Effect of eating liquorice on the renin-angiotensin aldosterone axis in normal subjects. The effect of confectionery liquorice on electrolyte status and the renin-angiotensin-aldosterone (RAA) axis was studied in 14 healthy volunteers. They ate liquorice in daily doses of 100 g or 200 g (equivalent to 0-7-1-4 g glycyrrhizinic acid) for one to four weeks. Plasma potassium concentrations fell by over 0-3 mmol/l in 11 people, including four who had to be withdrawn from the study because of hypokalaemia. One or more values of the RAA axis, especially plasma renin activity and urinary aldosterone concentrations, were considerably depressed in all subjects. These results show that potentially serious metabolic effects may occur in some people who eat modest amounts of liquorice daily for less than a week.


An in vitro analysis of metal electrodes for use in the neural environment. The results of evaluation of the corrosion response of Pt, Au, Rh, Ir, Pt-10% Ir and Pt-10% Rh electrodes subjected to bipolar current pulses of 0.1 and 1.0 A/geom. cm2 in a simulated neural environment are reported. The criteria for evaluation include the extent and morphology of material removal and corrosion product formation. The chemical changes on the electrode surface and in the test electrolyte are also evaluated. Modified potentiographic methods were developed which monitor interface conditions of the electrodes during chronic, long-term biphasic current passage. Under the test conditions established by the neural model, evaluation of the corrosion response of the candidate electrode materials shows that Rh was moderately damaged at 1.0 A/geom. cm2, while the remaining test electrodes were severely damaged at this current density. While Rh, Ir and Au exhibit corrosion resistance at 0.1 A/geom. cm2, damage to all of the test materials preclude their use in any long-term chronic neural prosthetic implant under the simulated stimulation parameters of the tests. The relative corrosion resistance of the test electrode materials is related to the adherence and coherence of the corrosion product produced on the electrode surface during the first few hundred hours of stimulation. Analysis of the voltage response of the electrodes to current passage provides a means of monitoring the nucleation and growth of the corrosion product.


Responses of single neurons to electrical stimulation of the surface of the visual cortex. We have recorded from single neurons in the visual cortex of the cat while stimulating the cortical surface with the same type of electrodes and the same parameters of stimulation which have produced phosphenes in conscious man. Parameters of stimulation which will permit excitation of single cortical neurons with little risk of producing afterdischarges are described. The patterns of excitation of single neurons during surface stimulation and the effects of stimulation on neuroglial cells have also been studied. Surface stimulation also produced marked alterations in cortical excitability as tested with visually presented stimuli. Mechanisms by which single neurons are excited by surface stimulation are also considered.


The visual cortex during chronic stimulation. The lateral and suprasylvian gyris of cats have been stimulated anodically by surface electrodes for a total of 40 h by biphasic square waves. Peak currents were +/-6.0 mA, O to peak. Control electrodes were never stimulated. Electrode materials were low-temperature isotropic carbon and metals (Rh, Pt, Au). Reductions in tissue impedance quickly followed the onset of stimulation. The impedance change was related to stimulus current and to the incidence of motor seizures but not clearly to the electrode material. Utilizing concentric electrodes, the impedance change was not localized to the stimulating electrode - tissue interface. No protection from stimulation-associated impedance modifications was provided by the systemic administration of a material of high osmolarity (Mannitol) but the usual impedance decrease was not seen after systemic administration of a glucocorticoid.


Rapid dendritic atrophy following deafferentation: an EM morphometric analysis. The nucleus laminaris (NL), a third-order brain stem auditory nucleus in birds, receives afferents to its dorsal dendrites from the ipsilateral nucleus magnocellularis (NM), while the ventral dendrites of NL neurons are innervated by axons from the contralateral NM via the crossed dorsal cochlear tract (CTrX). The CTrx was transected in young chickens and, 96 h later, NL was examined for cytological changes. A morphometric analysis of electron micrographs from lesioned and shamd axon terminals were localized almost entirely to the ventral neuropil region of NL. (2) The volume density of dendrite in the ventral region of NL of lesioned animals was reduced by 85% compared to both the dorsal dendritic region of the same animals and the ventral dendritic region of sham-operated control animals. (3) The frequency with which primary dendrites were encountered in the ventral neuropil of lesioned animals was 81% lower than sham-operated controls. (4) Frequently, there was an apparent reduction in the amount of rough endoplasmic reticulum, Golgi apparatus and cytoplasmic granularity in the deafferented part of the NL neurons. (5) The data indicate that the cytological integrity of dendritic processes cna be specifically, profoundly and very rapidly compromised by removing a significant proportion of their afferents, suggesting that a tonic influence may be exerted by the presynaptic terminals.


Effects of posterior neocortical lesions on wavelength, light/dark and stripe orientation discrimination in ground squirrels. Thirteen-lined ground squirrels (Citellus tridecemlineatus) were trained on three two-choice visual discrimination problems: light/dark, color and stripe orientation. After posterior neocortical lesions in one or two stages, they were tested on all three discriminations. The results demonstrate that animals with large posterior neocortical lesions which produced retrograde changes throughout the dorsal lateral geniculate (LGNd) were capable of light/dark and wavelength discrimination. These animals were not able to discriminate stripe orientation. It is proposed that wavelength discrimination depends on extrageniculostriate mechanisms in posterior neodecorticates of this species.


Analysis of sleep-wakefulness rhythms in male rats after suprachiasmatic nucleus lesions and ocular enucleation. To determine quantitatively characteristics of sleep-wakefulness rhythms in male albino rats, computer analysis of long term polygraphic records (24 h/day) of cortical EEG activity, neck EMG and EOG taken from 23 rats under 12:12 light-dark schedule was performed. After bilateral suprachiasmatic nucleus (SCN) lesions, the circadian rhythm in sleep-wakefulness was completely eliminated, although no attenuation or even slight enhancement of the ultradian rhythms with 2-4 h pweiodicity was observed. After enucleation of both eyes, the circadian rhythm was free-running with a phase shift in the range from --12 to +22 min/day in 6 rats. A gradual decrease of the spectral value of the circadian rhythm and inverse enhancement of the ultradian rhythms with 4--7 h periodicity (predominantly 6 h in 4 out of 6 rats) were also shown. In the spectral diagram, the appearance of paradoxical sleep (PS) paralleled slow-wave sleep (SWS), in the cases of the circadian rhythm and ultradian rhythms with 4--7 h periodicity. Behaviorally blind rats with bilateral primary optic tract (POT) lesions maintained the circadian rhythm in sleep-wakefulness entrained to the environmental light-dark cycle. Power spectral analysis showed no characteristic difference from normal rats. Based on these data, the role of the SCN as a pacemaker of endogenous circadian rhythm in sleep-wakefulness is discussed.


Morphine, naloxone and the responses of medial thalamic neurones of the cat. Administered electrophoretically morphine depressed the firing of medial thalamic neurones. This effect was not antagonized by naloxone which itself was a depressant. A long lasting increase in sensitivity to L-glutamate followed ejection of both morphine and naloxone. Intravenous morphine, 0.5-1.5 mg/kg, had inconstant effects on spontaneous firing that evoked by electrical stimulation of the forepaws but in 7 of 10 experiments reduced the sensitivity of neurones to L-glutamate. This effect was reversed in all cases by intravenous naloxone, 0.3 mg/kg


Genetic variation in brain L-glutamate decarboxylase activity from two inbred strains of mice. Two inbred strains of mice, C57BL/6Bg and DBA/1Bg, were compared for genetic varaition in brain L-glutamate decarboxylase (GAD) activity. Although no large difference was found between the strains in whole brain GAD activity at adult age (27--45 days postnatally), regional examination revealed a difference in GAD activity in the cerebral cortex (15% higher in DBA); subcellular examination revealed a difference in synaptosomal fraction (21% higher in DBA). When GAD activity was measured in synaptosomal fractions prepared from dissected cerebral cortex, DBA was 34% higher than C57BL. In addition, differences in GAD activity between the two strains could be observed even in the whole brain (10--15% higher in DBA) during earlier development (15--23 days postnatally). These data indicate that at adult age, genetic difference in GAD activity between the two strains exists mainly in nerve terminals of the cerebral cortex. It is postulated that the difference may be due to a genetically mediated mechanism regulating GAD in the presynaptic terminals of GABA neurons of the cerebral cortex.


Characteristics of dopamine and beta-adrenergic sensitive adenylate cyclases in the frontal cerebral cortex of the rat. Comparative effects of neuroleptics on frontal cortex and striatal dopamine sensitive adenylate cyclases. Homogenates of frontal cerebral cortex of the rat were prepared from microdiscs punched out in areas rich in dopaminergic terminals. Under optimal assay conditions, dopamine (10-4 M) stimulated an adenylate cyclase present in these homogenates by 80-100%. This stimulation reached 200% when microdiscs were punched out from the medial part of the frontal cerebral cortex, adjacent to the forceps minor. Dopamine interacted with an homogeneous population of receptor sites which had an apparent affinity (KD) of 3.8 +/- 0.9 x 10-6 M (N = 4). The dopamine receptor was blocked by fluphenazine and phentolamine but had no affinity for pindolol, propranolol or L-isoproterenol. The affinities of several neuroleptics having different chemical structures were simultaneously determined on striatal and on frontal cerebral cortex dopamine sensitive adenylate cyclases. Fluphenazine was more potent in blocking the striatal than the frontal cerebral cortex dopaminergic receptors. In contrast, in all experiments, haloperidol had an higher affinity for the cerebral frontal cortex than for the striatal dopaminergic receptors. Thus, haloperidol was less effective than fluphenazine in blocking the striatal dopaminergic receptors, and equally potent than fluphenazine in inhibiting the frontal cerebral cortex dopamine sensitive adenylate cyclase. Chlorpromazine, thioridazine and clozapine had the same affinity for the two dopaminergic adenylate cyclase systems. L-isoproterenol interacted with an homogeneous population of beta-adrenergic receptor sites (KD = 3 +/- 2 X 10-7 M; N = 4) coupled with an adenylate cyclase distince from the dopamine sensitive adenylate cyclase. This beta-receptor had no affinity for dopamine or fluphenazine but was blocked by propranolol or pindolol. L-Norepinephrine was shown to stimulate both the dopamine (KD = 1.8 +/- 1 X 10-5 M; N = 4) and the beta-adrenergic (KD = 8 +/- 3 X 10-7 M; N = 4) sensitive adenylate cyclases. Thus, the L-norepinephrine effect was totally blocked in the combined presence of fluphenazine and pindolol.


