Adrenaline levels in brain stem nuclei and effects of a PNMT inhibitor on spontaneously hypertensive rats. The adrenaline content of specific brain stem areas was decreased in young (4 week-old) but not in adult (14 week-old) SHR. Some of these areas also showed increased PNMT activity, indicating a possible enhanced release or metabolism of adrenaline early in the development of the hypertension. Dopamine and noradrenaline levels, on the contrary, were not changed either in young or adult animals. Administration of a PNMT inhibitor to adult SHR resulted in a decrease of the blood pressure in SHR to control levels. These observations support the hypothesis of a participation of adrenergic mechanisms in the development of genetic hypertension, and indicate the possibility for the use of PNMT inhibitors as hypotensive agents.


Electrophysiological and biochemical responses of noradrenergic neurons to a non-amphetamine CNS stimulant. Amfonelic acid (AFA), a potent non-amphetamine CNS stimulant, has been shown previously to have marked effects on dopamine (DA) metabolism and DA neuronal activity, but no effect on norepinephrine (NE) metabolism. AFA is known to inhibit the NE neuronal uptake mechanism. Other NE uptake inhibitors, such as desipramine (DMI), have been shown to decrease the firing rate of NE-containing locus coeruleus (LC) neurons. The purpose of the present study was to compare the actions of AFA and DMI electrophysiologically on LC neurons, and biochemically on NE metabolism in whg rate, with DMI being more potent. Brain NE metabolism was not influenced by either AFA or DMI at doses considerably higher than those  which were effective in reducing NE neuronal impulse flow. Thus, NE uptake inhibition coupled with a decrease in impulse flow results in no net change in NE metabolite formation. The effects of AFA on LC unit activity do not seem to be due to its marked effects on brain DA, since DA receptor blockade with haloperidol had little effect on LC unit responsiveness to AFA (or amphetamine). Whereas AFA has dramatic effects on DA metabolism via enhanced release per impulse, the drug has minimal effects on NE metabolism, and this specificity of action may be related to differences in NE and DA transmitter storage mechanisms. It is concluded that the effects of AFA on NE neuronal firing rate are likely due to the drug's DMI-like action and not to enhanced NE release per impulse.


The dorsal bundle extinction effect:dependence on subtle changes in acquisition. Destruction of the ascending noradrenergic innervation to the forebrain in rats by intracerebral injection of the selective neurotoxin 6-hydroxydopamine (4 microgram in 2 microliter injected bilaterally into the dorsal bundle in the mesencephalon) was found to cause resistance to extinction of a continuously reinforced lever press response. However, this effect occurred only if the lesion were present during acquisition training on the reinforced schedule and not if intact animals were trained and the lesion inflicted after completion of acquisition training and just prior to the extinction phase. Thus, the behavioural effect that manifests itself during extinction appears to be due to subtle changes in the acquisition learning process. This is consistent with the predictions of an attentional theory of noradrenergic function and appears to exclude most other suggested explanations of the dorsal bundle extinction effect.


Characterization of proteins associated with nuclear ribonucleoprotein particles by two-dimensional polyacrylamide gel electrophoresis. Rat liver nuclear ribonucleoprotein particles were prepared by two different methods and defined as 40S ribonucleoprotein (40S RNP) and heterogeneous nuclear ribonucleoprotein (HnRNP) particles. The RNP particles were either solubilized in 8 M urea--6 mM 2-mercaptoethanol--20 mM glycine--20 mM Tris--HCl (pH 8.4) or subjected to removal of RNA by phenol extraction prior to solubilizing the proteins in the urea buffer. The proteins associated with 40S RNP and HnRNP were heterogeneous and very similar in their electrophoretic patterns when analyzed by two-dimensional PAGE, except a protein with molecular weight of 62 000 and an isoelectric point (pI) of 6.2 was present only in HnRNP particles. At least 12 major and 22 minor components could be identified in both preparations. The major proteins were found at pI values varying from 6.0 to 8.5 and with molecular weights from 32 000 to 42 000, and a group of proteins with molecular weight approximately 65 000 were more prominent in HnRNP than in 40S RNP. The other components were found mainly at pI ranges from 5.0 to 6.5 with molecular weights from 43 000 to 65 000. The phenol method extracted essentially all proteins associated with either 40S RNP and HnRNP, but was less effective in extracting a group of proteins with pI values from 5.0 to 5.5 and more efficient for proteins with pI values from 7.5 to 8.5. When chromatin proteins isolated by phenol extraction were compared with HnRNP particle proteins isolated by the same method, the electrophoretic mobilities of the HnRNP particle proteins were found to be identical with a fraction nonhistone chromatin proteins. The 40S RNP particles were further purified by metrizamide isopycnic density gradient centrifugation. The electrophoretic patterns of these proteins were very similar to those prepared by sucrose density gradient centrifugation. Therefore, we concluded that the proteins of RNP particles constituted part of the chromatin proteins.


Histochemical and cytochemical localization of acetylcholinesterase in retina and optic tectum of teleost fish. The activity of cholinesterase and its cellular and subcellular localization were investigated in the retina and optic tectum of Eugerres plumieri and in the retina of Carassius carassius by means of radiometric, histochemical, and cytochemical procedures. In both fishes only the presence of acetylcholinesterase could be demonstrated. This study, besides confirming previous findings that acetylcholinesterase is located in the ganglion and amacrine cells of the retina as well as in the inner plexiform layer, in addition provides evidence that the enzyme is also present at the region of photoreceptor synapses between the cell bodies and apposing extensions of the horizontal cells of the same layer. The latter localization may indicate the involvement of a cholinergic mechanism at the functional contacts (transferapses) between the horizontal cells. In the optic tectum of Eugerres plumieri, histochemistry reveals fine distinguishable bands of acetylcholinesterase activity; two of the bands are quite sharply defined, whereas three others have rather a more diffuse appearance. The presence of these bands and their distribution may suggest a widespread distribution of cholinergic elements in the optic tectum.


Effect of phalloidin on actin proteolysis as measured by viscometry and fluorimetry. Even at low concentration, phalloidin shows a marked protection of F-actin against the action of trypsin or pronase. G-actin is not protected at any concentration of phallodin. The kinetics of the proteolysis show that a change in the environment of tryptophan residues is preceded by disruption of the filamentous structure of F-actin.


Relationship between tissue levels of S-adenosylmethionine, S-adenylhomocysteine, and transmethylation reactions. The concentrations of S-adenosylmethionine (AdoMet), S-adenosylhomocysteine (AdoHcy), and various methyltransferases were determined in the cerebrum, cerebellum, and liver of rats during development and aging. The liver contained from 3 to 7 and from 10 to 15 nmol AdoHcy per gram in young and adult rats, respectively. The AdoMet concentration was 60 to 90 nmol/g liver from rats of the same age and sex. It did not vary significantly with age. In the brain the AdoMet concentration was 45 to 50 nmol/g at birth and decreased to 20 nmol/ g tissue with maturity of the organ. The level of AdoHcy in this organ was less than 1 nmol/g tissue throughout the life-span of the rat. Since the ratio of AdoMet to AdoHcy is relatively high, the rate of methylation of histones, DNA, or phosphatidylethanolamine in the liver or brain was not significantly influenced by AdoHcy. Under normal nutritional conditions, the tissue concentration of AdoMet is far above the Km values of histone and phosphatidylethanolamine methyltransferases. The levels of activity of these enzymes in liver and brain did not correlated with the cellular concentration of AdoHcy. Thi histone methyltransferase activity was elevated in rapidly proliferating tissues and declined markedly in the absence of histone biosynthesis. Phosphatidylethanolamine methyltransferase activity was elevated during development of the liver. The specific activity of the AdoHcy hydrolase remained relatively constant in the rat brain and liver. The activity of this enzyme was 10 times higher in liver than in brain, yet the concentration of AdoHcy was much lower in the latter organ. The tissue levels of this compound are evidently dependent on the rates of removal of homocysteine and adenosine. Adenosine deaminase was present in the liver and brain at relatively high concentrations, particularly during development.


Effect of lysolecithin in solubilizing membrane-bound glycosyltransferases. Microsomal membranes were solubilized by incubation with lysolecithin which caused considerable release of galactosyl- and N-acetylglucosaminyl-transferase into a high-speed supernatant fraction. With a critical concentration of lysolecithin (2.5 mg/10 mg protein in 1 mL microsome suspension), there was a maximal binding of radioactive lysolecithin to the sediment fraction obtained after high-speed centrifugation. Increase of lysolecithin concentration (above 2.5 mg/mL) in the incubation mixture caused a progressive release of the enzymes into the supernatant fraction. Lysolecithin binding to the membrane was greatly inhibited by 1 M NaCl, and high salt concentration also inactivated galactosyltransferase in the sediment, suggesting an electrostatic interaction between lysolecithin and membrane enzyme. In contrast, high NaCl concentration had no inhibitory effect on the enzyme activity in the sediment when the fraction was prepared by treatment with Triton X-100. Lysolecithin-treated microsomal sediment and supernatant galactosyltransferase was inactivated by oleoyllysophosphatidic acid but not by palmitoyllysophosphatidic acid or egg yold lysophosphatidic acid. Triton X-100 treated microsomal fractions were also similarly affected by different species of lysophosphatidic acid. The results suggested a similarity of interactions of lysophosphatidic fatty acyl species with lysolecithin and Triton-treated galactosyltransferase.


