. Comparison of the effects of various excitotoxins, with particular reference to the loss of immunohistochemically identified cholinergic neurons. The pedunculopontine tegmental nucleus (PPTg) has been shown to have cholinergic connections with the thalamus and basal ganglia. The ability of various doses of the excitotoxins (alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) (AMPA), folate, ibotenate, kainate, N-methyl-D-aspartate (NMDA), quinolinate and quisqualate to make lesions in the PPTg was examined, with particular reference to their ability to destroy cholinergic neurons identified using choline acetyltransferase (ChAT) immunohistochemistry. All of the toxins induced convulsive activity on recovery from surgical anesthesia and all except folate made lesions in the PPTg and surrounding structures. The size of the lesions was computed following examination of Cresyl violet stained sections. The largest lesions were made by kainate = AMPA greater than NMDA = ibotenate greater than quisqualate = quinolinate. All of the toxins destroyed cholinergic neurons, higher doses producing greater loss than lower. The ratio of cholinergic cell loss to general neuronal loss (assessed by Cresyl violet staining) was also computed, revealing marked differences between the toxins. Statistical analysis showed that there were significant differences between excitotoxins in terms of this ratio, but these were accounted for by the low dose of quinolinate (24 nmol) producing a significantly greater ratio of damage (12.18:1) than every other toxin. (Next highest ratio: quisqualate 60 nmol, 6.22:1.) Between the other toxins (kainate, AMPA, ibotenate, quisqualate, NMDA and the high dose of quinolinate) there were no statistically significant differences. Intense calcium deposits (stained by Alizarin red) were found frequently and often defined the borders of the lesion. Tyrosine hydroxylase immunohistochemistry revealed axons running below and into the area of lesioned tissue suggesting strongly that fibers were undamaged by the lesions. We conclude that in the PPTg, different excitotoxins make discriminably different lesions, both quantitatively and qualitatively. Unlike excitotoxic lesions in the basal forebrain quinolinate, not quisqualate, made the most selective lesions of cholinergic neurons and, unlike excitotoxic lesions in the septal nuclei, non-myelinated fibers were spared by ibotenate. The implications of these data for research into brainstem mechanisms of Parkinson's disease are discussed.


Evaluation of the neurotoxicity of N-methyl-1-(4-methoxyphenyl)-2-aminopropane (para-methoxymethamphetamine, PMMA). These studies assessed the neurotoxic potential of N-methyl-1-(4-methoxyphenyl)-2-aminopropane (para-methoxymethamphetamine; PMMA), an amphetamine analog that has surfaced in the illicit drug market. Repeated subcutaneous injections of PMMA caused lasting, dose-related reductions in regional brain concentrations of serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA), and in the density of [3H]paroxetine-labelled 5-HT uptake sites. Comparison of the neurotoxic potential of PMMA to that of para-methoxyamphetamine (PMA) and 3,4-methyl-enedioxymethamphetamine (MDMA) showed that equivalent doses of PMMA and PMA (80 mg/kg) produced comparable depletions of 5-HT, but that these depletions were not as pronounced as those induced by a lower dose of MDMA (20 mg/kg). Striatal DA was not affected on a long-term basis by any of the ring-substituted amphetamines evaluated in this study. These data suggest that PMMA, like PMA and MDMA, produces long-term (possibly neurotoxic) effects on brain serotonin neurons, but that PMMA is less potent than MDMA as a 5-HT neurotoxin. Further, they raise concern over the illicit use of PMMA since humans could be more sensitive than rodents to the 5-HT neurotoxic effects of PMMA and related drugs.