An enzymatically stable peptide with activity in the central nervous system: its penetration through the blood-CSF barrier. Vasopressin, other neurohpophyseal peptides and certain of their fragments can affect central nervous system function. Cyclo(Leu-Gly), formally derived from the C-terminal dipeptide of oxytocin, is also active in memory processes. This molecule, radioactively labeled for this study, was found to be resistant to enzymatic degradation both in vivo and in vitro. Furthermore, after its intravenous application in cat, it easily penetrated into cerebrospinal fluid. This indicates that cyclo(Leu-Gly) can readily pass from blood into the central nervous system to exert it's central pharmacological actions. Thus, labeled cyclo(Leu-Gly) is a valuable model for studying the distribution and localization of peptides which have activity in the central nervous system.


Subcellular distribution of rat brain cortex high-affinity, sodium-dependent, glycine transport sites. The subcellular distribution of the membrane components, present in rat brain cortex homogenates, that interact with glycine in the presence of sodium ions was studied. The distribution in the primary fractions, as per cent of total binding in the homogenate, was: P1 ('nuclear'), 58%; P2 (large granule), 39%; P3 (microsomal), 2%9 Of the subfractions obtained by centrifuging P1 in a linear 0.32--1.5 M sucrose gradient, only the lighter fraction (P1-III) formed by large myelin fragments was enriched in specific binding activity with respect to P1. The pellet formed by purified nuclei had negligible binding, and fractions of intermediate density had a lower activity than P1. Transient exposure of P1-III to 1.5 M sucrose did not diminish its binding ability. Similarly, in the subfractions obtained by centrifuging P1 in a discontinuous sucrose gradient, only the least dense one, P1-A, that is formed exclusively by large myelin fragments, was enriched with respect to P1. The electron microscopy of these fractions is presented. The P2 subfractions, obtained in a linear 2--18% Ficoll gradient, had the following sodium-dependent activity (counts/min/mg protein, fractions being in the order of decreasing density): pellet, 0; P2-I, O; P2-II, 450; P2-III, 1770; P2-IV, 4130; unfractionated P2, 880; P2-IV, the least dense fraction being composed mainly of myelin. With P2 subfractions obtained in a discontinuous sucrose gradient (0.32, 0.8 and 1.2 M sucrose layers), it was also found that sodium-dependent glycine binding was only enriched, with respect to P2, in the myelin fraction P2-A. Glycine binding to purified brain cortex myelin was also found to be very high, while binding to non-myelin membranes, obtained during the purification procedure, was only 0--7% of that seen with myelin. These results suggest that high-affinity glycine binding is located in myelin proper, and possibly also in some other glial plasma membranes, but not in nuclei, mitochondria, endoplasmic reticulum or synaptosomes. The relevance of these findings for interpreting previous reports on high-affinity glycine transport in the central nervous system is analyzed.


Electrophysiological and horseradish peroxidase studies of precerebellar afferents to the nucleus interpositus anterior. I. Climbing fiber system. The input to the nucleus interpositus anterior (NIA) of the cat from the inferior olive (IO) was studied by stimulating the IO while recording intracellularly from the NIA, and by the retrograde transport of horseradish peroxidase (HRP). Stimulation of the IO evoked monosynaptic EPSPs in NIA neurons. The cells labeled in the IO following electrophoretic and pressure injection of HRP into NIA were located in the rostral parts of the dorsal and medial accessory olive. Stimulation of the IO also polysynaptically evoked an IPSP and a late disinhibitory depolarization. Data were presented which indicated that these potentials were mediated by the Purkinje cells of the cerebellar cortex.


Electrophysiological and horseradish peroxidase studies of precerebellar afferents to the nucleus interpositus anterior. II. Mossy fiber system. Inputs to the nucleus interpositus anterior (NIA) of the cat from precerebellar nuclei which are thought to give rise to mossy fibers were studied using electrophysiological and anatomical techniques. Stimulation of one of these precerebellar nuclei, the lateral reticular nucleus (LRN) evoked monosynaptic EPSPs in NIA neurons. These EPSPs were followed by polysynaptic IPSPs and late depolarization mediated by the response of the cerebellar cortex. Similar responses were occasionally seen following stimulation of the brachium pontis (BP). When horseradish peroxidase was injected into the NIA, labeled cells were found in the magnocellular and parvicellular LRN, the external cuneate nucleus (ECN), the pontine nuclei and the perihypoglossal nuclei. There was no evidence for a direct projection of the nucleus reticularis pontis to the NIA. It was suggested that most of the tonic excitation of NIA neurons is provided by the LRN and ECN.


Habenular and other midbrain raphe afferents demonstrated by a modified retrograde tracing technique. Afferents to th midbrain dorsal and median raphe nuclei in the rat were studied by means of the horseradish peroxidase (HRP) retrograde transport method. The HRP was given by means of a modified iontophoretic delivery technique. This technique permitted an efficient and localized deposition of a high concentration of HRP into the raphe nuclei. Afferents to the raphe as determined by this method could be categorized into 2 classes; those exclusively to the raphe and those also positive for adjacent reticular formation. The most striking afferent area to the raphe, both in terms of selectivity and density, was the lateral habenula. This result is in accord with previous studies using degeneration methods which indicate an habenular projection to the raphe area. There were afferents exclusively positive for the dorsal raphe nucleus emanating from the nucleus of the solitary tract. Most other raphe afferent areas were also positive for the reticular formation (e.g;, prefrontal cortex, medial forebrain bundle, preoptic nuclei, and reticular formation). The existence of a major afferent system from the lateral habenula to the midbrain raphe is consistent with the concept of a "dorsal pathway" which might be responsible for relaying information from forebrain limbic structures to the "midbrain limbic areas".


The relation between end-tidal CO2 and discharge patterns of sympathetic preganglionic neurons. In 11 Nembutal-anesthetized, vagotomized, thoracotomized, paralyzed and artificially ventilated cats, the electrical activity of 32 single sympathetic preganglionic neurons (SPNs), dissected from the cervical nerve was recorded at various end-tidal CO2 levels, together with the activity of the phrenic nerve. Seven of these neurons were insensitive to CO2 changes, within a range of end-tidal CO2 values from 1;0 to 10;0%. All 7 had a background firing pattern without respiratory modulation, even at the highest CO2 levels tested, i.e., had the same firing frequency in both phases of the phrenic nerve activity cycle. Seventeen units were silent at low CO2 levels, began to discharge at particular CO2 levels (on the average, at 2.3% CO2) and increased their firing frequency (on the average, by 0.9 spikes/sec/% CO2) as end-tidal CO2 was raised above the threshold level. Their background discharge pattern was characterized by firing only in the inspiratory phase of the phrenic nerve activity cycle. Three units had firing which was CO2-independent within a range of low CO2 concentrations and which increased as CO2 concentration was increased above this range. These units fired throughout the phrenic nerve activity cycle but had their peak frequency in inspiration. Five units had a firing frequency which was highest at low CO2 and which decreased with increasing CO2 levels. These units had their peak frequency in expiration. These results show that the output of this SPN population is strongly influenced by CO2 within the range of concentrations tested. The finding that sensitivity to CO2 changes is a property only of SPNs with respiratory-modulated firing pattern suggests that the CO2-dependent input is relayed to these SPNs via the respiratory center; A comparison of data obtained under hypocapnic conditions with data obtained in previous studies in normocapnic cats with mid-cervical spinal cord transections suggests that brain stem inspiratory neurons represent a major excitatory input to this SPN pool.


Central actions of histamine: microelectrophoretic studies. Responses of neurones in different regions of the rat and cat brain to micro-electrophoretically applied histamine and some related substances including antagonists are reported. Histamine excites most neurones in the hypothalamus but depresses the vast majority of neurones in other structures. Depressant actions of histamine and some inhibitions in the cortex and hippocampus of the rat are antagonized by metiamide, an H2-receptor antagonist. The results lend strong support to the idea of histaminergic transmission in the mammalian brain.


The occurrence and seasonal variation of gigartinine and L-citrullinyl-L-arginine in Chondrus crispus Stackh. Gigartinine, 5-(3-amidinoureido)-2-aminovaleric acid, and L-citrullinyl-L-arginine were islated from aqueous extracts of Chondrus crispus (Rhodophyceae). Their identifications were confirmed by chemical procedures, and 1H and 13C nuclear magnetic resonance and infrared spectroscopic methods. Citrullinylarginine, gigartinine, taurine, citrulline, and glutamic acid were the predominant free amino compounds. Citrullinylarginine showed the most pronounced change in concentration. This occurred during the winter months when it reached a maximum (in March) of 58 mumol/g fresh weight, a value 10 times greater than that of any of the free amino acids and equal to 50% of the total organic nitrogen in the plant; All of these compounds were depleted to a minimum of about 1 mumol/g fresh weight in October. Both gigartinine and citrullinylarginine were detected in Ahnfeltia plicata, Gracilaria sp. Petrocelis middendorfii, Polyides rotundus, Polysiphonia lanosa, and Rhodomela confervoides.


Cytoplasmic compartmentation of glucose 6-phosphate. When rat diaphragm, intact or homogenized, was incubated with [14C]glucose, the initial specific activity of the fructose 6-phosphate was higher than that of glucose 6-phosphate (glc-6-P). This relationship was also found when the cytosol was incubated with the labeled glucose. The early time course of production of glc-6-P from glucose and its removal provide evidence that newly synthesized glc-6-P does not equilibrate rapidly with the total pool of this metabolite and is preferentially removed by isomerization. It is concluded that glc-6-P is functionally compartmented within the cytoplasm.


Morphine-like activity of sheep beta-lipotropin and of its tryptic fragments. Sheep beta-lipotropin (beta-LPH) (sequence 1-91) was selectively cleaved with trypsin after blocking the epsilon-amino groups of lysine with citraconic anhydride. The resulting peptides were purified by a combination of cation-exchange chromatography and high-voltage electrophoresis. The purified fragments were then tested for their morphine-like activity in the mouse vas deferens bioassay. The active peptides were 61-91 and 61-80 were about as active as the synthetic methionine-enkephalin, and in turn these were about 100 times more active than beta-LPH itself. The inhibition of electrically stimulated mouse vas deferens by these peptides is reversed by naloxone, and suggests a competitive character of interaction. It is thus concluded that the active core for the morphine like activity in the mouse vas deferens bioassay is the fragment 61-65 of beta-LPH.