A structural basis for four distinct elution profiles on concanavalin A--Sepharose affinity chromatography of glycopeptides. Twelve 14C-acetylated glycopeptides have been subjected to affinity chromatography on concanvalin A (Con A)--Sepharose at pH 7.5. The elution profiles could be classified into four distinct patterns. The first pattern showed no retardation of glycopeptide on the column and was elicited with a glycopeptide having three peripheral oligosaccharide chains: (abstract:see text). Such glycopeptides have only a single mannose residue capable of interacting with Con A--Sepharose; an interacting mannose residue is either an alpha-linked nonreducing terminal residue or an alpha-linked 2-O-substituted residue. The second type of profile showed a retarded elution of glycopeptide with buffer lacking methyl alpha-D-glucopyranoside (indicative of weak interaction with the column) and was given by glycopeptides with the structures: (abstract: see text) where R1 is either H or a sialyl residue. The third profile type showed tight binding of glycopeptide to Con A--Sepharose and elution as a sharp peak with 0.1 M methyl alpha-D-glucopyranoside; glycopeptides giving this pattern had the structures: (abstract: see text) where R2 is either H, glcNAc, Gal-beta 1,4-GlcNAc, or sialyl-Gal-beta 1,4-GlcNAc. These glycopeptides all have two interacting mannose residues, the mimimum required for binding to the column; one of these mannose residues must, however, be a terminal residue to obtain tight binding and sharp elution. The fourth profile type showed tight binding of glycopeptide to the column but elution with 0.1 M methyl alpha-D-glucopyranoside resulted in a broad peak indicating very tight binding; glycopeptides showing this behaviour had the structures: (abstract: see text) where R3 is either GlcNAc,Gal-beta 1,4-GlcNAc, or sialyl-Gal-beta 1,4-GlcNAc. Therefore it can be concluded that although a minimum of two interacting mannose residues is required for binding to Con A--Sepharose, the residues linked to these mannoses can either strengthen or weaken binding to the column.


Heterogeneous catalysis by phospholipase A2: formulation of a kinetic description of surface effects. The kinetics of hydrolysis of aqueous dispersion of long-chain, saturated phosphatidylcholines (PC) catalysed by Crotalus atrox phospholipase A2 (PLA) have been analyzed, and a reaction mechanism proposed which takes surface effects into account. PLA is proposed to form an enzyme-substrate complex with surface substrate molecules, thereby undergoing a conformational change which exposes sites that interact with the lipid surface. After a hydrolytic event, the enzyme can either desorb from the surface (path 1), or diffuse along the surface to an adjacent substrate molecule (path 2). The path 1 dominated mechanism leads to Michaelis-Menton steady-state kinetics, and characterizes hydrolysis of gel phase PC. Evidence for saturation of the surface with PLA was obtained at high enzyme concentrations. The path 2 mechanism dominates when the desorption rate is very small; this mechanism describes hydrolysis of liquid crystalline phase PC and is characterized by an initial burst of hydrolysis followed by a very slow reaction. The velocities in these two phases of the reaction are independent of bulk PC concentration. When gel and liquid crystalline PC phases coexist, as in mixtures of dimyristoyl- and distearoyl-PC, the liquid crystalline phase is preferentially hydrolysed. Products of the reaction (lyso-PC and fatty acid) stimulated hydrolysis, apparently by stimulating desorption of PLA. The desorption rate constant appears to be a linear function of the surface concentrations of lyso-PC and fatty acid. The proposed model describes hydrolysis progress curves extremely well and is consistent with current ideas on the mechanism of catalysis by this enzyme.


Experimental oral and nasal transmission of rabies virus in mice. Weanling female white Swiss mice were exposed to challenge virus standard rabies virus and street virus isolates from various domestic and wild animals. Virus was given free choice as suspension or as infected mouse brain by stomach tube, by single injection of suspension into the oral cavity of unanesthetized mice, by repeated injection into the oral cavity of anesthetized mice and by single application to the external nares of anesthetized mice. Challenge virus standard virus in mouse brain suspension and a suspension of skunk salivary glands infected with street virus (titers greater than or equal to 10(6)MICLD50/0.03 ml) consistently produced high rates of infection in mice exposed intranasally, low to high rates of infection in mice exposed by forced feeding and other artificial methods of oral exposure and very low rates of infection when given free choice. Street virus isolates passaged intracerebrally in mice had titers less than or equal to 10(4.5) MICLD50/0.03 ml and rarely caused rabies in mice exposed orally or nasally by any method. The results indicate that with the isolates used, virus of high titer (greater than or equal to 10(6)MICLD50/0.03 ml) is required to consistently produce infection in mice by the nasal route and that the mucosa of the nasal cavity probably is the chief route of infection even after oral administration.


Anastomoses of intestinal tract as a sequela to rectal stricture in swine. Rectal stricture with anastomoses of colon and/or small intestine to a spherical dilatation of the descending colon proximal to the stricture was observed in six swine. In three of these swine, feces bypassed most of the colon in passage from the small intestine to the dilated colon. Rectal stricture with an anastomosis of a loop of colon to the rectal stricture was observed in one pig where feces bypassed much of the stricture.


Prenatal infection following maternal exposure to porcine parvovirus on either the seventh or fourteenth day of gestation. Intranasal and oral exposure of two gilts to porcine parvovirus on either the seventh or 14th day of gestation resulted in prenatal infection. Normal appearing fetuses and necrotic remnants of what were believed embryos and extraembryonic membranes were found when the gilts were necropsied seven weeks after exposure. The presence of masses of porcine parvovirus antigen throughout necrotic tissues of six of seven embryos, but not in any of the nine normal appearing fetuses suggested that embryonic death was due to porcine parvovirus.


Structural abnormalities of the X-chromosome in a heifer. Structural chromosome abnormalities, some of them unusual, were found in 18 (36%) of 50 photomicrographed mitotic cells of a blood lymphocyte culture from a three month old Holstein heifer affected with chronic tympanism of the rumen. The aberrations involved more specifically the X-chromosome. The etiology of the anomalies and the cause of their X-chromosome propension are of unknown nature.


Brain viral persistence and myelin damage in nude mice. Multiple foci of secondary demyelination were observed in the cerebellum of nude mice given two inoculations of avirulent Semliki Forest virus compared to single lesions seen in similarly treated Swiss A2G mice. The increased number of demyelinative plaques were attributed to brain viral persistence for 35 days in the nude mice with correspondingly low serum antibody titres. No neurological signs were observed in any of the mice.


The effect of vasoactive intestinal polypeptide on the urinary bladder and taenia coli of the guinea pig. The effect of vasoactive intestinal polypeptide (VIP) on the contractile activity of the urinary bladder and taenia coli of the guinea pig has been investigated. In the urinary bladder VIP caused a weak contraction and a small potentiation of the nerve-induced contraction. In the taenia coli VIP caused a small relaxation and had no effect on the nerve-induced relaxation of the preparation. The present experiments provide no evidence that VIP fulfills a role as a neurotransmitter in the urinary bladder or taenia coli, but it could be a modulator of transmitter action in the urinary bladder.


Human lymphocyte migration in vitro: characterization and quantitation of locomotory parameters. Using a time-lapse cinephotomicrographic technique, the locomotory paths taken by lymphocytes have been shown to satisfy the requirements for a continuous-time Markov chain analysis consisting of four directional states defined by the four quadrants of a Cartesian plane and state 0 in which the cells are stationary. The important parameters characterizing this path motion are the average time (waiting time) that a cell spends in each state and the state transition probabilites which, using the Markov model, allow the computation of the probability of the cells moving in a given direction. Our results show that lymphocytes moving alone on glass exhibit a random migration pattern.


Voltage control during inward current flow in rat ventricular muscle using a double sucrose gap technique. In rat ventricular muscle, measurements of the membrane potential with microelectrodes during depolarizing voltage steps showed that deviation of the membrane potential from the command signal were never larger than 15 mV during flow of the fast inward current and that voltage control was regained within 15 ms after the beginning of the voltage step. During the flow of the slow inward current, tail currents elicited by interrupting the time course of the slow current at different time intervals returned exponentially to the steady-state level, thus indicating acceptable voltage control. It is concluded that rat ventricular muscle is a rather favorable preparation for voltage-clapm experiments and this is attributed mainly to the geometry of the preparation.


Effects of penicillin pretreatment on renal tubular para-aminohippurate transport in the immature rat. The effect of pretreatment with penicillin on para-aminohippurate (PAH) transport by the kidney of the immature rat was evaluated in vivo. After 3 days of penicillin administration, renal clearances of inulin (CIN), PAH (CPAH), and the renal tubular transport maximum (Tm) for PAH were measured in rats as young as 17 days of age. The CPAH in 19- to 21-day-old rats pretreated with procaine penicillin was 54% greater than that of their littermate controls. Similarly, CPAH of rats that received sodium penicillin was 31% greater than control. CIN was not increased after penicillin pretreatment. Pretreatment of rats older than 24 days did not change CIN or CPAH. The Tm for PAH of 17-day-old rats pretreated with sodium penicillin was 51% greater than that of control rats. It was concluded that pretreatment with penicillin enhances the renal secretion of organic anions by the immature rat.