[Characteristics of monoclonal antibodies against chorionic gonadotropin receptor]. We reported the production of monoclonal antibodies (McAb) against chorionic gonadotropin (CG) receptor by fusing spleen cells of BALB/c mice immunized with purified pseudomonas maltophilia CG receptor with mouse myeloma line (SP 2/0). Four hybridoma cell lines secreting CG receptor McAbs were obtained (ED 490, DG 390, AB 890, GE 590). GE 590 is IgG 1; ED 490, DG 390, AB 890 are IgG2b. I125-HCG and ED 490, DG 390. AB 890 recognized CG receptor different antigenic determinant. Increased concentrations of GE 590 were in inverse proportion to the amount of I125-HCG binding to human ovarian tissues showing that normal ovarian tissues and ovarian malignant tumors have different HCG receptor antigenic determinant. This study indicated that these McAbs may be useful in studying the structure of CG receptor and in providing a valuable evidence for early diagnosis and treatment of ovarian malignant tumors.


Heterogeneity of IgE antibody response to reactive dye in sera from four different sensitized workers. We studied RAST and RAST inhibition tests to black GR, the most frequent sensitizer among several reactive dyes in our previous study, in order to evaluate the specificity of IgE antibodies to hapten or new antigenic determinants and the crossreactivity between two reactive dyes, black GR and orange 3R, in sera from four different sensitized workers. RAST inhibition studies with black GR-human serum albumin (HSA) conjugate discs demonstrated that black GR-HSA conjugates were the most effective inhibitors. Orange 3R-HSA conjugates, unconjugated forms of black GR and orange 3R were weak inhibitors in two patients and non-inhibitory in one patient, whereas they caused strong dose-dependent inhibitions in one patient. These results suggested that the IgE response to black GR-HSA conjugates might be heterogenous and the crossreactivity between two reactive dyes differed from one patient to another.


Wallerian degeneration in the optic nerve of the wabbler-lethal (wl/wl) mouse. Optic nerve pathology was studied in C57BL/6J wabbler-lethal (wl/wl) and control (+/+) mice at postnatal age of 4 weeks (P28). Qualitative light and ultrastructural pathology in wl/wl animals conformed to the criteria of primary axonal (Wallerian) degeneration. Most optic nerve axons in mutant animals appeared normal, as did oligodendroglia, the degree of myelination, the integrity and maturity of vascular elements, astroglia, and most myelin. Still, degenerating axons surrounded by somewhat normal myelin and axons with thickened myelin sheaths were prevalent in wl/wl mice. Dysmyelination or hypomyelination was not evident. At P28, pathology appeared more prominent in large diameter fibers. In the optic nerve of wl/wl mice, axonal degeneration preceded myelin disruption, adding this nerve to other previously reported systems undergoing Wallerian degeneration in this mutant.


Variability of crossreactivity of IgE antibodies to group I and V allergens in eight grass pollen species. Crossreactivity to Dactylis glomerata, Festuca rubra, Phleum pratense, Anthoxanthum odoratum, Secale cereale, Zea mays, and Phragmites communis of IgE antibodies against Lol p I or Lol p V was investigated by means of RAST-inhibition. Within a group of sera the degree of crossreactivity was demonstrated to be highly variable. Individual sera were not always equally crossreactive to all pollen species. A high degree of crossreactivity for Group I allergens did not necessarily implicate the same for Group V. Group I and Group V representatives were found to be present in all eight species. It was demonstrated that within this group of grass species significant quantitative and qualitative differences exist, with respect to Group I and Group V allergens. Species with a low phylogenetic affinity to Lolium perenne, like Zea mays and Phragmites communis showed a very low degree of reactivity, even when measured with the most crossreactive sera. A higher taxonomic relationship however, did not always implicate a closer antigenic resemblance. Antigenically both allergens from Zea mays are more similar to Lol p I and Lol p V, than the analogues in Secale cereale.