Steroid beta-D-glucosidase in steer liver and kidney. Homogenates of liver and kidney tissue from young steers had beta-D-glucosidase (EC 3.2.1.21) activity toward 17alpha-estradiol-3-glucoside, 17beta-estradiol-3-glucoside, 17alpha-estradiol-17-glucoside, and deoxycorticosterone-21-glucoside. The activity towards the phenolic 3-glucosides was largely present in the 100 000 X g supernatant, while that towards 17alpha-estradiol-17-glucoside was concentrated in the microsomes. The use of beef liver preparations for the hydrolysis of steroid 'glucuronide' fractions could result in hydrolysis of other steroid glycosides which might be present.


Radioactive conversion products of intramuscularly injected [4-14C]formononetin including sulfates in the urine of hens. The phytoestrogen formononetin was injected intramuscularly as [4-14C]formononetin into two adult hens. Radioactive materials in the urine for the succeeding 14 days (hen 1) or 16 days (hen 2) were fractionated on DEAE-Sephadex-25 columns by elution with a gradient of NaCl; the four major fractions thus separated were examined by solvent partition, thin-layer chromatography, and enzymic cleavage. The following seven radioactive components were identified in the urine, the average proportions of each being given in terms of percentage of total 14C recovered from the urine: [14C]formononetin (4.3%); [14C]diaidzein (11.4%); [14C]equol (6.8%); [14C]daidzein monosulfate (30.4%); [14C]equol monosulfate (5.8%); [14C]diadzein disulfate (19.8%); and [14C]equol disulfate (6.5%). Small proportions of sulfates of unidentified radioactive phenols were present. Tests for presence of glucosiduronates of 14C-labelled material gave negative results. Radioactive formononetin sulfate was not detected in the urine of either hen.


Synthesis and solution conformation of [Trp1, Val5]-angiotensin II. [Trp1, Val5]-Angiotensin II was synthesized by the solid-phase method and purified by partition chromatography on Sephadex G-25 and by ion-exchange chromatography on Sephadex SP-25. Relative to [Val5]-angiotensin II, the analog displayed 36% smooth muscle activity in a preparation of the superior mesenteric artery. Conformational aspects of the analog were revealed by fluorescence techniques. The fluorescence emission maximum at 350 nm suggests a completely aqueous environment for the tryptophanyl residue in [Trp1, Val5]-angiotensin II. Singlet-singlet resonance energy transfer between Tyr in position 4 and Trp in position 1 was evaluated for a calculation of the intramolecular distance between these two residues on the basis of the Förster equation. From the relative increase of tryptophan fluorescence a transfer efficiency of greater 0.9 was obtained, which is compatible with the observed complete quenching of tyrosine fluorescence in the analog. The computed average intramolecular distance of less than 8 A precludes an extended conformation for the N-terminal sequence encompassing residues 1 through 4 at neutral pH and suggests the existence of a loop in this part of the molecule. The results are discussed in relation to the various models proposed for the solution conformation of angiotensin II.


Electron-microscopic visualization of transcriptionally active and less active chromatin fractions from the rat ventral prostate and their content of histones. The content of histones in transcriptionally active (euchromatin-like (E)) and transcriptionally less active (heterochromatin-like (H)) fractions from the rat ventral prostate was determined. The absolute amount of total histone was less in the E fraction, which contained relatively more stained histone F2b and F3 and less histone F1 than either the H fraction or unfractionated chromatin. The phosphotungstic-acid-stained E fraction was composed predominantly of shorter (50-500 nm) and thinner (5-15 nm) structures. In some preparations, pieces estimated to be 500-1000 + nm in length were present. Spherical structures, from 10 to 20 nm in diameter, some of which were present as 'dimers,' trimers,' or higher-ordered ensembles were seen in the shadow-case E fraction. The stained or shadow-cast H fraction contained longer (200-2000 + nm) and thicker (congruent to 20 nm) fragments with a more complex substructure. When the shadow-cast H fraction was sufficiently dispersed, it had a nodular or 'beaded' appearance.


A study on the reaction mechanism of adenosine 5'-phosphosulfate reductase from Thiobacillus thioparus, an iron-sulfur flavoprotein. The reaction mechanism of adenosine 5'-phosphosulfate (APS) reductase (EC 1.8.99.2) from Thiobacillus thioparus was studied using difference spectrum and stopped-flow techniques. The enzyme-bound FAD was rapidly reduced by sulfite with a first order rate constant of 97.1 s-1. The addition of AMP induced further spectral changes in the reduced enzyme which were consistent with the oxidation of FADH2 to the red (anionic) semiquinone FADH-) and the concomitant reduction of nonheme iron to the ferrous state. Superoxide dismutase (EC 1.15.1.1) or anaerobiosis inhibited the reduction of cytochrome c by the enzyme only to the extent of 25-35%, indicating the existence of a direct reduction of cytochrome c by the enzyme without involving O2-. the activity of enzyme with cytochrome c was inhibited by increasing the potassium phosphate concentration, the inhibition being more pronounced with horse heart cytochrome c than with Candida krusei cytochrome c.


Wheat embryo ribonucleates. VII. Rapid, efficient and selective formation of 5S-18S and 5.8S-26S hybrids in an aqueous solution of the four ribosomal polynucleotides, and the results of a search for the corresponding hybrids in wheat embryo ribosomes. (1) If wheat embryo 5S and 5.8S rRNA are differentially labelled, it can be shown that there is highly selective association of 5S [14C]RNA with 18S rRNA, and of 5.8S [3H]RNA with 26S rRNA when a solution (0.3 M NaCl) that contains approximately equimolar amounts of the four ribosomal polynucleotides is heated briefly (3 min) at 60 degrees C. (2) Comparison of Tm values and melting profiles for laboratory-prepared and natural 5.8S-26S rRNA hybrids suggests that restoration of the natural union between 5.8S and 26 rRNA can be achieved with facility and fidelity in the laboratory. (3) Union between 5.8S rRNA remains intact when wheat embryo ribosomes are disintegrated either by digestion with pronase or by treatment with sodium dodecyl sulphate, but the same treatments release 5S and 18S rRNA as freely migrating electrophoretic components. (4) Intact 18S and 26S rRNA can be prepared from small and large subunits, respectively, when wheat embryo ribosomes are dissociated by treatment with 0.5 M KCl. (5) Incidental to the principal investigation, it has been shown that, even after storage for more than 6 years at -70 degrees C, commercial supplies of roller-milled wheat germ yield S23 extracts that are very active in the cell-free translation of globin mRNA. (6) The physicochemical and possible biochemical significance of various types of intermolecular complexing between pairs of ribosomal polynucleotides is a subject of discussion.


beta-Glucosidase: microbial production and effect on enzymatic hydrolysis of cellulose. The enzymatic conversion of cellulose is catalyzed by a multiple enzyme system. The Trichoderma enzyme system has been studied extensively and has insufficient beta-glucosidase (EC 3.2.1.21) activity for the practical saccharification of celluose. The black aspergilli (A. niger and A. phoenicis) were superior producers of beta-glucosidase and a method for production of this enzyme in liquid culture is presented. When Trichoderma cellulase preparations are supplemented with beta-glucosidase from Aspergillus during practical saccharifications, glucose is the predominant product and the rate of saccharification is significantly increased. The stimulatory effect of beta-glucosidase appears to be due to the removal of inhibitory levels of cellobiose.


Requirements for the rapid conversion of macroconidia of Fusarium sulphureum to chlamydospores. Chlamydospores of Fusarium sulphureum were formed within 72 h by incubating macroconidia at 37 degrees C in stationary culture in a medium containing mannitol and inorganic salts. The conversion process was dependent on inoculum level, temperature, aeration, and the presence of an external source of nitrogen and carbon. Chlamydospore formation was not inhibited by an uncoupler of oxidative phosphorylation, 2,4-dinitrophenol, or by inhibitors of bacterial cell wall synthesis; however, it was affected by inhibitors of respiration and of protein and chitin synthesis.


Ontogeny of lipid bodies in the endoplasmic reticulum of Fusarium sulphureum. The ontogeny of lipid bodies during the differentiation of conidial cells to chlamydospores in Fusarium sulphureum (F. sambucinum f. 6) is described. The lipid bodies are first seen as electron-transparent areas within dilated cisternae of the endoplasmic reticulum (ER). One or more such cisternae may be formed within a strand of ER. The intracisternal lipid bodies are bounded by a unit membrane derived from the double membrane of the ER. The dilated cisternae increase in size and remain electron-transparent; however, some electron-dense granular material is present in the lipid bodies. The limiting unit membrane is visible throughout development of the lipid body; however, in mature chlamydospores the membrane is obscured by a broad band of electron-dense material which may provide protection to the stored lipids during adverse environmental conditions. The system of lipid body development in F. sulphureum is discussed in relation to those described for higher plants.


Characterization of an immunogenic fraction of Pasteurella multocida culture filtrates. An immunogenic fraction (IF) of Pasteurella multocida strain P-1059 was separated from culture filtrate by Sephadex gel filtration. Additional fractionation of IF with aqueous ether resulted in the glycoprotein-like preparation (GLP) while extraction with aqueous phenol provided the lipopolysaccharide-like preparation (LPP). The unextracted IF contained carbohydrate, protein, and lipid; the GLP contained carbohydrate and protein; and the LPP contained carbohydrate and lipid. The GLP was maximally protective for mice against homologous challenge, and was medially toxic in rabbit skin when compared to the other culture-filtrate preparations; the LPP was maximally toxic in rabbit skin, and was least protective for mice; and the unextracted IF was medially protective for mice, and was least toxic in rabbit skin.


Effects of proteolytic enzymes on invertase secretion in sphaeroplasts of Saccharomyces: inhibition by trypsin. Exogenously added trypsin arrested invertase secretion by sphacroplasts of Saccharomyces strain 1016. The mechanism of inhibition is presumed due to attack on plasma membrane protein(s). Gross membrane damage by trypsin was not apparent, as evidence by the absence of leakage of intracellular alkaline phosphatase, after trypsin treatment. Trypsin treatment did induce an increased sensitivity to lysis, observed only when changes in osmotic pressure were made and fresh glucose added. While synthesis of invertase was eventually inhibited by trypsin, a greater than twofold increase in internal invertase was observed, due to complete inhibition of secretion. This is the first report of the uncoupling of synthesis and secretion in yeast.