Developmental abnormalities induced by 6-mercaptopurine in the hamster. Pregnant hamsters were given varying doses of 6-mercaptopurine (6MP) at various times during gestation. The fetuses were examined for both gross and histological malformations which showed that the toxic and teratogenic effects of 6MP were dose and time dependent. The most severe gross malformations induced by 6MP were cleft palate, micrognathia and agnathia, microglossia, short limbs, and gut herniation. Grossly normal appearing fetuses, greated during late gestation, showed malformations at the tissue and cellular level. The effects of 6MP in hamster was compared with other species, and with other growth-supressive agents, and it was deduced that the teratogenicity of 6MP is species and tissue specific. Also, it was recommended that histological observations be made an integral part of the teratological safety analysis.


Increased pressor responses to pressor agents in spontaneously hypertensive rats. Pressor reactivity to a variety of pressor agents after partial ganglionic blockade induced with hexamethonium was investigated in intact, in spinalized, and in chemically sympathectomized, spontaneously hypertensive rats (SHR). Responses of unanaesthetized 6-month-old SHR to noradrenaline, phenylephrine, and angiotensin after hexamethonium administration (32 mg/kg) markedly exceeded those of unanaesthetized, age-matched normotensive Wistar-Kyoto rats (WKR). Responses of anaesthetized SHR to noradrenaline after hexamethonium administration (16 mg/kg) were also increased at the hypertensive stages but not at the prehypertensive stages, when compared with those of anaesthetized normotensive Wistar rats of respective ages. In spinalized and chemically sympathectomized preparations after hexamethonium administration (16 mg/kg), noradrenaline produced equal increases in blood pressure in 6-month-old SHR and WKR. It is suggested that the functional sympathetic nervous system is important for the hyperreactivity of intact SHR.


Cardiac edema in dogs: distribution of renal blood flow and glomerular filtrate. The injection of Freund's adjuvant into the pericardial sac of 29 dogs resulted in chronic pericardial tamponade with persistent sodium retention. Micropuncture, clearance, and radioactive microsphere experiments were initiated 6--13 days after pericardial injection and 60 min after pericardiocentesis. Pericardiocentesis increased sodium excretion (from 12.2 to 41.3 microequiv./min) and mean arterial pressure (+ 20 mmHg (1 mmHg = 133.322 Pa)). Central venous pressure decreased 6.5 mmHg, as did hematocrit (from 45.7 to 39.8%) and plasma protein concentration (from 5.88 to 5.15 g%). Pericardiocentesis had no significant effect on renal blood flow (RBF), nor plasma flow. Redistribution of glomerular filtrate was suggested by the observation that superficial nephron glomerular filtration rate increased (from 91 to 108 nL/min) while glomerular filtration rate remained unaltered. Determination of intrarenal distribution of RBF revealed that cortical blood flow also distributed superficially. A significant increase in the fraction of RBF perfusing zone 1 (outer cortex) and a decrease in fractional perfusion of zones 2, 3 and 4 (juxtamedullary cortex) were observed in each experiment following pericardiocentesis. RBF distribution examined in a series of six animals prior to and during the development of pericardial tamponade showed the opposite effect. These results indicate that pericardiocentesis causes redistribution of both glomerular filtrate and RBF to superficial nephrons. The development of pericardial tamponade was associated with increased fractional juxtamedullary blood flow. These changes may have been the result of altered blood pressure, hematocrit, plasma protein concentration, or altered renal resistance.


Cardiovascular actions of kinins in the rabbit. The hypotensive action of bradykinin (BK) and congeners was measured in anesthetized rabbits by administering the peptides intravenously and intraarterially in order to evaluate their pulmonary inactivation. A systematic study of the distribution of receptors for BK in the cardiovascular system of the rabbit was approached: (a) by measuring the myotropic effects of several peptides related to BK in strips of large arteries and veins; (b) by recording the changes of tension and rate of isolated atria; and (c) by evaluating the changes of perfusion pressure in isolated hearts, kidneys, and ears. This investigation was extended to strips of aortae of various mammals and to isolated atria of guinea pigs, for comparison. Receptors for BK were classified into two main types, B1 and B2, using the order of potency of these agonists [Tyr(Me)8]-BK, BK, and [des-Arg9]-BK, and an antagonist, specific and competitive for the B1 receptors, the octapeptide [Leu-OMe8,des-Arg9]-BK. The results obtained in this study indicate that the complex cardiovascular effect of BK in vivo may result from direct actions on vascular smooth muscles, presumably mediated by at least two types of receptors, as well as from the release of endogenous prostaglandins. BK and congeners exert a direct action on vascular smooth muscle by stimulating specific receptors both of the B1 type (in the aorta, the large arteries, and the mesenteric vein) and of the B2 type (in the jugular vein); and these vascular tissues provide useful preparations for pharmacological studies of bradykinins. Isolated organs perfused through their main arteries with physiological medium respond to BK by an increase of perfusion pressure (vasoconstriction in isolated ears and kidneys) or by a decrease (vasodilation in the rabbit heart). The vascular effects of BK in the heart and the kidney depend in part on the release of endogenous prostaglandins and on the activation of receptors that appear to be of the B2 type. Like other endogenous hypotensive agents, BK appears to reduce the tonus of the peripheral vessels, while contracting large arteries and veins. The results obtained in vitro are discussed with respect to the hypotensive effect in vivo and to the role of kinins in inflammation and oedema.


Phophate-induced renal calcification in the rat. Studies were done to investigate nephrocalcinosis produced in weanling female Wistar rats fed pelleted, semisynthetic diets. The rats were fed diets varying in concentrations of Ca and P supplied as inorganic salts for periods of 4--6 weeks and results compared with control rats fed laboratory rodent chow for the same period of time. Measurement of renal Ca and P concentrations showed that nephrocalcinosis was produced by semisynthetic diets with inorganic phosphate concentrations as low as 0.5% on a weight basis; in contrast, rats fed regular laboratory chow (P = 0.72%) showed no evidence of nephrocalcinosis. The severity of the lesion was proportional to dietary phosphate concentrations from 0.5 to 1.0% but other dietary factors modified the severity of the lesion. With the lower dietary phosphate of 0.5%, increasing dietary Ca from 0.5 to 1.0% decreased the severity of the renal calcification. Decreasing protein concentrations from 25 to 15% casein increased the severity of the renal lesions. Other dietary factors also appear to modify the phosphate-induced nephrocalcinosis since no lesions occurred in rats on laboratory chow. It is suggested that the availability of dietary phosphate may be a factor. The phosphate in the semisynthetic diets was totally inorganic while the natural foods of laboratory chow contain, at least in part, organic phosphate.


Superiority of a soil debris isolation method over a beet seed colonization method for assay of Rhizoctonia solani at high soil inoculum densities. A quantitative soil debris isolation method (all debris from known weight of soil plated) and a garden beet seed saprophytic colonization method were compared over a 1-year period for assaying Rhizoctonia solani population. Four fields of different soil textures were selected. Within each field four areas of healthy and four areas of diseases (rhizoctonia root and crown rot) sugarbeets were sampled bimonthly from August 1976 until June 1977. The maximum numbers of R. solani colonies obtained by the debris method were 2 per gram of soil in areas of healthy beets, and 11 per gram of soil in areas of diseased sugarbeets. At such high inoculum densities the beet seed colonization method underestimated R. solani populations, because the inoculum per unit of soil exceeded the numbers of beet seeds per unit of soil available for colonization. Modifications of the beet seed method did not significantly alter results of colonization assays. Ranked correlation comparisons of assay methods yielded r = 0.81 for all data.


Bacteriophage-like particles associated with a spirochete. A lysed Treponema bacterium containing cubic phage-like particles, approximately 40nm in diameter, has been observed by negative staining electron microscopy.


The physiological effects of restrictive environmental conditions on Dictyostelium discoideum spore germination. Spores may be reversibly activated by the application of heat, dimethyl sulfoxide, urea, or ethylene glucol. Severe changes in four environmental variables (high osmotic pressure, low oxygen tension, low or high pH, and low or high temperature) interfere with the germination process. Spores at the end of the postactivation lag phase of germination were usually deactivated if exposed to severe environmental conditions and thus did not swell; spores in the swelling and oxygen uptake which began during spore activation was primarily attributable to a cyanide-sensitive pathway and secondarily to a salicylhydroxamic acid (SHAM) sensitive pathway. Inhibition of the SHAM-sensitive pathway did not cause spore deactivation while the addition of cyanide resulted in rapid spore deactivation. Treatment of activated spores with azide or environmental shifts also resulted in inhibition of oxygen uptake and spore deactivation. Deactivating spores did not demonstrate the amino acid incorporation, uridine incorporation, and expression of trehalase activity which is found in the later stages of germinating control spores. Protein synthesis inhibitors did not cause spore deactivation or a decrease in oxygen uptake but they inhibited amino acid incorporation and the expression trehalase activity in swollen spores. It is concluded that control of respiratory activity is involved in regulation of reversible activation.


Evaluation of an improved rapid coagglutination method for the serological grouping of beta-hemolytic streptococci. Grouping of beta-hemolytic streptococci was performed with the Phadebact Streptococcus Test, a coagglutination method, and the results compared with serological grouping by the standard Lancefield precipitin method. Of 171 clinical specimens examined, 169 (98.8%) were grouped correctly by the Phadebact Test after 24 h of continuous growth in Todd-Hewitt broth. In a parallel study, 96.9% of specimens that grew after only 4 h of incubation in broth were grouped correctly by the coagglutination method. In both studies, the accuracy of the coagglutination test was increased significantly by elimination of multiple-agglutination reactions through centrifugation of cultures and utilization of the supernatant fluid in the Phadebact Test.