Regional differences in chronic neuroleptic effects on extracellular dopamine activity. The extracellular levels of dopamine (DA) and DA metabolites in the caudate-putamen (CPu) and the nucleus accumbens (NA) of rats following administration of haloperidol (HAL) decanoate and fluphenazine (FLU) decanoate for 8 months were assessed using intracranial microdialysis 1 month after final injection. Both HAL- and FLU-treated animals showed persisting plasma neuroleptic levels at time of sacrifice. Extracellular basal levels of homovanillic acid (HVA) in the CPu were significantly elevated in the FLU-treated animals, while basal levels of 3,4-dihydroxyphenylacetic acid (DOPAC) in the CPu were significantly elevated in the HAL-treated animals. Basal levels of DA and the serotonin metabolite, 5-hydroxyindoleacetic acid (5HIAA) in the CPu were not significantly different between groups. No significant between-group differences were found for basal levels of any of the analytes in the NA. Neuroleptic-treated animals showed an enhanced response to direct infusion through the dialysis probe of amphetamine (1 microM) and nomifensine (10 microM) in the CPu but not the NA. These results suggest that chronic neuroleptic treatment produces enhanced extracellular DA activity in nigrostriatal, but not mesolimbic DA pathways.


Enhancement of slow wave sleep parallel to the satiating effect of acidic fibroblast growth factor in rats. Male Wistar rats bearing both chronic cortical electrodes and ICV guide cannula were used. Acidic fibroblast growth factor (aFGF) or vehicle was injected in the lateral ventricle (20, 40, and 80 ng) while EEG and feeding patterns (via microbalance and a computerized monitor) were recorded. Forty and 80, but not 20, ng of aFGF brought about a significant reduction of feeding during 3 h and a dramatic increase of slow wave sleep during 6 h postinjection. Paradoxical sleep remained unchanged. Rectal temperature did not change. These concomitant effects of a single injection of aFGF on feeding and particularly on sleep are similar to the effect of a large meal on the same behavioral parameters.


Characterization of an O-antigen bacteriophage from Aeromonas hydrophila. A unique bacteriophage of Aeromonas hydrophila serotype O:34 was isolated, purified, and characterized. The bacterial surface receptor was shown to be the O-antigen polysaccharide component of lipopolysaccharide specific to serotype O:34, which was chemically characterized. The high molecular weight lipopolysaccharide fraction (a fraction enriched in O antigen) was fully able to inactivate bacteriophage PM1. Phage-resistant mutants of A. hydrophila O:34 were isolated and found to be specifically devoid of lipopolysaccharide O antigen. No other cell-surface molecules were involved in phage binding. The host range of bacteriophage PM1 was found to be very narrow, producing plaques only on A. hydrophila strains from serotype O:34.


Common epitopes of mammalian amelogenins at the C-terminus and possible functional roles of the corresponding domain in enamel mineralization. The present studies were undertaken to investigate the presence of common epitopes of mammalian amelogenins at the C-terminus and the possible functional importance of the conserved C-terminal domain in enamel mineralization during mammalian amelogenesis. Enamel proteins, including the intact amelogenins and their degraded polypeptides, were isolated from the secretory enamel of pig, cow, rat, and rabbit incisors. Rabbit and rat antipeptide sera, as well as rat anti-25 kD and 20 kD pig amelogenin sera, were used to identify the amelogenins among the isolated matrix proteins of each of the animal species. The antipeptide sera were developed previously (Aoba et al. [19]) using as immunogens the two synthetic peptides, C13 and C25, which correspond to the last 12 (plus Cys for KLH-conjugation) and 25 amino acid residues of pig intact amelogenin, respectively. Reactivity of the enamel proteins with each antiserum was examined by Western blot analysis. The results of immunoblotting showed that a few enamel matrix proteins in each of the mammalian species were recognized by the anti-C13 serum, specifically, pig amelogenin at 25 kD (and trace components at 27, 22, and 18 kD), cow amelogenin at 28 kD (trace components at 26, 22, 19, and 14 kD), rat amelogenins at 28 and 26 kD (and a trace component at 20 kD), and rabbit amelogenins at 24 and 21 kD (and a trace at 13 kD). The anti-C25 serum reacted additionally with pig amelogenin at 23 kD, cow amelogenin at 27 kD (a major matrix constituent), and rabbit protein at 19 kD.(ABSTRACT TRUNCATED AT 250 WORDS)


Childhood trauma and multiple personality disorder: the case of a 9-year-old girl. This paper reports the case of a 9-year-old girl, evaluated and diagnosed with multiple personality disorder over a 6 month period. Included is a detailed description of the child's presentation with historical and developmental data. A discussion of the dynamic and predisposing features of the case with references to current literature follows, along with treatment perspectives.