Manganese-mediated morphogenesis in Penicillium claviforme and Penicillium clavigerum. The colonial morphology and growth of Penicillium claviforme and P. clavigerum in manganese-deficient, defined medium, were examined with and without manganese supplementation. In unsupplemented medium rendered free of manganese through treatment with a chelating resin, P. claviforme grew but did not produce coremia. Similar results were obtained for P. clavigerum on media devoid of added manganese. However, when these media were supplemented with optimal concentrations of manganese both fungi produced coremia. The presence of manganese in various colony sections oed in the conidia.


Small cells in pure cultures of Agromyces ramosus and in natural soil. Previously, transmission electron-microscopic observations of thin-sectioned and frozen-etched preparations of microorganisms concentrated directly from soil showed many cells that were considerably smaller than the cells found in pure cultures isolated from soil. The present study shows that pure cultures of the soil bacterium Agromyces ramosus can produce cells as small as those observed in natural soil. Growth on a nutritionally limiting medium and the presence of an agar surface contributed to the occurrence of these small cells.


Hyperlipoproteinemia and cerebral strokes. Determination of lipoprotein patterns and of major plasma lipids was carried out in 171 patients with strokes. Of these, 22 had hyperlipoproteinemia (HLIPR) by paper electrophoresis and by elevation of principal plasma lipids (either cholesterol over 300 mg/100 ml or triglycerides over 200 mg/100 ml or both these components). More than two-thirds of the patients had at least one close relative with elevated blood lipids. Using criteria of the World Health Organization, these patients were classified as follows: 5 had HLIPR type IIa, 8 had HLIPR type IIb, 3 had HLIPR type III and 6 had HLIPR type IV. Phospholipids showed relatively little change from the values of normal controls. The numerical distribution of patients with stroke and HLIPR into the four different types corresponds quite well with the approximate frequency of these types of HLIPR in the general population. Thus, this study does not indicate that the patients with a particular type of HLIPR are at a greater risk to have a stroke than those belonging to other types.


Mechanisms of the "low-flow" state during resuscitation of the totally ischemic brain. Resuscitation of the brain following total circulatory arrest may be impeded by difficulty in establishing cerebral tissue perfusion, a postischemic "low-flow"state. We have confirmed this hypothesis in a rat model of total cerebral ischemia and have demonstrated marked imporvement in post-ischemic brain tissue perfusion following epinephrine injection. This is mainly due to the systemic vascular effects of epinephrine, resulting in improved central arotic pressure and cerebral perfusion pressure. Hyperkalemic induced vasoconstriction has also been postulated as a cause of the "low-flow/. We have, therefore, investigated the in vitro effects of increasing potassium ion concentration on cerebrovascular smooth muscle strips. Large arteries constrict, while small arteries dilate in response to hyperkalemia. The net effect on cerebral blood flow remains unsettled. Our research to date suggests that resuscitation of the totally ischemic brain, in animal models at least, is enhanced by epinephrine, mainly via its effects on central aortic pressure.


An autoradiographic study of muscular dystrophy, motor neuron disease and Charcot-Marie-Tooth disease. The autoradiographic findings using tritiated leucine are described in muscle biopsy material from five patients with progressive muscular dystrophy (P.M.D.), three with motor neuron disease (M.N.D.) and four with Charcot-Marie-Tooth disease (C.M.T.). In progressive muscular dystrophy there is a marked increase in uptake of leucine into cytoplasmic proteins and precursors, and reduced incorporation into structural protein. In Charcot-Marie Tooth disease muscle there is a significantly increased uptake into cytoplasmic elements and a normal uptake into structural protein. In motor neuron disease the uptake into cytoplasmic elements appears normal but is reduced into structural proteins. The abnormal uptake in C.M.T. could be explained as a product of regenerative efforts associated with reinnervation. However, the abnormal uptake may represent the primary effects of gene action in the muscle, as seems probable in progressive muscular dystrophy.


Response recovery cycles in the visual cortex and superior colliculus following conditioning "ON" and "OFF" stimulation in the rabbit. The responsiveness of visual cortex (VC) and superior colliculus (SC) was simultaneously compared following conditioning "ON" or "OFF" stimulation, in the rabbit. Average evoked responses were recorded simultaneously from the visual cortex ans superior colliculus. "ON" or "OFF" steps constituted the conditioning stimuli whereas the test stimulus consisted of optic nerve stimulation. All evoked responses exhibited a reversal of their polarity when the electrode was moved in the dorsoventral direction (Negative-Positive in the SC, Positive-Negative in the VC). This assured the somato-dentritic origin of the potentials. The results showed that responsiveness in both structures was significantly higher following an "OFF" simulus than after an "ON" step. Collicular responsiveness was higher than in the VC when the same conditioning stimulus was applied. The spatial distribution of the source of "OFF" responses was circumscribed to the ventral part of the superficial layer of the superior colliculus. These results suggest specific properties associated with the brightening and dimming systems.


Wicket spikes: clinical correlates of a previously undescribed EEG pattern. From an analysis of the electroencephalograms of 4,458 patients who underwent recording during both wakefulness ans sleep, through the years 1969 to 1975, wicket spikes-- recorded in 39 patients-- may be described as follows: They were found during both wakefulness ans sleep, almost exclusively in adults. Their cardinal feature is a changing mode of occurrence through any single recording: from intermittent trains of more or less sustained, arciform, discharges resembling mu rhythm, to sporadic, urinary, single spikes. When occurring singly, wicket spikes can be mistaken for anterior or middle temporal spikes, since they predominate in eith er area, and since they share with them other characteristics such as amplitude (60 to 210 microvolts), polarity (surface negative) duration, and configuration. Wicket spikes should not be considered interictal abnormalities; they do nor correlate with epilepsy or with any particular symptom complex.


Transfer and interference in amygdaloid kindling in cats. Examination of the effects of primary site kindling upon the development of secondary site amygdaloid kindling in cats showed that the former exerts powerful positive as well as negative after effects upon the latter. The positive after effect appears to be directed towards establishing a linkage between the secondary site and the Stage 6 generalized convulsive mechanism. The negative after effects were strikingly similar to those observed in animals with forebrain bisection subjected to amygdaloid kindling, suggesting that it is primarily directed towards functional accessibility of the cerebral structures interconnected by the forebrain commissure. The similarity of the findings in animals with anterior neocortical lesions supports such an assumption. The transient nature of the negative after effects was indicated by the development of a 'normal' clinical ictal pattern from the secondary site when it occurred spontaneously. Secondary site kindling exerted similar but less prominent negative after effects when the primary site was re-tested. Stage 6 seizure was elicited upon the first trial, however.


Regional cerebral blood flow statistical patterns and psychological performance in multi-infarct dementia and Alzheimer's disease. Multivariate data analysis statistical procedures were applied to identify regional cerebral blood flow (rCBF) patterns in patients with multi-infarct dementia (M.I.D.) and Alzheimer's disease (A.D.) when compared to a control group (C.). A fronto-temporal-parietal pattern was identified for the A.D. group. The M.I.D. group demonstrated a predominant temporal parietal pattern. A discriminant function analysis classified 96% of the A.D. and C. patients correctly. One hundred per cent hit rate was obtained in discriminating between M.I.D. and C. Discrimination of A.D. and M.I.D. obtained 82% diagnostic accuracy. When the three groups were compared the hit rate was 93%. Mean rCBF and mean relative weight of the gray matter (Wg) differences were found when each dementia group was compared with C.M.I.D. and A.D. did not differ in mean rCBF but significant differences were found for Wg, with M.I.D. having greater reduction. The performance on standardized psychological tests of two independent samples of A.D. and M.I.D. were compared. The Memory Quotient from the Wechsler Memory Scale was consistently more imparied for the A.D. group. No correlation between degree of psychological impairment and rCBF or Wg reduction was found.


[Obstructive parietal hemodissection of the intracranial vessels, a particular form of dissecting aneurysm]. A six year old girl complained of sudden severe headache, became hemiplegic and unconscious. A right carotid arteriogram revealed an obstruction of the right anterior cerebral artery and many sulvian branches. Death occurred four days later. At autopsy, a recent softening of nearly all the right middle cerebral arterial territory was found. Thrombus filled the sylvian artery and its main branches. Histologic examination of the vessel walls showed a dissecting infiltration of blood between the internal elastic lamina and the media. This particular form of dissecting aneurysm, occurring in young subjects, in the absence of atherosclerosis, high blood pressure and idiopathic medial necrosis, represents a distinct medial necrosis, represents a distinct nosologic entity that has been called "Obstructive parietal hemodissection of intracranial vessels." The pathogenesis of the disease is unknown: trauma has been mentioned, also congenital defects in the elastic lamina or other morphologic abnormalities of that lamina.


Myopathy associated with amyloid angiopathy. A 38-year-old patient with the clinical picture of a progressive myopathy resembling limb girdle muscular dystrophy is presented. Muscle biopsy showed amyloid deposits in the walls of small endomysial blood vessels. There was no clinical or physiological evidence of peripheral nerve involvement, no plasma cell dyscrasia and no generalized amyloidosis. There was no muscle fiber hypertrophy, inflammation or neurogenic change. Theere was no response to steroid therapy. The etiopathogenesis of this amyloid angiopathy is undetermined. The extensive vessel involvement with amyloid deposition and the absence of changes indicative of muscular dystropyy or inflammatory myopathy leads us to favor an ischemic basis for this patient's myopathy.


Antimicrobial drug use in three Canadian general hospitals. Total amounts of antimicrobial drugs used to treat inpatients during 1975 were calculated for three Canadian general hospitals, one of them the principal teaching hospital of a medical school. Use of drugs was compared with that reported for Boston City Hospital during periods when antimicrobial therapy was and was not supervised by infectious disease consultants. Ampicillin, tetracyclines, cephalosporins, erythromycin and aminoglycosides for prophylactic oral administration were used excessively in the three hospitals. The degree of overuse was comparable to that at Boston City Hospital during years when drug use was uncontrolled. Overuse or improper choice of antimicrobial drug decreases the quality of patient care and increases its cost. More rigorous education is needed for both medical students and practising physicians in the rational use of antimicrobial drugs. Informal consultation with an infectious disease unit should be required before certain overly popular or toxic antibiotics are administered to hospitalized patients.