[Identification of an extracellular nucleotide pyrophosphatase in the culture media of Streptomyces mediterranei ME/R 17]. An exocellular pyrophosphatase, active on the nucleotide precursors of peptidoglycans, has been found in the culture medium of Streptomyces mediterranei ME/R 17. This enzyme was separated from the DD-carboxypeptidase by batchwise adsorption on DEAE cellulose. The pyrophosphatase had no strict substrate requirements, it hydrolyzed various UDP-sugar substrates: UDP-GlcNAc, UDP-Mur NAc and UDP-MurNAc peptides, giving rise to the corresponding sugar phosphate and to UMP. The enzyme preparation also contained a 5'-nucleotidase activity and UMP was further split to give uridine. This nucleotidase activity was inhibited by potassium tetraborate. Both cytoplasmic and particulate preparations from cells of S. mediterranei also contained a pyrophosphatase activity while only the particulate fractions showed the DD-carboxypeptidase activity. The pyrophosphatase excretion was tested during the grwoth cycle. The activity of the enzyme showed a constant increase throughout the exponential growth and a stronger increase in the late exponential phase. Such a result could be correlated with a consumption of the nutrients in the culture medium, in fact a relatively poor culture medium had a strong positive effect upon the production of the exocellular pyrophosphatase.


Nitrogen nutrition and regulation of cephalosporin production in Streptomyces clavuligerus. When used as sole nitrogen source, certain amino acids (e.g., proline, asparagine) supported both growth and sporulation by Streptomyces clavuligerus streaked onto solid defined medium. Ammonium supported growth but suppressed sporulation. Amino nitrogen was best for cephalosporin production in liquid defined medium, although urea was almost as useful. A comparison of amino acids showed asparagine and glutamine to be the best nitrogen sources and arginine to be almost as good. Ammonium salts supported a somewhat lower growth rate than asparagine, but antibiotic production was very poor on these inorganic nitrogen sources. Addition of ammonium to asparagine did not affect growth rate but increased mycelial mass; cephalosporin production was reduced by about 75%. Antibiotic production was more closely associated with growth in the absence of ammonium than in its presence, indicating a strong inhibitory and (or) repressive effect of NH4+ on antibiotic production. Ammonium exerted its negative effect when added at 24h or earlier, i.e. before antibiotic formation began.


Fatty acid composition of Rhizobium spp. The fatty acid composition of 42 isolates belonging to the major plant affinity groups of Rhizobium has been determined and found to vary reproducible with culture age. Numerical taxonomic techniques applied to the 15 major fatty acid components of log-phase cultures of comparable physiological age showed that the rhizobia constitute a uniform group. However, two clusters comprising soybean-cowpea isolates and pea-bean isolates were evident. These observations, based on a simple analysis of only one group of chemical components, indicate relationships among rhizobia which differ from the conventional plant-affinity groupings but which are consistent with other proposed relationships established using a variety of biochemical and physiological criteria.


Ultrastructural studies on the cell walls in Fusarium sulphureum. The cell walls of Fusarium sulphureum have a microfibrillar component that is randomly arranged. X-ray-diffraction diagrams of the microfibrils are consistent with a high degree of crystallinity and show that they are chitin. The chitin microfibrils of the peripheral walls envelop the hyphal apex and extend across the septae. During the first 8h in culture, the conversion of conidial cells to chlamydospores is evidenced by a swelling of the cells and the original microfibrils remain randomly arranged. Within 24h new wall material is deposited as the cells expand and the wall thickens. The new microfibrils are indistinguishable from those of the original conidial cells. After 3 days in culture, the chlamydospores are fully developed and have the characteristic thick wall which is a continuous layer of randomly arranged microfibrils. Chlamydospores maintained in a conversion medium for 8 days have microfibrils identical with those in 3-day-old cultures; thus a further change in the microfibril orientation did not occur during that period. Alkaline hydrolysis of the walls removes most of the electron-dense staining constituents from the inner wall layer and leaves the outer wall layer intact. This treatment also reveals some of the wall microfibrils. An additional treatment of the walls with HAc/H2O2 completely removes the wall components that react positively to heavy metal stains. The results are discussed in relation to the structure of other fungal cell walls.


Temperature-sensitive mutants of bacteriophage SH-133 specific for the hydrogen bacterium Pseudomonas facilis: isolation, complementation, and partial characterization. Seventeen temperature-sensitive mutants of bacteriophage SH-133 have been isolated following mutagenesis with UV-light, nitrosoguanidine, and ethyl methanesulfonate. The mutants were classified into 15 complementation groups according to their ability to complement each other at 32 degrees C, the nonpermissive temperature. Each mutant was studied with regard to the relationship between its ability to multiply in heterotrophically (H-) and autotrophically (A-) grown Pseudomonas facilis cells. At 27 degrees C, the permissive temperature, the plaque-forming ability of the 17 mutants and wild-type phage was reduced 10-fold in A-grown cells. At 32 degrees C, mutants belonging to 10 groups exhibited identical levels of multiplicity-dependent leak under both modes of growth. However, the infection of A-grown cells by mutants belonging to the remaining five groups resulted in as much as 500-fold inhibition of multiplicity-dependent leak when contrasted with the infection of cells grown heterotrophically. These observations indicate that the expression of five SH-133 phage cistrons is defective when multiplication proceeds under autotrophic metabolism. Seven mutants were found to differ from the wild-type phage with regard to thermal stability at 56 degrees C which suggests that they possess altered structural proteins. Four of the seven thermosensitive mutants exhibited reduced levels of multiplicity-dependent leak in A-grown cells. The data suggest that the reduction in plaque-forming ability of SH-133 in A-grown cells is caused by a defect in the expression of specific phage structural components.


Evaluation of the body iron status of native Canadians. The serum ferritin concentration was measured in 1417 Indians and 310 Inuit aged 1 to 89 years. The subjects were initially selected to produce a representative sample of the entire native population, but the rate of nonresponse was high, and the results reported in this paper are representative only of the people studied.In males the median serum ferritin values increased during early life and tended to plateau after the age of 30 years. In females the median values rose during childhood, tended to plateau during adolescence, increased slightly during the reproductive period, then gradually rose thereafter. Ranges of values were wide in all age groups, reflecting the variations in body iron stores. When compared with the Inuit, the Indians had a significantly higher prevalence of abnormal serum ferritin values.From an analysis of the serum ferritin values in Indians it is probable that iron stores were reduced in approximately 30% of children, 40% of adolescents, 34% of nonpregnant women of reproductive age, 11% of older women and 5% of adult males. The corresponding figures for the Inuit were 15%, 23%, 22%, 6% and 1%. In contrast, iron deficiency anemia was found in only 3% to 4% of native peoples. If "normality" requires more than small amounts of iron stores to meet physiologic needs, the results suggest a high probability of iron deficiency in 20% to 40% of native children, adolescents and nonpregnant women of reproductive age, and in 0% to 10% of other subjects; but if "normality" is defined as adequate iron stores for erythropoiesis the prevalence of iron deficiency was approximately 1% to 2% in children and adolescents, 3% to 5% in women and less than 1% in adult males.


Factors associated with high-risk pregnancies in Canadian Inuit. Antenatal risk factors and management problems during labour and delivery were examined for 141 consecutive deliveries in Canadian Inuit of the eastern Arctic. The applicability of three risk scoring systems for antenatal use was evaluated. Pregnancies were categorized as being at high, moderate or low risk according to the authors' published criteria. Only two of the systems gave statistically valid predictions of the outcome of pregnancy in terms of risk categorization. Sensitivity and uniformity of the systems were lacking: the risk scores were low in 32% to 58% of the pregnancies in which the mother or the infant or both had problems during labour or delivery or both, and in 27% to 36% of the pregnancies in which the infant alone had problems. Antepartum factors that indicated a significantly increased risk of problems during labour or delivery or both were maternal age less than 16 or more than 35 years, prevous stillbirth or neonatal death, previous birth weight of less than 2501 g, previous pastpartum hemorrhage or other problem in the third stage of labour, antituberculosis therapy in the mother, and any of antepartum hemorrhage, multiple pregnancy, breech delivery, malpresentation or long period between rupture of the membranes and delivery in the current pregnancy. The data indicate that scoring systems should take into account regional or population variations if they are to have reasonable sensitivity.


A university department's involvement with medical care in the Canadian North. Both visiting consulting services and resident general practice services to the Churchill Health Centre in northern Manitoba are provided by the University of Manitoba through its northern medical unit. The roles of the university's department of obstetrics and gynecology with regard to visiting consulting services include patient care, education and medical audit. This paper reviews the several aspects of this involvement over the period 1971 through 1977.


Tuberculous meningitis in children: a retrospective study of 79 patients, with an analysis of prognostic factors. A study of 79 cases of tuberculous meningitis in children, with an emphasis on prognostic factors, was conducted by means of a review of hospital records and completion of a follow-up questionnaire. The survival rate was 62%. Stage 3 disease (with severe changes in the sensorium and often severe neurologic abnormalities) at the time of admission, age 3 years or less, associated miliary tuberculosis and delay in the initiation of therapy correlated significantly with poor outcome. Severe neurologic complications in the acute phase were also related to poor outcome and severe sequelae. General recommendations pertaining to treatment are made.