Construction of the physical maps of Mycoplasma hyopneumoniae and Mycoplasma flocculare and the location of rRNA genes on these maps. The genomes of Mycoplasma flocculare and Mycoplasma hyopneumoniae, two mycoplasmas of the porcine respiratory system, were studied. Based upon antigenic cross-reactivity and DNA-DNA hybridization, these species have given indication of a close genetic relationship. By using field-inversion gel electrophoresis and employing the restriction digest fragments obtained from the gels as the probes, physical maps of the genomes of the two species were constructed. Mycoplasma hyopneumoniae is similar to M. flocculare in having a single set of rRNA genes and the 5S-rRNA gene is separated from the 16S and 23S rRNA genes. Based upon the location of the rRNA genes on the physical maps in both species, the distance between the 5S and the 16S and 23S rRNA genes is at least 150 kbp. Thus, there is further evidence for the close relationship between these organisms.


Escherichia coli serotyping and disease in man and animals. Serotyping of Escherichia coli is useful, but complex, with 173 O antigens, 80 K antigens, and 56 H antigens, which can all be subdivided into partial antigens. The O, K, and H antigens can be found in nature in many of the possible combinations. The final number of E. coli serotypes is very high, 50,000-100,000 or more. The number of frequent pathogenic serotypes is, however, limited. Two main groups of such frequent serotypes are (i) serotypes from diarrhoeal disease and (ii) serotypes from extraintestinal disease. Serotypes from diarrhoeal diseases are mostly species specific, and could at present be used as epidemiological markers for bacterial clones equipped with special virulence markers, such as toxins and adhesins. Their O-antigen lipopolysaccharides may be regarded as virulence factors. These strains are not inhabitants of the normal intestine. Serotypes from extraintestinal diseases constitute a different set of clones, which are good colonizers of the intestinal tract, that under certain conditions succeed in invading host tissues. They are characterized by virulence factors different from those found in strains from diarrhoeal disease. Thus, the two groups of pathogenic E. coli are both composed of a limited number of clones for which the O:K:H serotypes are excellent, although not faultless, markers.


Pathogenicity and molecular genetics of O-specific side-chain lipopolysaccharides of Escherichia coli. Lipopolysaccharide (LPS), a glycolipid molecule found on the outer leaflet of outer membranes of gram-negative bacteria, consists of three moieties: lipid A, core oligosaccharide, and the O-specific polysaccharide chain. The O-specific side chain, which extends to the extracellular milieu, plays an important role in pathogenicity, especially during the initial stages of infection, because of its ability to interact with serum complement. In recent years, several laboratories have used recombinant DNA tools to determine, at the molecular level, the organization, expression, and regulation of genes involved in LPS biosynthesis in Salmonella and Escherichia coli. An increased understanding of the molecular aspects of the O-specific side-chain genes will shed light on the intimate details related with the formation of the O-specific side chain, its assembly onto the lipid A--core, and the translocation and insertion of the complete LPS molecule into the outer membrane. It will also contribute to the understanding of the evolution of these genes and the correlation of chemical diversity of O-specific side chains with the genetic diversity of O-specific side-chain genes. In addition, since the O-specific side chains are involved in the pathogenicity of medically important gram-negative bacteria, a basic understanding of the regulation and expression of O-specific side chain LPS genes will contribute to the field of molecular pathogenesis. This article provides an overview of the role of O-specific side chains in septicemic infections and also discusses the current status of molecular genetic studies on O-specific side-chain genes from E. coli.