Rational and irrational use of antibiotics in a Canadian teaching hospital. Parenteral therapy with gentamicin, cloxacillin, ampicillin and cephalothin was surveyed on a surgical, a gynecologic and medical ward of a teaching hospital. During a 3-month period 219 patients (12.9% of the total number admitted to the three wards) received at least one of the four antibiotics parenterally. Ampicillin and gentamicin were used most frequently on the three wards when the indication for therapy was either infection or empirical use. Cephalothin was used most frequently for prophylaxis in the gynecologic and surgical patients; no medical patient received this drug. Overall, therapy was assessed to be irrational in 42.0, 50.0 and 12.0% of the surgical, gynecologic and medical patients, respectively. Prophylaxis was the indication for therapy in 76.9 and 86.8% of the surgical and gynecologic patients, respectively, for whom the therapy was assessed to be irrational.


Adverse reactions to 1975 bivalent influenza vaccine in children. A questionnaire survey was undertaken to investigate incidence and types of reactions to a bivalent, zonally purified, killed influenza vaccine administered to 155 children aged 1 to 18 years in 1975. Local or systemic reactions or both occurred in 42 children (27%). However, these reactions were not considered serious and should not constitute a major drawback to future influenza programs in children under 3 years of age.


Absence of cell-mediated immunity to rubella virus 5 years after rubella vaccination. The long-term effectiveness of rubella vaccination in childhood is particularly important because the ultimate goal of immunization is the prevention of infection during pregnancy. Of 25 healthy children tested 4 to 5 years after rubella vaccination, 19 showed no evidence of cell-mediated immunity (CM) to rubella virus despite the presence of hemagglutination-inhibition or complement-fixation antibodies or both. Twenty-two of 25 seropositive, naturally infected young adults showed evidence of CMI. These results indicate that fetuses of women who have been vaccinated against rubella may not be protected against damage by wild rubella infection during the pregnancy, when CMI is physiologically depressed.


Guide to the management of rubella problems. Despite the availability of rubella vaccine the incidence of the congenital rubella syndrome has been increasing in certain regions of Canada. Perhaps this is not surprising in view of the known irregular cyclic activity of rubella virus in a community and the fact that the percentage of seropositive women of childbearing age has not changed appreciable since the vaccine was introduced. Clearly vaccine is not being administered to sufficient numbers of women at risk. Until a much higher percentage of women of childbearing age possess rubella antibody, the costly problem of congenital rubella syndrome is likely to be with us. Common rubella problems relate to four categories: the exposed pregnant woman, laboratory diagnosis, the infant with suspected congenital rubella and the vaccine. One of the most common questions about the vaccine is the following: Can recently vaccinated individuals disseminate vaccine and infect seronegative contact? The answer is No.


Nutrition survey of schoolchildren in greater Winnipeg. I. Descriptive and anthropometric data. By sequential random numbering 10 schools in greater Winnipeg were selected for a nutrition survey. Interviews were conducted with 201 grade 3 children and 182 grade 6 children for whom parental consent was obtained. Of these, 48 in grade 3 and 51 in grade 6 were studied in further detail. There were no differences in descriptive data between the general and detailed groups or among the 10 schools. Most fathers were skilled or unskilled labourers and about 50% of the mothers were homemakers without outside employment; parental occupation did not influence eating patterns. Breakfast was the meal most often missed; 8% of the 383 children had come to school without breakfast. Since many children in grade 3 had prepared their own breakfast and since there was a relative lack of physical activity, school health programs should incorporate more than nutritional supplements and nutrition education. On the basis of body weight and height the nutritional status of the 99 children studied in detail was judged to be generally satisfactory; according to the Boston standards the boys were heavy and tall, and the girls were normal in weight but short.


Nutrition survey of schoolchildren in greater Winnipeg. II. Dietary intake and biochemical assessment. The total dietary intake of energy and of individual nutrients of 99 grades 3 and 6 children from 10 greater Winnipeg shcools were generally comparable to those reported by Nutrition Canada for the Manitoba and national samples, although the percentile distributions of total caloric intake and dietary intake of vitamin A for the Winnipeg children tended to be lower. The median daily intake of protein was 212% of the Canadian Dietary Standard and most came from animal sources. Dietary fat was largely from animal sources as well. Hemoglobin concentrations were marginally low in four children, and urinary riboflavin:creatinine ratios were low in six children. There was no biochemical evidence of thiamin deficiency. The results suggest a need for change in dietary patterns and for education in nutrition, including relative nutrient/cost benefits. A well planned school snack program with an education component is a medium by which change could be introduced. This should preferable be part of a total school health program.


Congenital hip disease in a Cree-Ojibwa population: a retrospective study. Retrospective study of data from annual surveys and hospital records over 23 years confirmed the 1950 report of a high prevalence of congenital hip disease (CHD) in the Cree-Ojibwa population of Island Lake, Man. Annual ascertainment rates ranged from 35 to 600 cases per 1000 live births; 5-year rates for dislocation or subluxation were the highest reported for any population. The minimally declining rates of CHD may reflect upgrading in criteria for hip abnormality as well as decreasing isolation and increasing outbreeding of the population. The preponderance of females (female:male ratio, 1.90:1) was low compared with that found in other studies. For all diagnoses bilateral hip involvement exceeded unilateral in frequency; laterality differences were not significant when the sexes were studied separately. Function in everyday activities was impaired little.


Lead poisoning from lead-soldered electric kettles. Lead poisoning occurred in two infants, causing lead encephalopathy in one but no symptoms in the other. Both infants had been fed a formula prepared with water boiled in a lead-soldered electric kettle. The diagnosis was suggested by dense metaphyseal bands in radiographs, illustrating the importance of careful examination of "routine" radiographs, particularly in asymptomatic infants.


Massive intestinal infarction in young women: complication of use of oral contraceptives? Massive intestinal infarction due to occlusion of the celiac, superior mesenteric and inferior mesenteric arteries occurred in two young women, one of who subsequently died. Both were smokers. They had ingested oral contraceptives for 5 and 8 years, respectively, but this therapy could not be proven to be a causative factor in their ischemic bowel disease; although such an association is uncommon, it should be considered in young women with abdominal pain.


Enterobacter agglomerans: the clinically important plant pathogen. During a 5-month period Enterobacter agglomerans, now described as a member of the phytopathogenic genus Erwinia, was isolated from 13 patients in a general hospital; in 1 patient it was isolated from two sites. In six instances the organism was the sole pathogen isolated, in two instances it may have contributed to infection and in the remaining instances it was probably a transient saprophyte. The strains showed some variation in biochemical reactions but were similar in colonial morphology and were consistently sensitive to several antibiotics. Although this organism is prevalent in the general environment and usually relatively benign, it does have a potential for nosocomial infection.


Preliminary report of the lung cancer detection program in New York. The early lung cancer detection program in New York is described, and preliminary prevalence rate data are reported. Cigarette-smoking men over the age of 45, who are considered at high risk, are encouraged to enroll in the program and, to date, 6,612 have done so. These men receive PA and lateral chest x-rays annually, while a randomly selected sub-population of about half also have sputum cytology every four months. A controlled study of sputum cytology as an adjunct to the chest x-ray is now in progress. There were 15 cases of late lung cancer (pathologic stages II and III) detected at the initial examination, and 11 early cases (pathologic stages O and I). In the population of 3,387 men who had x-rays alone there were three early lung cancers detected; in the population of 3,225 men who had both examinations four early cancers were detected by x-ray and four by cytology. In addition, three cases of larynx cancer were detected by cytology. The early lung cancers detected by x-ray were peripheral and most were bronchiolar or adenocarcinoma; the early cancers detected by cytology were central epidermoid carcinoma of major bronchi. No early carcinomas were detected by both techniques. These findings suggest that a combination of chest x-rays and sputum cytology can be more effective than either one alone in detecting early lung cancer.


Suspected pulmonary metastases: correlation of chest x-ray, whole lung tomograms, and operative findings. Chest x-rays, whole lung tomograms, and operative findings were correlated in 152 patients with an extrathoracic malignancy who underwent 182 thoracotomies for the evaluation of pulmonary nodules. Among 25 patients with a normal chest x-ray but one or more pulmonary nodules demonstrated by whole lung tomography, 19 had metastatic disease documented at thoracotomy. Of 64 patients with unilateral nodules seen on conventional chest x-ray, 10 of 32 with sarcoma, 2 of 7 with melanoma, but only 2 of 25 with carcinoma had bilateral nodules demonstrated by tomography. At thoracotomy, 8 of 38 patients in whom a solitary nodule was present on both chest x-ray and on tomogram had multiple nodules. Similarly, 35 of 48 patients with more nodules seen on the tomograms than on the chest x-ray had even more extensive metastatic disease demonstrated by operation. Among patients who had resection of all metastatic pulmonary nodules, 50% of those with carcinoma and 32% of those with sarcoma who underwent resection of a solitary nodule approached that of patients with carcinoma. These findings suggest that whole lung tomograms contribute to the determination of the extent of pulmonary metastasis in patients with solid malignancies, even in the presence of a negative chest x-ray. The improved long term survival achieved by resection of these metastases may be partially attributed to this screening procedure.


Paragangliomas of the head and neck region: a clinical study of 69 patients. Over a 38-year-period 73 paragangliomas from the head and neck region were seen at Memorial Hospital. These occurred in 69 patients. There were 44 carotid body, 13 vagal body, eight jugulo-tympanic, and three nasal paragangliomas. In addition, one each arose in the orbit, larynx, and area of the aortic arch. Sixty-two patients were surgically treated while three received irradiation only and four were observed but not treated. Follow-up was obtained for 94% of the patients. There were six postoperative deaths, five of which occurred before 1945. Of those treated surgically, 37 had no evidence of recurrent tumor while the paragangliomas recurred locally in 11. All patients in the radiation and untreated groups had persistent tumor. Malignant behavior with death due to tumor was observed in five cases (four carotid body and one vagal body paraganglioma). Using modern techniques carotid body and vagal body paragangliomas can usually be managed by surgery alone. Other paragangliomas of the head and neck due to their anatomic location are sometimes best treated by a combination of surgery and radiation or only by radiation therapy.


Psychophysiologic aspects of cancer: the James Ewing lecture. Clinicians have long been aware of the influence of the neuro-endocrine axis and the action of the hypothalmus on the humoral immune response in the origin and course of cancer. Little attention has been paid, however, to the psychophysiologic aspects of cancer. These psycho-social effects may be related to hypothalamic activity, the autonomic nervous system, and neuro-endocrine activity. More attention should be paid to the manipulation of the psyche in the prevention and management of cancer.