Parasitic disease in humans: the extent in Canada. Much is being said, often dramatically, about the potential hazards of parasitic diseases in Canada, but little or no attempt has been made to determine the true extent of the problem. Indigenous parasite pathogens are recognized in resident Canadians, and pathogens are acquired by travellers abroad or reported from immigrants. The role of each of these categories is important in the characterization of the problem of parasitic diseases in Canada. From data provided by provincial laboratories and hospitals it is estimated that 1 person in 1000 will spend 1 day per year in hospital because of intestinal parasites, and 1 in 100 each year will have a diagnosis of intestinal parasitic infection made from examination of a stool sample.


Autoimmune myocarditis: a clinical entity. In a case of myocarditis electron microscopic and immunoflourescent studies of a transmural myocardial biopsy specimen indicated an autoimmune process. Extensive inflammatory cell infiltration, immunoglobulin and complement deposition along the sarcolemma and in the interstitium, and capillary endothelial injury were found. After a short course of immunosuppressive therapy the inflammatory process was replaced by collagenous scarring and lymphocytic depletion; the blood vessels were then normal. Earlier therapy in such cases may be lifesaving.


Persistence of influenza serum antibodies in humans following immunization with a bivalent A/Victoria and A/New Jersey vaccine. The persistence of serum antibodies 1 year after immunization with a bivalent vaccine containing recombinant viruses that were antigenically identical with A/Victoria/3/75 (H3N2) and A/New Jersey/8/76 (Hsw1N1) viruses was measured in 128 persons aged 18 to 65 years. Serum samples were tested with the hemagglutination inhibition assay against the two vaccine antigens and against A/Texas/1/77 (H3N2) and A/USSR/90/77 (H1N1) viruses. Prior to vaccination 56% and 79% of the participants had been found to be seronegative to A/Victoria and A/New Jersey antigens respectively; the geometric mean antibody titres were low (1:5 to 1:11) except in persons aged 51 to 65 years, whose mean titre of antibody to the A/New Jersey antigen was 1:23, and persons aged 26 to 35 years, whose mean titre of antibody to the A/USSR antigen was 1:25. By 3 weeks after vaccination 85% of the seronegative persons had a fourfold or greater rise in titres of antibodies to the viruses in the vaccine, and 70% had a fourfold increase in titre of antibody to the A/Texas antigen. Of the persons aged 26 to 35 years (seronegative and seropositive) 68% had a fourfold or greater increase in titre of antibody to the A/USSR antigen. There was no change in the mean titres of 19 unvaccinated control subjects during the observation period. At 6 and 12 months after vaccination the titres of antibodies to the A/Victoria and A/New Jersey antigens had declined moderately in all age groups from those observed 3 weeks after vaccination. The rate of decline was similar for the various antibodies except that to the A/USSR antigen in persons 26 to 35 years of age, in whom the decline was much slower.


Porphyria cutanea tarda: clinical and laboratory features. Eleven patients with porphyria cutanea tarda were studied. Biochemical confirmation of the clinical diagnosis required only determination of the total urine porphyrin concentration in a sample of urine voided on rising in the morning. The patients were divided for convenience of discussion into four groups differing in age, sex and etiologic factors. Of the six patients in whom a liver biopsy was done one was shown to have micronodular cirrhosis. Except for a modest elevation in the serum glutamic oxaloacetic transaminase values when the patients were first seen, no evidence was found for liver disease apart from the presence of porphyria cutanea tarda. One patient recovered solely by abstaining from alcohol consumption. Five patients underwent phlebotomy; their iron stores had been found to be between 2 and 3 g. Decreasing urine porphyrin values correlated well with decreasing serum ferritin values during the course of phlebotomy. Porphyria cutanea tarda, which is due to a deficiency of uroporphyrinogen decarboxylase, is manifested in association with alcohol abuse, estrogen therapy, exposure to chlorinated hydrocarbons or increased tissue iron stores, or a combination of these factors. Although relatively uncommon, this condition raises important and unresolved issues regarding the hepatotoxicity of alcohol, estrogens, chlorinated hydrocarbons and iron.


Standardization of electrocardiographic interpretive statements: a menu for word processing. Standardization of electrocardiographic interpretive statements is a goal of various coding systems, but word processing has not usually been considered. A simple, easily memorized system for clinical electrocardiography has been developed and used for approximately 60 000 interpretations. It takes the form of a "menu", in which boxes stand for various interpretive statements; the boxes are identified by mnemonics and marked by the interpreter when appropriate. The results provide better standardization, significant decreases in the numbers of descriptive statements and words per interpretation and considerable saving in typing time. Acceptance by the interpreters has been good. Features of the system allow for word processing as part of a polarcardiography computing system.


Survey of the clinical use of pressurized aerosol inhalers. Questions as to the use of pressurized aerosol inhalers were asked of 130 persons presenting for routine pulmonary function studies who had used such a device in the previous year. After spirometry was conducted the subjects used the inhaler; they were unaware that their technique of use was being observed. Only 10.8% performed correctly all 11 maneuvers suggested for the proper use of the inhalers, and 24.7% failed to perform more than 5 satisfactorily. There were significant differences in performance according to referral diagnosis (chronic obstructive lung disease v. asthma) and regularity of use of inhalers.


From hospital to community: a system changes. Interest in the community and institutional aspects of mental health care in Saskatchewan goes back to the early history of the province, but it is the last 15--20 years which have seen the most exciting developments. During this time the province has changed from having one of the highest mental institutionalization rates in the world to one of the lowest. One mental hospital was phased out and the other reduced to a shadow of its former self. An account of the changes which occurred during this period is given together with the story of the stresses generated. Finally, a 1979 perspective is offered on the Saskatchewan psychiatric program.


A mental health system in transition: profiles of change. These profiles of the changing Saskatchewan mental health system reveal that there have been increases in the number of admissions, readmissions, discharges, and outpatients in both the public and private sectors. Divergent patterns in the utilization of services in the public sector occur among the Indian and non-Indian populations. Over the past several decades there have been considerable changes in the types of psychiatric problems treated in the psychiatric care system. The social, demographic and admission characteristics of the people receiving treatment in the system have changed substantially. The data presented here imply that the status of mental illness is not solely a function of a physical or psychiatric condition but has elastic features capable of being expanded or restricted by the prevailing organizational structure which has evolved to handle the "problem" of mental illness. The advent of community psychiatry plus attendant changes in the health system has altered the nature of mental illness treated in the province. Consequently, concerns about ways of evaluating the effectiveness of current programs, of choosing between alternate service delivery systems, of establishing criteria for equitable resource allocation, and of understanding the forces for change need to be raised and explicitly dealt with.


Relationship between high fasting blood sugar and depression in a mental hygiene clinic population. The relationship between diagnosis and abnormally high fasting blood sugar levels was reviewed for 4,994 patients in a mental hygiene clinic. Fifty-four patients exceeded the criterion level of fasting blood sugar abnormality, FBS greater than or equal to 145 mg%. There was no evidence of relationship between psychiatric diagnosis and the presence of an abnormally high fasting blood sugar level. This held true for the diagnosis of depression despite observations made elsewhere that a significant association exists between the depressions and diabetes.


Suicide in Newfoundland and Labrador.

Suicide in Newfoundland and Labrador. The present investigation was undertaken to explore the special factors associated with low suicide rates in Newfoundland and covered the ten year period of 1964 to 1973. The data were collected from death certificates. Comparisons were made with figures for Canada as a whole, as reported in the official statistics. Apart from a constant overall low rate of suicide in Newfoundland, women and young people (aged 15 to 29) failed to show an increase in the suicide rates such as has been observed in other parts of Canada and throughout the western world. In Newfoundland, age specific suicide rates, choice of method, variations in the rates for different religious denominations and seasonal variations in suicide rates were different from those observed elsewhere. Generally, suicide in Newfoundland shows a much older pattern and does not involve the new high-risk groups seen in more urbanized societies. Possible explanations for the special demographic characteristics of suicide in Newfoundland are offered, mainly on social, cultural and geographical grounds.


The mystic experience: a psychiatric reflection. This paper reviews the phenomenology of the mystical experience and its varied contexts epilepsy, toxicity, organic brain syndromes, the major psychoses and hysterical dissociative states as well as in apparently normal persons. The impact of the experience on the personality is noted and its significance briefly reviewed. The author notes that two fallacies await the unwary psychiatrist: the fallacy of reductionism which defines the mystical experience in pathological terms only; and the fallacy of speculation without adequate philosophical or theological tools.


Anorexia nervosa: death from complications of ruptured gastric ulcer. Death from the complications of a ruptured gastric ulcer in a 15 year old girl with anorexia nervosa is reported. A Medlars II search of the literature indicates no previously reported deaths from this cause in anorexia nervosa. This patient presented the classical clinical characteristics of the syndrome and the course of her illness serves to re-emphasize the seriousness of the disease and the importance of early management of anorexia nervosa.


Hysteria: a case note study.

Hysteria: a case note study. In a matched retrospective study, 31 patients with hysterical neurosis were compared with 31 with depressive neurosis. There were no significant differences between the groups for siblings' position, medical work, brain disease, poor marriage, frigidity, or family history of psychiatric disorder. Significantly more of the hysterics had a preceding head injury and 29% had a past history of hysteria. By the time of the study 48% had been treated for depression.