Thymic hyperplasia with hemorrhage simulating recurrent Hodgkin disease after chemotherapy-induced complete remission. Thymic hyperplasia after successful treatment of malignant disease is a well-documented phenomenon, particularly in younger patients with testicular carcinoma. However, it has been reported only sporadically in association with Hodgkin disease. The authors report the first instance of an adult with nodular sclerosing Hodgkin disease who had thymic hyperplasia develop after clinical and radiographic complete remission (CR) with chemotherapy alone. In addition to the current case, 20 other instances of thymic hyperplasia have been culled from the literature and analyzed. The median age of these 21 patients was 23 years of age (range, 8-40 years of age). Thymic hyperplasia was noted after treatment was initiated or completed in all but two patients; and all did well, with one exception: a 17 year-old boy who was inadvertently treated for relapsed Hodgkin disease and who died of fibrinous pneumonitis and nocardia, with pathologic CR noted at autopsy. Thymic hyperplasia in association with Hodgkin disease appears to be a favorable prognostic phenomenon, restricted to younger patients in the first modal peak of Hodgkin incidence. Prospective studies have yet to be performed.


Successful treatment of a primary endodermal sinus tumor of the liver. A 27-year-old woman had a large hepatic tumor and a markedly increased serum alpha-fetoprotein (AFP) level. A diagnosis of endodermal sinus tumor was made after a needle biopsy was performed on the liver. Clinical and radiologic examinations did not show an alternative primary site. Treatment with cisplatin, etoposide, and bleomycin was started, but, after three cycles, was changed to cisplatin, vincristine, methotrexate, bleomycin, dactinomycin, cyclophosphamide, and etoposide because the serum AFP level was decreasing too slowly. After additional chemotherapy was given, the patient was well but had an increased AFP level and a large residual mass in the liver. A right hemihepatectomy was performed, but no viable tumor was present. The patient is alive and disease-free 5 years later. Thus, AFP levels may be misleading in the presence of large necrotic tumors. The authors stress the need to make a diagnosis of these rare tumors early because aggressive treatment with combination chemotherapy may result in cure.


Prognostic significance of changes in prostate-specific markers after endocrine treatment of stage D2 prostatic cancer. The prognostic value was determined of prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP) measured before and after endocrine treatment in 57 patients with newly diagnosed Stage D2 prostatic cancer. Therapy included orchiectomy or administration of luteinizing hormone releasing hormone analogues or an antiandrogen. The absolute pretreatment PSA (elevated in 100% of patients) but not PAP (abnormal in 93%) predicted disease progression (P < 0.0011), i.e., a poor response to therapy. Fifty-three patients responded to androgen deprivation with a decrease in PSA level. This declined to normal at 3 and 6 months in 25% of patients. Forty-nine percent had a greater than 90% decrease in their PSA level. By 1 year, 58% of patients had progressive disease. Both the nadir PSA level and the percent decline from the pretreatment level at 3 and 6 months predicted the progression-free interval (P < 0.001). Patients with a 90% or greater decline in PSA had a prolonged progression-free survival. Serial PAP levels were similarly prognostic. It was concluded that PSA was better than PAP in evaluating patients before and after androgen-deprivation therapy. The nadir level of both markers was an important tool to predict progression-free survival in patients with metastatic prostatic cancer.


A phase II study of recombinant human alpha-interferon in advanced hormone-refractory prostate cancer. To determine the efficacy of recombinant human leukocyte alpha-interferon (IFL-RA) in advanced hormone-refractory prostate cancer, the authors treated 40 patients with IFL-RA administered intramuscularly at a dose of 10 x 10(6) U/m2 three times weekly. Toxicity was substantial and necessitated at least a 50% dose reduction in all but five patients during the first 1-2 months of therapy. No responses were observed in patients with bone metastases, but complete and partial regression of nodal disease were observed in two patients with extraosseous disease (overall response rate, 5%; 95% confidence interval, 0.64-17.75%). The authors conclude that IFL-RA cannot be recommended at this dose and schedule in patients with advanced prostate cancer, but additional study of its use in patients with nodal disease may be warranted.