The use of CEA as an early indicator for gastrointestinal tumor recurrence and second-look procedures. Since 1972 plasma CEA levels of 25 cancer patients have been assayed to evaluate the reliability of CEA as an early indicator of recurrent gastrointestinal cancer. Identification of significant elevations in CEA levels required definition of exactly what a given value meant. Intraassay and interassay accuracy was determined and graphed as a CEA NOMOGRAM, which measures the observed CEA level against the 95% confidence limits for that observation and thus can be used to identify statistically significant increases. A statistically significant rise above a baseline value established by the NOMOGRAM proved to be a correct indicator of tumor recurrence in 22 (88%) of 25 patients who underwent second-look intraabdominal operations (22 colorectal, 2 gastric, and 1 pancreatic). In each case, other accepted procedures, such as liver enzymes, scans, and x-rays, were nondiagnostic. Of the 22 patients with proved tumor recurrence, 16 (73%) had distant metastases and 6 (27%) had localized tumors. One patient remains tumor-free three years after second-look operation and has had no significant change in CEA levels. More frequent serial CEA determinations combined with sound clinical judgment should facilitate earlier detection of recurrent gastrointestinal cancer.


An approach for the treatment of penile carcinoma. One hundred cases were studied in detail using lymphangiograms (LAG), anatomic dissections, and/or microscopic evaluation. LAG performed via the dorsal lymphatics of the penis demonstrate the existence of specific lymph node center, the so-called sentinel lymph node (SLN). This appears to be the primary site of metastases from penile carcinoma. The SLN is visualized radiographically, on the antero-posterior view, at the junction of the femoral head and the ascending ramus of the pubis. Anatomically, the SLN is part of the lymphatic system around the superficial epigastric vein. Forty-six SLN biopsies were performed with 15 positive for metastatic disease. In these 15 patients, an inguinofemoroiliac dissection was performed; in 12 cases there was no involvement of other lymph nodes. Lymphatic channels draining into the iliac lymph nodes without first draining into the sentinel lymph node were never demonstrated, nor were the inguinal-femoral lymph nodes involved in the absence of SLN involvement. On this bases, we recommend preliminary bilateral SLN biopsy to be followed by inguinofemoroiliac dissection when biopsy of the SLN biopsy to be followed by inguinofemoroiliac dissection when biopsy of the SLN is positive. When biopsy of the SLN is negative for metastatic disease, no further surgical therapy is immediately indicated. With negative SLN, 5-year survival was 90%. When SLN alone was involved, 5-year survival was 70%. Five-year survival was 50% with both SLN and other inguinal nodes involved. When iliac metastases were also present, 3-year survival was 20%.


Economic status and survival of cancer patients. The relation of economic status to survival was studied for 39 kinds of cancer representing all types for which 60 or more indigent patients were seen in University of Iowa Hospital for primary care during the years 1940-1969. For every type the indigent patients had poorer survival than non-indigent patients. Quality of care would be eliminated as a major variable since a second group of "ward" patients of higher economic status was available for comparison and the differences were substantially greater between the two groups of teaching patients than between the "clinic pay" and "private" patients. Age differences and differences in stage of disease accounted for less than half of the survival deficits in the indigents. The two important problems were high mortality from causes other than cancer and excess cancer mortality not accounted for by stage differences, particularly among patients who should have had 5-year survival rates between 40 and 70%. In these patients cancer recurred more often and earlier among the indigent. We postulate host differences associated with poverty that could also account for much of the observed Black-White differences as well as some international differences in cancer survival rates.


Physical and psychologic readjustment after mastectomy: an evaluation of Memorial Hospitals' PMRG program. Memorial Hospital in New York City, the clinical section of the Memorial Sloan-Kettering Cancer Center, has developed a Post-Mastectomy Rehabilitation Group (PMRG) Program. The PMRG Program is comprised of a series of structured exercise, information, discussion, and group therapy sessions conducted by a social worker, nurse, physical therapist, and a volunteer from Reach to Recovery. The specific purposes of the program are to help a patient to begin to regain functional use of her arm and shoulder on the affected side, and to adapt functionally, psychologically, and emotionally to the loss of her breast and to the diagnosis of cancer in the shortest time possible. Approximately 1,700 women who underwent mastectomy participated in the program between 1970 and 1974. Some results and conclusions are: the patient's age and type of operation are related to physical recovery, resumption of normal activities, return to work, and emotional stress; 84% (661 of 790 patients) resumed normal activities within the four-month postoperative period; 74% (237 of 317 patients) returned to work (full time) within three months of their mastectomies; 13% (52 of 406 patients) experienced moderate or severe emotional stress; activity per se (any activity, doing something, keeping busy) affects readjustment; the apparent disadvantage of greater trauma with more extensive surgery may be more than compensated for by the relatively young age of that population, and the relatively less physical trauma and injury, resulting from simple mastectomies may be overshadowed by the patient's age and general physical condition; 98% (284 of 291 patients) reported the PMRG program helpful.


Intravenous Corynebacterium parvum: an adjunct to chemotherapy for resistant advanced ovarian cancer. This is a prelimianry report of an effort to treat women with advanced (Stage III and IV) ovarian cancer who had progressive disease in spite of previous surgery, chemotherapy and/or radiation by a program of reductive surgery, intensive immune stimulation and combination chemotherapy. An initial laparotomy was done where possible to reduce tumor burden, and then all patients were given intravenous corynebacterium parvum (C.P.) in escalating doses over a 10- to 14-day period. Cyclic chemotherapy with Cytoxan, adriamycin and 5-fluorouracil (CAF) was started and repeated monthly. Maintenance subcutaneous C.P. was given weekly. All patients had frequent follow-up clinical and laboratory examination. Immune function was measured by skin tests and in vitro tests prior to treatment and periodically during therapy. Two-thirds of the patients had depressed DNCB and PHA stimulation responses prior to treatment, and almost all had severely depressed lymphocyte counts. Thirty-nine patients entered the program. Exploratory laparotomy was done in 16 patients and in eight, successful tumor reduction was accomplished. Eleven patients received intravenous C. Parvum and all expired before receiving chemotherapy. Four patients received C. Parvum and less than three cycles of CAF; all expired within 2 months. Twenty-four patients received C. Parvum and less than or equal to three cycles of CAF. Four patients had complete regression of measurable disease and were living free of disease 9-12 months after starting CAF. Eight patients had more than 50% regression for a minimum of 3 months, and five were living with disease (LWD) 5-11 months. Five patients had 25% to 50% regression and three were LWD 4-8 months. Seven patients had no regression and all expired within 4 months. Of eight patients who had successful reductive surgery prior to treatment, three were free of disease, median of 10 months, and five had partial responses and were living with disease, a median of 9 months. Although pre-treatment immune function was better in the patients who had a good response to CP and CAF (10 of 12 were DNCB+) vs that in patients with a poor response (4 of 12 were DNCB+) immune function was not significantly improved during therapy. The initial treatment results in this program are encouraging and suggest that this approach may be useful in patients with earlier disease.


Carcinoma of the breast: analysis of total lymph node involvement versus level of metastasis. The histologic presence of axillary lymph node metastasis is the most commonly used indicator or prognosis for patients with operable breast cancer. The record of 385 patients treated by 380 radical mastectomies between the years 1944 and 1972 were reviewed to clarify this. The axillary nodes recovered were evenly distributed. The median number of positive lymph nodes at each level was two; 50% had involvement of only one level. The number of lymph nodes identified increased with the number of positive nodes. When compared with similar patients, survival curves for those having a single involved node and also those having multiple metastatic nodes were similar whether the node were in the proximal, middle, or distal levels. The 10-year determinate survival for patients with fewer than five positive positive nodes was just under 50%. High positive nodal counts, or involvement of more than one level, were associated with local recurrence of disease.


The influence of untreated internal mammary metastases upon the course of mammary cancer. Internal mammary lymph node biopsies were performed during radical mastectomy in 113 cases of invasive carcinoma of the breast. Metastases were found in 25 cases, 22% of the total. One hundred five of the cases received no further treatment unless cancer recurred and were observed for an average of 13.6 years. Local recurrence and 10-year mortality were significantly increased in the presence of internal mammary metastases, but it could not be demonstrated that the latter increased chest wall recurrence or decreased survival independently of axillary lymph node involvement. The survival of five patients with internal mammary metastases who were treated with postoperative irradiation or castration was not superior to that of 20 patients with untreated metastases. It was concluded that internal mammary metastases usually reflect generalized tumor spread and that local therapy directed at these nodes has a negligible influence upon cure of the disease. In that they reflect a poor prognosis, metastases at this site would justify systemic adjuvant chemotherapy.


Further studies of biochemical predictive tests in breast cancer. This report extends our observations on the relationship between the therapy of breast cancer, estrogen binding and tissue enzymes. We have compared the dextran-coated charcoal method and the sucrose gradient analysis in determining estrogen receptors, and find a sub-group of receptor positive tumors--identifiable only by the latter technique--which we suggest may lead to an incorrect prediction of response to hormonal therapy. Data are also presented to suggest that enzyme analyses show promise in predicting responsiveness to present modes of combination chemotherapy of breast cancer.


Medical and surgical adrenalectomy in patients with advanced breast carcinoma. Twenty-four postmenopausal patients with metastatic breast carcinoma were placed on aminoglutethimide and dexamethasone as a form of reversible medical adrenalectomy. Six patients experienced adverse side-effects. Of the 18 remaining patients 50% had a definite subjective or objective response to therapy. Thirteen of these patients underwent subsequent surgical adrenalectomy after a maximum of 3 month's trial of the medical regimen. In every patient the response to therapy was identical with the two modalities of therapy. In those postmenopausal patients with metastatic breast cancer who are felt to have a hormone- dependent tumor by clinical and/or hormonal assay criteria, medical adrenalectomy may eventually be a feasible replacement for surgery in selected cases. As important, perhaps, is the potential value of this medical adrenalectomy as a reliable indicator of the subsequent response to endocrine ablative therapy.