The research diagnostic criteria: historical background, development, validity, and reliability. This article reviews the theoretical papers and research studies concerning the Research Diagnostic Criteria (RDC), a research tool used to group psychiatric sugjects into homogenous diagnostic groups. The RDC contains specified inclusion and exclusion criteria for 25 diagnostic categories. The contributions made by the Feighner and Spitzer groups in the field of diagnostic reliability and validity are briefly outlined. The development of the RDC and studies regarding its reliability and validity are presented and discussed. A review of the reliability studies, which basically employed an interrater design, indicated that the RDC is a relatively reliable diagnostic tool, obtaining interrater kappa coefficients which are consistently higher than those obtained, for similar categories, in studies using other diagnostic systems such as the DSM-II. Very few studies, however, directly investigated the validity of the RDC. Its assumed validity is dependent on the findings from studies which used similar diagnostic criteria. It was suggested that more studies would be required to establish the validity of the RDC. Several limitations of the RDC were pointed out. Included among these were: its use of an exclusion method to determine diagnostic criteria, the difficulty and great amount of time required for its use, and the fact that it has so far been used only with adult inpatients. Despite its limitations, it was concluded that the RDC remains one of the best tools available to a researcher who wishes to group psychiatric patients into homogenous groups, and that the clinician could advantageously use it in his clinical practice.


Myopathy and myoglobinuria in feedlot cattle. Myopathy and myoglobinuria occurred in a group of feedlot cattle in Alberta following transportation. The condition was associated with a deficiency of selenium (Se) in grain and hay fed to these animals on their premises of origin. Clinical findings in affected animals were characterized by a normal body temperature, stiffness, sternal recumbency and the passage of red or coffee-colored urine. Treatment with injectable Se-vitamin E was followed by the recovery of most affected animals within two weeks.


Feline urological syndrome.

Feline urological syndrome. A case of feline urolithiasis is described. It was demonstrated that serial determinations of renal function were mandatory in order to provide a favorable prognosis. Because hyperkalemia was present, the fluid of choice for the initial treatment was saline. The various considerations in the prevention, treatment and monitoring of cats with feline urolithiasis syndrome are discussed.


Bovine granular vulvitis associated with ureaplasma infection. A granular vulvitis syndrome associated with ureaplasma infection was first recognized in Ontario dairy herds in 1972. The acute form of the disease was characterized by a purulent vulvar discharge, an inflamed hyperemic vulvar mucosa and varying degrees of granularity. In the chronic form, there was an absence of a purulent discharge and a gradual decline in the severity of the hyperemia and granularity. Epithelial inclusion cysts were observed in the vulvar epithelium of approximately 10% of affected cows.A seasonal variation in the incidence of the disease was observed. Herd morbidities during the summer months reached a low of 37% and increased to 75% during the winter months with constant housing.When widespread in herds, the acute form of the disease had a significant effect on fertility. In four herds examined, first service conceptions dropped on average by 27%. The chronic form of the disease had a less detrimental effect on fertility with first service conceptions being reduced on average by 13%. Intrauterine infusions of a tetracycline 24 hours postbreeding were found to be of value in improving conception rates in acutely affected herds.


A slim 198gold-grain implanter loaded with standard Royal Marsden 14-grain magazines. We designed a slim gold-grain implanter with adaptable lengths to implant areas accessible only through long, narrow, examining instruments, such as a suspension laryngoscope. The implanter is loaded with the same 14-grain magazine designed for and supplied with the Royal Marsden "gun." The simplicity of the loading mechanism with a minimum of moving parts makes the instrument practically trouble free. Although it is designed to be used along narrow examining instruments, it can also be used in any situation in which a permanent implant is required, for instance, prostatic cancer and pelvic recurrences in cancer of the uterine cervix previously treated by external and intracavitary irradiation.


Treatment of endometrial cancer in a regional radiation therapy center: analysis of 379 consecutive patients. Three hundred seventy-nine consecutive patients with adenocarcinoma of the corpus uteri, in whom irradiation was part of the treatment, were reviewed. The minimum follow-up was 3 years and none were lost. An increased incidence of corpus cancer in Central New York was demonstrated but could not be attributed to estrogen usage. There were 310 patients in FIGO stage I and 40 in stage II. Adenocarcinoma was reported in 319 and adenoacanthoma in 45. Overall survival was 74% and 62% at 5 and 10 years, respectively, and in stage I, 82% and 68%, respectively. Results for adenoacanthoma were similar to those for adenocarcinoma. Ten year survival was significantly better in those patients with a histologically negative hysterectomy specimen after preoperative intracavitary irradiation than in those with residual cancer. Treatment was tolerated well. This study demonstrates the results which can be obtained in a regional radiation therapy center serving patients and gynecologists from a wide geographic region operated upon in many different hospitals.


Conservation surgery and irradiation for the treatment of favorable breast cancer. The results in 162 patients with clinically favorable breast cancer treated with conservation surgery and radiation therapy are presented. The surgical procedures were simple excision with and without positive microscopic margins, segmental mastectomy, and segmental mastectomy with axillary dissection. Details of the radiation techniques are described with an explanation of the modifications in technique depending on the prior surgical procedure. Excellent control of local and regional tumor (96%) gives support to the combined treatment without removing the breast.


The place of interstitial therapy using 192 iridium in the management of carcinoma of the lip. We present the results of interstitial therapy with 192 iridium for 91 patients with carcinoma of the lip. This technique, without complications or sequelae, assures the control of local disease in 95.5% of cases (87/91). The management of metastatic lymph nodes and the indications for neck dissection are discussed.


Hodgkin's disease, stages I and II: relationship of recurrence to size of disease, radiation dose, and number of sites involved. A total of 57 patients with Hodgkin's disease limited to above the diaphragm (Stages I and II, A and B) were treated with radiation therapy alone at the University of Florida between 1964 and 1974. Staging laparotomy was done on 24 patients (42%). A total of 16 patients have relapsed, 4 within the treatment fields, 3 marginally, and 9 as extensions. Analysis of dose vs. size showed no in-field failure with disease less than 6 cm in size, with a range of doses. Massive disease continued to recur occasionally, even with higher doses. Most of the massive disease was mediastinal or hilar; the risk of lung or pleural involvement as determined by x-ray and/or biopsy showed a dramatic increase with size of mediastinal/hilar disease. Only 2 (5%) of 43 patients with mediastinal disease less than 6 cm in size had lung or pleural involvement, compared with 10 (71%) of 14 patients with disease greater than 6 cm in size. Both in-field recurrence and generalized relapse were closely related to the number of anatomical sites involved. Treatment programs need to consider tumor size, particularly in the mediastinum, and the number of sites involved.


Combined chemoimmunotherapy for advanced breast cancer: a comparison of BCG and levamisole. One hundred and fourteen evaluable patients with metastatic breast cancer were treated with a program consisting of 5-FU, Adriamycin, cyclophosphamide (FAC) and nonspecific immunotherapy with Levamisole. The results of this treatment program were compared to those observed with FAC and Bacillus Calmette Guerin (BCG) and FAC chemotherapy alone, both groups treated prior to the study reported in this paper. The overall response rates and complete response rates for all three treatment regimens were identical. The duration of remission, survival of all patients and survival of responders was similar for both chemoimmunotherapy regimens, being superior to the FAC chemotherapy alone group. Immunotherapy with Levamisole was well tolerated and side-effects were experienced by less than one-fourth of the patients. Overall, Levamisole was better tolerated than BCG and was easier to administer than the latter drug. These results suggest than nonspecific immunotherapy with Levamisole might prolong remission and survival of patients with metastatic breast cancer. Since the results achieved with BCG and Levamisole appear similar, the therapeutic ratio favors the use of Levamisole.


Small cell tumor of cervix with neuroepithelial features: ultrastructural observations in two cases. Two cases of small cell tumor of the uterine cervix are reported in which electron microscopic studies indicated that the tumor cells were of neural crest origin. Both tumors behaved aggressively, and metastatic involvement of pancreas was demonstrated in one patient, while the second patient developed liver metastasis. The need to distinguish these neoplasms from squamous cell carcinoma of the cervix is stressed. The tumors are compared with small cell neuroepithelial neoplasms previously reported as adult neuroblastomas.


Hepatic Hodgkin's disease without involvement of the spleen. A patient with Hodgkin's disease, who at laparotomy had liver involvement with no evidence of Hodgkin's disease in the spleen, is reported. Rare alternate lymphaticovenous anastomoses exist that may lead to liver involvement in the absence of splenic involvement.


Desmoplastic malignant melanoma: A light and electron microscopic study of two cases. This light and electron microscopic study of two recent cases of desmoplastic malignant melanoma (DMM) attempts to resolve the conflict in views regarding the nature of the cells responsible for the desmoplasia associated with this clinicopathologic entity. On the basis of evidence presented, it is concluded that the cells are dedifferentiated tumor cells with fibroblastic features and probably functions, rather than host engendered fibroblasts in response to invasive melanoma. The evidence includes: observation of macular desmosomes between tumor cells, an unheralded feature previously noted in amelanotic and melanotic melanomas; electron microscopic observation of fibroblast-like cells by others in spindle cell squamous carcinomas; and light microscopic features of malignancy including vascular invasion in one of the two cases. A reproducible light microscopic pattern diagnosis of this variant of malignant melanoma is reaffirmed in both cases.


Dysgerminoma of the ovary.