Multiple cancers in the prostate. Morphologic features of clinically recognized versus incidental tumors. Multiple independent tumors were identified in specimens from 117 of 234 prostatectomies for clinical adenocarcinoma; there were 266 incidental cancers in these 117 prostates. The clinically detected carcinoma was the largest (or only) tumor in all 202 Stage B cases. However, among 32 Stage A cases (detection by transurethral resection), there were 8 prostates in which an incidental tumor was larger than the clinically manifest cancer. These were all small tumors except for two incidental cancers with a volume greater than 2cm3; roughly 80% of incidental carcinomas were smaller than 0.5 cm3, whereas fewer than 20% of manifest tumors were smaller than 0.5 cm3. Comparison with a series of cancers found incidentally at cystoprostatectomy for bladder cancer showed the same volume distribution as incidental (smaller) carcinomas in patients with prostate cancer. This distribution was thought to reflect the volume distribution of prostate cancer in the general population older than 50 years of age. It was concluded that additional incidental tumors are common in patients with prostate cancer, but their sum of volumes is seldom as large as the clinical cancer volume.


An immunohistochemical study of epithelial membrane antigen, cytokeratin, and vimentin in papillary thyroid carcinoma. Recognition of lethal and favorable prognostic types. Immunoreactivity for epithelial membrane antigen (EMA), cytokeratin, and vimentin was investigated in 15 papillary thyroid carcinomas (PTC) with distant metastases, 25 PTC without distant metastases, and 34 occult PTC without distant metastases that were found incidentally at autopsy. More than 50% of the tumor cells were positive for EMA in 7 (47%) of 15 PTC with distant metastases, 0 (0%) of 25 PTC without distant metastases, and 1 (3%) of 34 occult PTC. The incidence of EMA positivity in PTC with distant metastases was significantly different from that of both PTC without distant metastases and occult PTC (P < 0.001). Cytokeratin reactivity was similar in the three groups, and almost all PTC stained strongly for cytokeratin. Concerning vimentin positivity, there were no significant differences in three groups; however, PTC with distant metastases tended to stain more weakly or focally than PTC without distant metastases or occult PTC. These results suggest that EMA reactivity may be a useful factor for anticipating the individual risk of distant metastasis or death from PTC at the time of initial surgical treatment.


Surgical resection of solitary metastases after chemotherapy in patients with nonseminomatous germ cell tumors and elevated serum tumor markers. Chemorefractory metastatic germ cell tumors and elevated tumor markers generally indicate inoperable disease. Solitary metastases were resected in 15 patients who had a nonseminomatous germ cell tumor and an elevated alpha-fetoprotein (AFP) and/or human chorionic gonadotropin (HCG) serum level after treatment with cisplatin-based chemotherapy. Patients underwent resection for a residual mass after chemotherapy or for a new solitary metastasis after achieving a complete response (CR) to salvage chemotherapy. Seven patients were disease-free after surgical resection alone. All five patients with an elevated HCG level had a relapse after surgery compared with 3 of 10 patients with only an elevated AFP level. Only 4 of 10 patients with a retroperitoneal metastasis had a relapse after surgery compared with 4 of 5 patients with visceral disease. Eleven of 15 patients overall were disease-free after surgery and subsequent chemotherapy after a relapse. Surgical resection of a solitary metastasis despite elevated serum tumor markers should be considered in patients who have not had a durable CR to cisplatin-based chemotherapy.


Co-ordinate expression of c-fos, p53 and cytokeratin genes during the alteration of growth of human hepatoma cells. mRNA levels measured by reverse transcription and polymerase chain reaction. Teleocidin, a tumor promoter, inhibited the proliferation, enhanced cytokeratin assembly and increased the type III procollagen production of PLC/PRF/5 hepatoma cells. Teleocidin transiently increased the levels of c-fos and p53 mRNAs measured by reverse transcription and polymerase chain reaction. This was followed by a reduction of c-myc mRNA and an increase of cytokeratin mRNA. The level of p120 mRNA was not remarkably altered. Sequential alterations of the expression of c-fos, p53, c-myc and cytokeratin genes induced by teleocidin may be responsible for the morphological and functional changes of hepatoma cells induced by this tumor promoter.