Lymphatic metastasis with childhood rhabdomyosarcoma. Clinico-pathologic reviews of series of children with rhabdomyosarcoma have yielded conflicting information regarding frequency of lymphatic spread of this disease. The 264 eligible entries in the Intergroup Rhabdomyosarcoma Study (from November 1972 to September 1975) have been categorized by a prospective staging system and pre-treatment characteristics and pathologic findings reviewed. Data accumulated thus far have revealed a higher than expected incidence of lymphatic metastases from extremity (17%) and genito-urinary sites (19%) with a somewhat lower incidence from the orbit (0%), the head and neck region (3%), and trunk (10%). These differences in regard to lymphatic metastases were found to have no relationship to age, sex, tumor size, or histologic type when these factors were simultaneously examined.


Focal nodular hyperplasia of the liver: report of six patients. Concepts of surgical management. The authors add six patients with focal nodular hyperplasia of the liver to the increasing number reported. All had ingested hormones as contraceptives for periods of from one and one-half to seven years. In five patients, the initial manifestation was massive intraabdominal hemorrhage and hypovolemic shock. Five of the six underwent emergency laparotomy for control of hemorrhage. Five patients are living; one patient died six days postoperatively because of coagulopathy. The single diagnostic procedure which proved valuable was liver scan. When hepatic focal nodular hyperplasia is disclosed, we recommend limited resection without ligation of the hepatic artery. Ingestion of birth control hormones should be prohibited.


Endometrial carcinoma in women under 40 years of age: comparison of cases in oral contraceptive users and non-users. Women under 40 years of age rarely develop carcinoma of the endometrium, this age group comprising less than 3% of all cases in most large series. The Registry for Endometrial Carcinoma in Young Women Taking Oral Contraceptive Agents has recorded and studied 30 cases in women under 40 with a history of oral contraceptive administration, and these are compared with 25 cases in the same age group which were accessioned at the Armed Forces Institute of Pathology prior to the era during which these agents have been available, and with series reported in the literature totaling 137 cases with no contraceptive history. In the first group, special attention is paid to the 20 women who received sequential agents exclusively or predominantly, since the association with these agents (primarily Oracon) is much higher than would be expected from the fact that less that 10% of oral contraceptives used in this country are of the sequential type. An analysis of the three groups reveals a similar incidence of the various histologic types (primarily well differentiated tumors with a relatively favorable prognosis), the only difference being the presence of secretory features in 14 of 29 oral contraceptive users vs. only two of 25 women not taking these agents. Clinical and pathologic staging were also predominantly favorable, and comparable in all groups. There were, however, notably higher incidences of nulliparity, obesity, and sclerocystic ovaries--all features traditionally associated with endometrial carcinoma--in the patients not receiving oral contraceptives or receiving combined agents, suggesting that the group receiving sequentials may not represent the same constitutionally predisposed population. Only three of 55 women among our personally reviewed cases have thus far died of cancer (one in the contraceptive group and two in the non-user group) and all three of these had poorly differentiated tumors with myometrial invasion. The excellent prognosis for most patients in this age group is thus confirmed.


Complications following surgery for cancer of the larynx and hypopharynx. The hospital and office records of patients undergoing major surgery for cancer of the larynx and hypopharynx at the Washington University Medical Center, St. Louis from 1955 to 1972 were reviewed. Study parameters were correlated with the incidence of major complications and statistically analyzed to elucidate what factors contribute to increased complication rates. Abnormal margins of surgical resection were found to be significant in determining subsequent complication. Age, sex, race, site and stage of the primary tumor, and the presence of pre-treatment cervical lymph node fixation did not alter the rate of complications. Similarly, low dose preoperative irradiation, various forms of carotid artery protection, and surgical patholgic findings including the size of the tumor, number of positive cervical nodes and cellular characteristics of the tumor showed no significant effect on the rate of complications. Increased total complication rate was associated with an increased death rate. Common complications included wound infection, wound necrosis, salivary fistula, hemorrhage, and carotid artery catastrophe. Also considered were operative deaths and delayed fatal complications. The common causes and treatment of these complications are outlined and safeguards which have been valuable in a sizeable number of patients are discussed.


Squamous cell carcinoma of the tonsil in young adults. Squamous cell carcinoma of the tonsil is rare in people under 40 years of age. Only 11 cases have been treated at the M.D. Anderson Hospital since 1944. Their 5-year survival was decreased (14%) when compared to the total group of patients treated for squamous cell carcinoma of the tonsil (48%). Young adults had higher clinical staging than the total group. Ninety-one percent of the young patients had neck nodal metastasis, with 55% staged N3. Of the entire group, 76% had neck nodal metastasis with 23% staged N3. Case histories indicate a lack of suspicion of tonsillar cancer on the part of patients and physicians. The average total delay from symptoms to diagnosis was 11 months. Response rates of radiotherapy alone were excellent (no failures) in patients with staging T1, T2, TX, N0, and N1. However, five (45%) of the 11 young patients had T3 or T4 primaries and all died, four with regional failure. Six of the 11 young patients had N3 staging and five of these died, all with regional failure except one. Planned combined treatment should be used more frequently in young adults with high staging.


Forty years' evolution in the management of patients with endometrial carcinoma. Of 218 patients with early endometrial cancer treated with planned pre-hysterectomy intracavitary irradiation, 105 were operated 6 weeks after the completion of radiation therapy in the traditional fashion. A group of 113 patients, similar as to stage and differentiation of their disease, were operated on within 48 hours of the removal of the radiation sources. More than 90% of both groups were cured, indicating the lack of benefit from the traditional delayed hysterectomy sequence.


The evolving role of computerized tomography in radiation oncology. The development and application of computerized tomography of the body have important implications in the diagnosis of tumors and in the planning of the radiotherapy treatment of malignancies because the modality provedes: a) an exact contour of transverse sections of the body; b) distinct representation of anatomic internal structures; c) ability to diagnose the presence and extent of tumor involvement of internal organs; d) opportunity to obtain, quantitatively, the "density" (x-ray absorption) of the anatomical structures in the sections; e) capability to develop isodose curves with inhomogeneity corrections; and f) ability to follow the response of a tumor to treatment, non-invasively. Although the diagnostic efficacy of computerized tomography of tumors, relative to other diagnostic modalities, is still to be defined accurately and the exact role of this new method in radiotherapy treatment planning is still to be determined, computerized tomography appears to provide oncologists with a valuable adjunct in their management of tumor patients.


Tumor localization and treatment planning with ultrasound. The successful management of cancer with high energy radiation requires precise knowledge of the tumor position relative to the body surface and normal organs. Diagnostic ultrasound assists in determining the size and location of tumors and vital normal structures in patients undergoing radiation therapy. Radiation fields can be established dynamically under direct ultrasound imaging, and anatomical information about the tumor, normal organs, and body contour utilized in dosimetry calculations. Diagnostic ultrasound is a safe, noninvasive technique of acquiring anatomical information that cannot be obtained by other routine diagnostic methods. Its application in clinical radiation therapy planning has the potential of improving cure rates and decreasing complication rates.


Lymphangiography in the determination of the extent of metastatic carcinoma: the potential value of percutaneous lymph node biopsy. In determining the extent of disease in patients with carcinoma, lymphangiography when read as positive has an accuracy of 90 to 95%. When considered negative 15 to 20% prove to have metastatic disease. the recent use of percutaneous transperitoneal aspiration biopsy of previously opacified lymph nodes has enhanced the value of lymphangiography. Eighty percent of the aspiration biopsies yield sufficient tissue for cytologic diagnosis.


Radiation toxicology: quantitative radiation pathology for predicting effects. Radiation toxicology is the quantification of radiation pathophysiology. It is based upon the development of more precise and accurate radiation dose response data in humans. Such information is particularly important because of the development of combined radiation therapy and chemotherapy. The search for models which will predict radiation injury is presented along with innovative approaches to the 3-dimensional reconstruction of isodose curves in autopsy material, and ultrastructural studies.


The place of irradiation in the management of the primary lesion in head and neck cancers. Lesions of the skin are best excised except for specific locations where surgical excision would not likely produce disease-free margins and/or would require extensive plastic repair. A control rate of 90% for T1 lesions and 80% for T2 lesions is obtained with irradiation in the squamous cell carcinomas of the upper respiratory and digestive tract. Heavy smokers and/or severe alcoholics may be preferably treated surgically. The control rates in the T3 and T4 lesions are unsatisfactory with either irradiation or surgery alone. Pre- or postoperative irradiation combined with the appropriate surgical procedure improves the local control rates and to some extent the survival rates. Postoperative irradiation for the highly malignant tumors of the salivary gland reduces the local failure rate from 36% to 11%. The facial nerve can be preserved unless it is grossly involved.


Treatment of the neck in patients with squamous cell carcinoma of the head and neck. A retrospective study of 702 patients with clinically positive nodes associated with squamous cell carcinoma of the oral cavity, supraglottic larynx, and hypopharynx observed from 1954 to 1968 was done. The policies of treatment for the neck were not standardized during those years. Three hundred eleven patients who survived 24 months with the primary lesion controlled were divided into two groups: 1) those whose neck was treated by surgery alone; and 2) those who had combined radiation therapy and surgery to the neck, to test the efficacy of the two forms of treatment. For the three sites, the recurrence rate in the necks for the surgically treated group was 14% for stage N1, 26% for N2 and 34% N3. Rates for the group receiving combined treatment were 2%, 11% and 25% respectively. Results of the study also showed that elective irradiation, 5,000 rads in five weeks, will prevent metastasis from occurring in the NO staged neck.


Radiation therapy of brain tumors.

Radiation therapy of brain tumors. Results of radiation therapy obtained at the University of California, San Francisco, over approximately the past 20 years for various histologic types of brain tumors are presented. Included are astrocytomas, malignant gliomas, medulloblastomas, ependymomas, oligodendrogliomas, and brain stem tumors. Degree of malignancy and tendency to disseminate within the central nervous system are also reviewed. For each tumor type and grade considered, the survival rate appeared improved when incomplete resection was followed by irradiation. The increase in survival rate for glioblastomas was only evident for 1-2 years, but for the remainder the improvement extends to 5-10 years. Since many patients were still alive at the time of review, it is possible that permanent control of many intracranial neoplasms may be induced by radiation therapy.