Dysgerminoma of the ovary. From 1938 to 1977, 21 patients with pure dysgerminoma were treated at Memorial and James Ewing Hospitals, now Memorial Sloan-Kettering Cancer Center. Twelve patients were considered to have their initial therapy begun or completed at our institution. Eleven (91.7%) of 12 patients were found free of disease from 2 to 38 years (median 6 years). Nine patients were referred with recurrent disease. Only 4 (45%) could be salvaged 2 to 21 years (median 9 years). Because of the lower survival rate in the treatment of recurrent disease in this series, it is difficult to advocate overly conservative therapy initially in all early lesions. The survival rate depends on the original size of the tumor, initial adequate staging and appropriate therapy based on the extent of disease. Recommendations for a treatment protocol are proposed.


Risk factor analysis in differentiated thyroid cancer. Six hundred patients with primary differentiated thyroid carcinoma had follow-up studies for a minimum of 15 years and a maximum of 45 years. Recurrence rate and death rate were significantly different in defined high-risk and low-risk groups of patients. These basic risk groups were defined by age and sex alone; low risk consisted of men 40 years of age and younger and women 50 years of age and younger whereas the high-risk group were older patients. Recurrence and death rates in patients at high risk were 33% and 27% while respective figures for patients at low risk were 11% and 4%. In more recent years these results have shown significant improvement. Basic risk group definition outweighed the effect of pathologic type, local disease extension, type of treatment, and site of recurrence or metastasis. For instance, radioactive iodine cured 70% of patients at low risk with metastatic disease but only 10% of patients at high risk. Less aggressive biologic  behavior of thyroid cancer before the age of menopause implies that an estrogen-rich milieu may alter the effects of initiating and promoting factors in carcinogenesis. It also suggests that therapeutic trials of estrogen be undertaken in progressive metastatic differentiated thyroid cancer.


Treatment of intraoral cancer by electrocoagulation. Electrocoagulation of intraoral carcinoma has been utilized as primary curative treatment in a series of twenty-five patients. The technique described here employs a disk electrode, requires only light general anesthesia and has a low rate of complications. Eleven patients survived more than five years. Eight died of other diseases and only 6 died with recurrent cancer. This is an effective technique which deserves wider use.


Medullary thyroid carcinoma: importance of serial serum calcitonin measurement. Thirty-two patients with medullary carcinoma of the thyroid (MCT) were studied before and after therapy. Sixteen patients had familial and 16 had the sporadic type of disease. The groups differed in several features: Patients with sporadic disease were older at diagnosis and presented with enlargements in the thyroid or lymph nodes, and one patient had neuromata; in the familial group the disease was more often bilateral and was associated at times with the MEA syndrome (parathyroid adenoma or pheochromocytomas, or both). The immunoreactive serum calcitonin (CT) level was measured before and after some form of therapy in all 32 patients. Our investigations showed: 1) Delaying treatment was clearly detrimental in this progressive disease; 2) The most effective therapy was surgery, while radiotherapy and chemotherapy were less effective; 3) Postoperatively, the CT level usually fell sharply, when the disease was thought extirpated, but the lowest nadir might be reached at from 1 month to 6 years; 4) In all such patients the CT level eventually rose from the postoperative nadir; 5) Patients with no clinical or radiological evidence of disease, had high CT levels for a mean of 3 years of observation; 6) A marked rise of CT levels in three patients preceded metastases and death; 7) Patients with abdominal, particularly liver disease had the highest CT levels; and 8) Patients who died had relatively high CT levels, corresponding in general with the extent of disease.


Diagnosis of minimal breast cancers in the BCDDP: the 66 questionable cases. Of 1,810 breast cancers detected in the NCI/ACS Breast Cancer Detection Demonstration Projects, 592 were less than 1 cm in diameter and considered minimal; and tissue slides from 506 of them were available for retrospective review by a panel of pathologists. The initial report of this review indicated that in 66 cases the pathologic features of the presented slides were not sufficient for diagnosis of cancer. Subsequent investigation revealed that, through computer error, the slides submitted in 2 of these 66 cases were not from the lesions in question but from blind biopsy of the contralateral breast. Further review by the pathology panel of tissue from 38 of the 64 remaining cases determined that 16 of the remaining lesions were indeed cancers or borderline malignant lesions. This then left 48 cases in doubt. Only biopsy had been performed in 11 of them, and some form of mastectomy in the other 37. The original pathologic opinion had been divided in 30 of these, and the mastectomy had been delayed for 1 day to 7 months after the biopsy. In only 7 of the 48 questionable cases was definitive treatment carried out at the time of biopsy. All in all, these findings reflect sound, responsible surgical judgment.


Relation between mammary cancer growth kinetics and the intervals between screenings. The purpose of this study was to consider the time interval for periodic mammographic screening for breast cancer. One hundred fifteen breast cancers occurring in 10,128 women receiving over 30,000 mammograms over a four year period were reviewed. Tumors were diagnosed at three time intervals: 1) first screening (39/115); 2) annual examination (27/115); and 3) at an examination that occurred less than twelve months from a previous annual examination (10/115). Also, there were tumors that grew to a palpable dimensions and were self-detected between annual examinations (39/115). Our opinion is that screening intervals should be individualized to each patient according to risk factors and suspicious mammographic findings. Further, there is a significant number of breast cancers that grow too fast to be detected effectively by annual mammography. Suspicious mammographic findings did not exist before these cancers reached palpable dimensions. Other risk factors characterizing the hosts who develop these fast growing cancers are yet to be determined.


Mammography in symptomatic women 50 years of age and under, and those over 50. Our experience with the use of mammography as a diagnostic aid in symptomatic women with breast complaints has been presented. One-third of all cancers were found in women 50 years of age and under. The detection of cancer by mammography in the younger age group was similar to that experienced in the older age group. Radiation exposure by mammography has diminished markedly during the last 15 years--only one-third to one rad per exposure with modern technique. Recent hysterical criticism of the use of mammography in women under 50 years of age is unwarranted in the light of current findings. This diagnostic modality should be utilized when indicated in all age groups over 30. It is probable that screening clinics should include all women over 35 years of age.


Serum-mediated immunosuppression in lung cancer. Immunosuppression in 45 patients with lung cancer was studied by examining delayed cutaneous hypersensitivity reactions to DNCB and by analyzing the effect of the patient's serum on the proliferative response of normal donor lymphocytes. Both diminution of DNCB reactivity and inhibition of the proliferative response of normal donor lymphocytes to mitogens were associated with the stage of the disease and the presence of unresected tumor. Suppressive sera were associated with poor prognosis. The suppressive effects of patients' sera on lymphocytes from a normal donor suggest that the immunosuppression seen in lung cancer may be mediated by serum factors. The significant association of clinically evident tumor with this serum-mediated immunosuppression further suggests that the tumor itself could account for the appearance of these factors in the host. The clinical implications of these findings may be useful for designing new clinical trials.


The quantitative and qualitative impairment of wound healing by adriamycin. Clinical impression suggests that Adriamycin (ADR) interferes with wound healing. To examine the effects of ADR on wound healing, male Fischer rats were subjected to a dorsal, midline, full-thickness longitudinal incision (day 0). Wound clips were removed on day +7. Twenty animals per group were given intravenous ADR on day -7, day 0, day +3 and day +7. Twenty animals served as non-treated, wounded controls (C). Five animals/group were sacrificed on days +7, +14 and +21, at which time two 9.5 mm wide strips were taken from each animal perpendicular to the wound axis and submitted for wound breaking strength (WBS) measurements and load-extension curve analysis. WBS differed most markedly at Day 21 between C(1671 +/- 59g) and ADR day -7(1360 + 71 g) p less than 0.01; C and ADR day 0 (1051 +/- 108 g) p less than 0.001; C and ADR day + 3(1134 +/- 176 g) p less than 0.02. No difference existed between C and day +7 (1790 +/- 153 g). A point of inflection always occurred between 55-60% elongation in ADR treated animals only. This portion of the curve has been previously shown to represent collagen content. It is concluded that perioperative ADR administration (day -7 through day +3) significantly and substantially impairs skin wound healing in the rat. A form of collagen yielding underlies and may contribute to this defect.


Multiple primary melanoma.

Multiple primary melanoma. From a series of 712 patients with melanoma, 38 patients (5.3%) had more than one primary melanoma. Twenty-four patients had two primaries, 11 patients had three, 2 patients had four, and 1 patient had eight. Twelve patients (32%) had one or more synchronous primaries. Forty-five percent of all multiple primaries were diagnosed within the first year. Microstaging by level and depth was determined prior to treatment and in patients with nonsynchronous primaries, 83% had a subsequent melanoma equal or less advanced than the original. Twenty-six patients with Stage I primaries were skin-tested with DNCB prior to therapy. No significant differences in delayed cutaneous hypersensitivity reactions were found between multiple primary and matched controls with only a single melanoma. Four of 10 patients with multiple primaries treated with adjuvant BCG or BCG-tumor cell vaccine developed subsequent melanomas suggesting that immunotherapy with BCG will not prevent the development of a new primary melanoma. Survival in patients with Stage I and II multiple primary melanomas was improved compared to Stage I and Stage II patients with a single primary. This study suggests that prognosis in multiple primary melanomas is better reflected by the most advanced primary based on microstaging and the presence or absence of regional lymph node metastases than by multiplicity.


Immune impairment and metastatic tumor growth: the need for an immunorestorative drug as an adjunct to surgery. A spontaneous murine metastatic tumor system was used as a model to assess the effects of a major surgical procedure on tumor-specific immune reactivity and the growth of micrometastases. Any major surgical procedure resulted in impaired cell-mediated cytotoxicity postoperatively and an increase in the number of gross pulmonary metastases. The use of an immunorestorative drug, Thiabendazole, in the perioperative period resulted in an improved cytotoxic response and a significant decrease in pulmonary metastases. Perioperative immunotherapy can be an effective adjunct to surgery in preventing the growth of micrometastatic foci.