Activation of human CD4+CD45RA+ T cells by chrysotile asbestos in vitro. Chrysotile asbestos stimulates T lymphocyte subsets. Cell surface CD4 or CD45RA expression in peripheral blood mononuclear cells (PBMC) was downregulated after incubation with chrysotile asbestos in vitro temporarily. The percentage of CD4+CD45RA+ cells and mean fluorescence intensity in CD4 or CD45RA decreased after incubation with asbestos and returned to the original level after 24 h of incubation, which suggests that chrysotile asbestos activates CD4+CD45RA+ cells. No change was observed in CD29 expression. An increased percentage of IL-2R positive cells and an elevated intracellular Ca++ level were also indicative of the activation of PBMC by chrysotile asbestos.


P-glycoprotein expression during tumor progression in the rat liver. P-Glycoprotein (Pgp) has been shown to mediate multidrug resistance in tumor cell lines. Overexpression of Pgp has been detected in clinical cancer samples of many histological types. The basis and biological significance of such increases in Pgp expression are not well understood. In this study, the expression of Pgp during stepwise progression to rat liver cancer was examined to investigate the possible role of Pgp in carcinogenesis. An immunohistochemical technique was used to detect Pgp at the single-cell level, in a large number of liver nodules, hepatocellular carcinoma, and in distant metastases of the carcinomas. The results showed that distinct changes in Pgp expression occurred during stepwise liver carcinogenesis and that these changes were closely associated with the microscopic anatomy of the lesions. In contrast to gamma-glutamyl transpeptidase and glutathione S-transferase-7.7, whose expression appeared to correlate with the early steps of liver carcinogenesis, Pgp expression was higher in the large hyperplastic nodules and in hepatocellular carcinomas than in the early microscopic lesions. A particularly striking finding was the consistent expression of Pgp in the lung metastases. These findings suggested that Pgp was associated with a more progressed malignant phenotype in liver carcinogenesis.


Loss of epithelial markers and acquisition of vimentin expression in adriamycin- and vinblastine-resistant human breast cancer cell lines. We have previously observed that breast cancer cell lines could exhibit either epithelial or fibroblastic phenotypes as reflected by their morphologies and intermediate filament protein expression (C. L. Sommers, D. Walker-Jones, S. E. Heckford, P. Worland, E. Valverius, R. Clark, M. Stampfer, and E. P. Gelmann, Cancer Res., 49:4258-4263, 1989). Fibroblastoid, vimentin-expressing breast cancer cell lines are more invasive in vitro and in vivo (E. W. Thompson, S. Paik, N. Brunner, C. L. Sommers, G. Zugmaier, R. Clarke, T. B. Shima, J. Torri, S. Donahue, M. E. Lippman, G. R. Martin, and R. B. Dickson, J. Cell. Physiol., 150: 534-544, 1992). We hypothesized that a breast cancer cell with an epithelial phenotype could undergo a transition to a fibroblastic phenotype, possibly resulting in more invasive capacity. We now show that two Adriamycin-resistant MCF-7 cell lines and a vinblastine-resistant ZR-75-B cell line have undergone such a transition. Adriamycin-resistant MCF-7 cells express vimentin, have diminished keratin 19 expression, have lost cell adhesion molecule uvomorulin expression, and have reduced formation of desmosomes and tight junctions as determined by reduced immunodetection of their components desmoplakins I and II and zonula occludens (ZO)-1. Other MCF-7 cell lines selected for resistance to vinblastine and to Adriamycin and verapamil did not have these characteristics, indicating that drug selection does not invariably cause these phenotypic changes. In addition, to determine if vimentin expression in MCF-7 cells alone could manifest a fibroblastic phenotype, we transfected the full-length human vimentin complementary DNA into MCF-7 cells. Although vimentin expression was achieved in MCF-7 cells, it did not affect the phenotype of the cells in terms of the distribution of keratins, desmoplakins I and II, ZO-1, or uvomorulin or in terms of in vitro invasiveness. We conclude that vimentin expression is a marker for a fibroblastic and invasive phenotype in breast cancer cells but does not by itself give rise to this phenotype.