Surgical considerations in colorectal cancer. The large bowel, functioning largely as a wast storage receptacle, and with its long mesentery permits wide local excision and liberal removal of the lymphatics and blood vessels following the principles of cancer surgery. Rectal lesions with no mesentery or peritoneum, located in the narrowing lower pelvic cone are exceptions. With improved general patient support, antibiotics and technical improvements, 5-year survival in patients with localized disease now approaches 70%. "No-touch" principles with early ligation of the vascular pedicle, constricting tapes above and below the lesion, and local management of the resected bowel margins, have presumably diminished the dissemination of tumor cells at operation and lessened the chance of local implantation. Abdominoperineal resection, first described by Miles in 1908, is being reassessed because operative mortality and morbidity with a permanent colostomy is high, especially in the elderly patient. Local treatment by electrofulgeration, cryosurgery or radiotherapy are under intensive study as possible alternatives to surgery. The overall national death rate has improved only slightly during recent years. Thus the search for effective systemic measures to destroy small foci of cancer cells beyond the surgeon's reach must continue at a vigorous pace.


Tumorigenic agents in unburned processed tobacco: N-nitrosodiethanolamine and 1,1-dimethylhydrazine. Two tumorigenic agents, N-nitrosodiethanolamine and 1,1-dimethylhydrazine, have been isolated from tobacco for the first time. The former, a reportedly weak hepatic carcinogen in rats, varied in amounts from a low of 0.1 ppb in flue-cured tobacco not treated with the herbicide MH-30, to a high of 173 ppb in Burley tobacco to which the herbicide had been applied prior to harvesting. MH-30 (maleic hydrazide) used by farmers to remove 'suckers' from tobacco plants, is commonly formulated as the diethanolamine salt. 1,1-Dimethylhydrazine, reported to induce tumors in mice, ranged in amounts from 60 to 147 ppb, except in the case of Burley tobacco where none was detected (detection limit: 0.1 ng). The source of the nitrosamine in the tobacco appears to be the MH-30, whereas that of dimethylhydrazine has not been determined.


Inhibitory effects of selenium on 1,2-dimethylhydrazine and methylazoxymethanol acetate induction of colon tumors. Sprague-Dawley rats were injected weekly with either 1,2-dimethylhydrazine (DMH) or methylazoxymethanol acetate (MAM). Addition of 4 ppm selenium (sodium selenite) in the drinking water reduced the number of rats developing DMH-induced colon tumors from 13 to 6 groups of 15 each. The total number of tumors observed in these two groups was 39 in the DMH treated and 11 in the DMH plus selenium. The incidence of MAM-induced tumors was 93% (14/15) with the selenium additive and 100% (14/14) when MAM was administered without the selenium supplement. However, selenium decreased the total number of colon tumors induced by MAM to 42 tumors as compared to a total of 73 tumors in rats receiving only MAM. Both carcinogens induced tumors with a higher frequency in the transverse colon as compared to either the proximal or distal colon. Selenium at this level did not affect the weight gain of the animals.


High multiplicity of H1 histones from mouse mammary tumors. Lysine-rich (H1) histones were isolated from mammary tumors of GR mice. Chromatography of the H1 histone class on a long column (130 X 1 cm) of Amberlite IRC-50 resin revealed the presence of nine components, designated 1-9 according to their order of elution. Amino acid analysis indicated that components 2-9 represented H1 class histones, but that peak 1 represented a nonhistone protein. Studies of three types of mouse mammary tumors, viz. hormone-dependent, hormone-responsive and hormone-independent, did not reveal significant differences in elution profiles of the H1 histones. In contrast, the H1 histone class from a dog mammary tumor differed chromatographically from that of mouse mammary tumors, indicating species specificity in elution patterns of mammary tumor H1 histones.


Induction of aryl hydrocarbon hydroxylase in human pulmonary alveolar macrophages and peripheral lymphocytes by cigarette tars. Cigarette smoking is associated with alterations in pulmonary alveolar macrophages (PAMs), including increased cytoplasmic inclusions and induction of the aryl hydrocarbon hydroxylase (AHH) system. Nonpigmented PAMs from nonsmokers were able to ingest and accumulate pigment from lysed PAMs of smokers, however, this pigment did not induce AHH activity in either PAMs or peripheral lymphocytes. In contrast, the cigarette tars significantly induced AHH levels in PAMs and in peripheral lymphocytes from either nonsmokers or smokers. This provides further evidence that components in cigarette smoke can explain the in vivo induction of AHH documented in cells from smokers.


A study of tobacco carcinogenesis. XV. Effects of N'-nitrosonornicotine and N'-nitrosoanabasine in Syrian golden hamsters. N'-Nitrosonornicotine (NNN) and N'-nitrosoanabasine (NAB) were synthesized and injected subcutaneously three times weekly in male and female Syrian golden hamsters for 25 weeks (total dose 375 mg). N-Nitrosopiperidine (NPD) served as positive control (total dose 150 mg). Within 83 weeks 12 out of 19 hamsters given NNN had developed tracheal tumors, 1 had a carcinoma of the nasal cavity. During the same time none of the hamsters given NAB developed tumors, whereas all 20 hamsters given NPD developed tracheal tumors and about 50% had additional neoplasms in the nasal cavity. Under the described conditions and in comparison to NPD, NNN appears to be a moderately active tumorigenic agent in the upper respiratory tract of Syrian hamsters, whereas NAB is inactive.


Evaluation of estrogen receptor assays in human breast cancer tissue. Standard dextran-coated charcoal (DCC) and sucrose gradient centrifugation assays for estrogen receptor were compared in a series of human breast cancer tissues. From a quantitative standpoint the results were remarkably similar. A simplified version of the DCC assay compared to the sucrose gradient assay yielded acceptable results. We conclude that, in spite of the lack of specificity controls inherent in the sophisticated standard assays, the simplified DCC assay might be useful if the biopsy specimen is too small to provide the number of aliquots for a standard DCC assay or sufficient protein for a sucrose gradient analysis. It also might be useful in research laboratories attempting to develop assays for multiple receptors or other constituents in a single tumor biopsy specimen.


Macromolecular binding of glucocorticoids in human mammary carcinoma. The presence of glucocorticoid receptors was examined in 100 primary and 22 metastatic human breast cancer lesions. Aliquots of cytosol were incubated in vitro with tritiated cortisol and dexamethasone with and without competing steroids. Two different types of glucocorticoid receptors were detected. One is similar to transcortin; it sediments at 4 S in the ultracentrifuge, has a dissociation constant in the same range (10(-8) M), and does not bind fluorinated corticosteroids. While every tumor showed cortisol binding, very high activity (greater than 1000 fmoles/g tissue) was detected in 38% of primaries and in 59% of metastases. Plasma transcortin could be excluded as the source of binding activity. The other receptor binds both natural and fluorinated corticosteroids, has a higher affinity (Kd 10(-9) M), and sediments at 8 S. It was present in 23% of tumors and its quantity (26 to 995 fmoles/g) was much less than that of cortisol binder (50 to 6000 fmoles/g). While there was no correlation between the two glucocorticoid receptors, 80% of dexamethasone receptor-positive cases also had estrogen receptor. The results indicate that a significant proportion of these tumors could be subject to glucocorticoid influence.


Free DNA in the serum of cancer patients and the effect of therapy. A radioimmunoassay for ng quantities of DNA was developed. [125l]lododeoxyuridine-labeled DNA was used as the antigen, and the serum of a lupus erythematosus patient served as the source of antibody. The level of free DNA in the serum of 173 patients with various types of cancer and in 55 healthy individuals was determined by this radioimmunoassay. DNA concentration in the normal controls had a range of 0 to 100 ng/ml with a mean of 13 +/- 3 ng/ml (S.E.). For comparison purposes, the range of 0 to 50 ng/ml was designated as normal, and 93% of controls were found in this range. In the cancer patients, the DNA concentration ranged from zero to mug levels with a mean of 180 +/- 38 ng/ml. Fifty % of the patients values were found in the range of 0 to 50 ng/ml; the other 50% were between 50 and 5000 ng/ml. No correlation could be seen between DNA levels and the size or location of the primary tumor. Significantly higher DNA levels, however, were found in the serum of patients with metastatic disease (mean of 209 +/- 39 ng/ml), as compared to nonmetastatic patients (mean 100 +/- 30, p less than 0.02). After radiation therapy in lymphoma, lung, ovary, uterus, and cervical tumors, the levels decreased in 66 to 90% of the patients, whereas in glioma, breast, colon, and rectal tumors, the DNA levels decreased only in 16 to 33% of the patients. Generally, the decrease in DNA concene of tumor size and reduction of pain. Conversely, when DNA levels either increased or remained unchanged, a lack of response to the treatment was noted. Of 17 patients who died within a year, 13 showed DNA levels that remained high or unchanged, whereas only 4 showed lower levels during treatment. Persistent high or increasing DNA levels in the circulation, therefore, may signal a relapse and are probably a poor prognostic sign. The relatively high percentage (50%) of cancer patients with apparently normal DNA levels would suggest that this test may have low diagnostic value. It should be pointed out, however, that all these patients represent a selected group considered for radiation therapy, usually after surgery and/or chemotherapy. It is possible that a better correlation between DNA levels and cancer will be obtained prior to the initiation of treatment. On the other hand, DNA in the serum may be an important tool for the evaluation of therapy or the comparison of different regimens.


Increased susceptibility of cytosol proteins to proteolytic digestion during regression of a hormone-dependent mammary tumor. Regression of MTW9 mammary carcinoma, which consistently follows withdrawal of mammotropic hormones, was characterized by a rapid decrease of thymidine incorporation into DNA but only a slight reduction or uridine incorporation into RNA and amino acid incorporation into proteins. Within 24 hr of hormone withdrawal, cytosol proteins of MTW9 became more easily degraded by trypsin, alpha-chymotrypsin, or subtilisin BPN'. Labilization of cytosol proteins occurred much earlier than any change in the level of protein synthesis or lysosomal enzyme activity. The data showing increased susceptibility to proteolysis could not be explained either by the presence of endogenous proteases, by the destruction of the exogenous proteases used in the assay, or by the existence of protease inhibitors. Nor were any differences detected either in the distribution of radioactive precursor among the cytosol proteins from growing or regressing tumors or in the electrophoretic pattern of the same proteins. Preincubation of the cytosol proteins with dithiothreitol or with prolactin, 17 beta-estradiol, progesterone, and hydrocortisone did not modify the susceptibility to proteolysis. However, after heat denaturation, cytosol proteins of regressing and growing tumors became equally susceptible to proteolysis. It is suggested that regression of MTW9 mammary carcinoma occurs not only because cell reproduction is arrested, but also because susceptibility of cytosol proteins to proteolysis is increased.