Insulin-like growth factor binding proteins in human breast cancer tissue. Breast tumor cell lines have been shown to secrete at least five distinct insulin-like growth factor (IGF) binding proteins (IGFBP), the secretion being related to the estrogen receptor (ER) content. In this study we investigated IGFBP mRNA expression and IGFBP content in relation to ER content in human breast tumors. Tissue specimens from 47 breast cancers were studied. In five cases the adjacent histologically normal tissue was also analyzed. IGFBP content in tissue homogenates was studied by Western ligand blot analysis, using [125I] IGF-I as a label, and IGFBP mRNA expression by reverse transcriptase polymerase chain reaction. The results show that human breast tumors express mRNAs encoding IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4 and IGFBP-5. The pattern of IGFBPs in different tumors varies. No correlation exists between ER content and IGFBPs with molecular weights 24,000 Mr, 28,000 Mr, 34,000 Mr or 43,000 Mr, whereas the 49,000 Mr IGFBP was more abundant in ER negative tumors (P less than 0.05). The IGFBP content was significantly (P less than 0.05) higher in five tumors than in their adjacent normal tissues suggesting that increased content of IGFBPs is a feature typically associated with the malignant transformation of breast tissue.


Product of the Steel locus can replace leukemic cell interaction. Mutations in the Steel locus, encoding a growth factor (Steel factor or SF) or c-kit, the gene encoding its receptor, result in severe anemia in the mouse. In the present study, we have addressed the mechanism of synergistic growth activation, at the cellular level, by SF and GM-CSF using the blast cells of acute myeloblastic leukemia (AML blasts). Our data indicate that SF drastically alleviates the requirement in cell interaction for blast colony formation in most of the samples tested. Analysis of cultures performed in the presence of SF and GM-CSF at different cell concentrations, ranging from 1,000 to 20,000 cells, suggested a single limiting element, i.e., the blast clonogenic cell, while 2 or more limiting elements were found in cultures stimulated with GM-CSF alone, suggesting interacting cell populations. The presence of membrane-bound SF was detected by immunofluorescence, suggesting the possibility that secreted or membrane-bound SF may, at least in part, contribute to the density-dependent growth of AML blasts. In all samples tested, SF appears to increase the responsiveness of AML blasts to GM-CSF, as demonstrated by a 3-fold decrease of GM-CSF half efficient concentration on addition of SF to the cultures. Exposure of AML blasts to SF did not affect GM-CSF receptor expression, suggesting that this increase in GM-CSF responsiveness is likely to occur at the postreceptor level. Interestingly, 2 of 15 AML samples surveyed did not respond to SF, and were both of the myelomonocytic or monocytic subtype, classified as M4 and M5, respectively.


Flow cytometric detection of drugs altering the DNA methylation pattern. We have developed a model system for assessing the demethylating potential of external agents. Disruption in the DNA methylation pattern was evaluated at the translational level of the Escherichia coli beta-galactosidase coding gene (lacZ). We have constructed a clonal cell line (A4/4 cells) derived from the adenovirus-transformed human embryonic kidney 293 strain. The A4/4 cells contain the E. coli lacZ gene under the control of the mouse metallothionein 1 promoter which is down-regulated by a natural DNA methylation pattern. Furthermore, the lacZ transcription is also regulated by the E. coli lac operator/repressor system and by mouse metallothionein 1 metal responsiveness offering a wide range in lacZ expression. In this system, the beta-galactosidase activity was only recovered in the presence of a demethylating agent such as 5-azacytidine. The demethylating potential of 5-azacytidine, 5-aza 2'-deoxycytidine and sodium butyrate was rapidly assessed by a flow cytometric method using fluorescein di-beta-D galactopyranoside as a fluorescent probe. A tremendous induction of lacZ expression was triggered by these drugs. Analysis of cell cycles showed little disruptions with 5-azacytidine and sodium butyrate, but an important blockage in the S-phase following 5-aza 2'-deoxycytidine treatment was observed. This approach allows a rapid identification and study of environmental demethylating agents.