The results of treatment of perineal recurrence of cancer of the rectum. A study of 36 patients with adenocarcinoma of the rectum recurrent in the perineum after Miles abdominoperineal resection defined a subgroup of 21 patients in whom curative re-excision of the perineum, with or without associated resection of viscera as warranted by operative findings, was performed. Recurrence without palpable disease was disclosed microscopically in seven patients (33%). None of the 21 patients had clinical or laboratory evidence of dissemination at the time of re-excision. Mortality and morbidity were low and yield in terms of pain control was excellent. Results achieved in patients treated with pre- and/or postoperative radiotherapy did not differ quantitatively or qualitatively from those treated by operation alone. Survival free of disease proved to be disappointing and underlines at present the palliative nature of this approach when the primary operation has been an adequate standard abdominoperineal operation. This report provides further evidence as to the value of the better definition of high risk primary rectal cancers and the need for continuing refinement of the follow-up and retreatment process.


Symptomatology as an indicator of recurrent or metastatic breast cancer. Eighty-seven patients with recurrent breast cancer after mastectomy were analyzed for patterns of recurrence and methods of detection. After an average disease-free interval of 30 months, 38% developed osseous metastases, 16% recurred locally, 10% had local plus systemic disease, 10% showed pulmonary metastases and the remainder were distributed among liver, brain, and remaining breast disease. In 79 patients recurrence was heralded by symptoms. Physical examination in five asymptomatic patients revealed local or supraclavicular recurrence. In only three asymptomatic patients was recurrence documented by "routine" chest x-rays (in two), or liver enzymes/liver scan (in one). No asymptomatic disease was found by bone scan. It is concluded that periodic history, physical examination, and chest x-rays are the most important components in the follow-up of breast cancer patients. Radioisotope scans and other radiographs are valuable in confirming symptomatic disease and detecting additional diseases, but cannot be recommended routinely in the asymptomatic patient because of low yield and cost.


Staging of cancer of the colon and cancer of the rectum. A retrospective analysis of 1,826 cases (924 colon, 902 rectal) from ten institutions provided the basis of this study on the staging of cancer of the colon and rectum. The general rules of the American Joint Committee on the relationship between times and the staging of cancer have been followed. These represent modifications of the originally formulated TNM system of the Union Internationale Contre Le Cancer (UICC) which has been designed as a clinical-diagnostic classification, not applicable to cancer of inaccessible sites or structures requiring postsurgical treatment pathologic assessment of therapeutically removed specimens. Inadequacies of the clinical data requested for our study required adoption of the pTNM evaluation method of classification. Multiple regression analysis of the data demonstrated a relationship between survival and the following: depth of penetration (T), status of regional lymph nodes (N), and presence or absence of distant metastasis (M). This was similar for both sites. Basically, for the rectum it was in consonance with the original Dukes' classification (A, B, and C), and was remarkably applicable to the colon. The survival data for the two sites were so similar as to suggest the use of one set of pTNM categories not only for the postsurgical-treatment pathologic evaluation, but also for the stage grouping definitions. Strongly recommended for cancer of all sites is the development of General Oncology Data Forms to be included in the clinical charts and records of all patients with cancer.


Steroid hormone receptors in human colon cancers. Tumors from patients with primary colon cancer were studied for the presence of steroid hormone receptors for estrogen (E2), progesterone (Prog), dihydrotestosterone (DHT) and glucocorticoid. Ten of 33 (30%) tumors contained high affinity E2 receptors. Four were males and six females with positive assays predominantly from the left colon. Twenty-three of these tumors were also assayed for DHT and Prog and six (26%) contained all three receptors. An additional twelve tumors had at least one receptor, so that 70% of the tumors studied contained one or more receptors. Five of 22 (23%) samples were positive for glucocorticoid receptors. Common etiological factors associated with colon and breast cancer were briefly discussed. These factors, along with the presence of hormone receptors in primary colon malignancies suggest that some large bowel cancers may be endocrine-dependent.


Development of a metastatic brain tumor model in mice. This study reports an easily accomplished and reliable model of metastatic tumor in the brains of mice. Five experimental groups of female C3H/Bi mice received left intracardiac injections of a syngeneic KHT sarcoma cell suspension (1 X 10(5) cells) and were followed until death. Two groups of mice also received 3,000 rads of radiation to a limited cardiac port 24 hr after tumor injections. All mice were completely autopsied, and the brains were examined both grossly and microscopically. Metastatic brain tumor developed in 60 to 70% of mice; the tumor foci were parenchymal, usually multifocal, and had wide distribution throughout the cerebrum, brainstem, and cerebellum. There was occasional meningeal tumor, but tumor never involved the skull, choroid plexus, pituitary gland, or local extracranial structures. Cardiac irradiation did not increase the number of the mean survival of mice with metastatic brain tumor but did decrease the total tumor burden of individual animals by markedly reducing the incidence of metastatic lung tumor and totally preventing tumor infiltration of the heart. This demonstration of consistently produced blood-borne metastatic brain tumor in mice should provide a valuable research model which will allow the central nervous system to be studied for internal mechanisms and/or external factors which influence the arrest and growth of embolic tumor cells in the brain.


N,N-dimethylformamide-induced alteration of cell culture characteristics and loss of tumorigenicity in cultured human colon carcinoma cells. Human colon carcinoma cell lines established in this laboratory were treated in vitro with N,N-dimethylformamide. This polar solvent caused morphological changes in the cells as well as alterations in their growth properties. Untreated cells had cloning efficiencies of up to 77% in soft agar; treatment with N,N-dimethylformamide resulted in a complete loss of clonogenicity in semisolid medium. Growth in the presence of the polar solvent also effected a marked reduction in the tumorigenicity of the cells. Ten of ten nude mice that received a s.c. inoculum of 1 X 10(6) untreated cells developed tumors histologically similar to colonic adenocarcinomas in 10 to 14 days, whereas nine of ten nude mice inoculated with 1 X 10(6) treated cells have shown no sign of tumor 3 to 6 months postinjection. Removal of the polar solvent from the culture medium was accompanied by the reappearance of tumorigenicity and the original cell culture characteristics. Therefore, it appears that N,N-dimethylformamide can reversibly effect the reversion of cultured human colon carcinoma cells to less malignant cell types.


A survey of growth in soft agar and cell surface properties as markers for transformation in adult rat liver epithelial-like cell cultures. Continuous epithelial-like cell lines derived from normal adult rat liver and hepatocarcinomas were evaluated for their growth in soft agar and five properties of the cell membrane as markers for neoplastic transformation. A correlation of these properties was made to the tumorigenicity of the lines in nude mice. Growth in soft agar was a specific and sensitive marker, whereas the data on uptake of 2-deoxy-D-glucose were consistent, with high uptake being a specific but clearly not a sensitive marker. Agglutination and hemadsorption mediated by concanavalin A, multinucleation in the presence of cytochalasin B, and the cell membrane activity of adenosine triphosphatase did not correlate with tumorigenicity of the other markers for transformation. In addition, it is shown that Mycoplasma infection does not alter any of these properties but that infection can be eliminated by passage of cells through nude mice.


Comparative studies of glucose metabolism in HTC, RLC, MH1C1, and Reuber H35 rat hepatoma cells. Carbohydrate metabolism by four rat hepatoma cell lines in culture, namely, Reuber H35, MH1C1, RLC, and HTC, has been investigated. Glucose utilization by H35 and MH1C1 cells is lower than that by RLC and HTC cells. The four cell lines also differ with respect to the accumulation of lactic acid in the growth medium; in particular, H35 cells show uptake of lactic acid, rather than accumulation in the medium. Specific activities of a number of enzymes involved in glycolysis, gluconeogenesis, pentose phosphate pathway, and glycogen formation were determined in the four cell lines. A direct relationship between the differences was found for the activities of some enzymes belonging to carbohydrate metabolism, namely, hexokinase, pyruvate kinase, aldolases A and B, glucose-6-phosphate dehydrogenase, and phosphogluconate dehydrogenase and the differences found for glucose utilization by the different cell lines.


Induction of differentiation of Rauscher virus-induced mouse myeloid leukemia cells with a factor(s) in ascitic fluid and inhibitors of nucleic acid and protein syntheses. Cell line R453, established from a Rauscher virus-induced myeloid leukemia in a C57BL/6 mouse, was induced to differentiate in vitro into macrophages and granulocytes with ascitic fluids from animals bearing various ascites tumors or from mice treated with complete Freund's adjuvant, conditioned media from various cell lines, and glucocorticoid hormone. Differentiated R453 cells had a morphology similar to that of macrophages and granulocytes in normal hematopoietic organs, and they phagocytized small paricles such as latex particles, moved in soft agar showing locomotive activity, and had Fc and C3 receptors on the cell surface. This induction of differentiation of R453 cells was markedly enhanced by addition of inhibitors of RNA synthesis (actinomycin D, nogalamycin, or chromomycin A3), protein synthesis (puromycin or cycloheximide), or DNA synthesis (methotrexate, hydroxyurea, 5-fluorodeoxyuridine, or 1-beta-D-arabinofuranosylcytosine) in the presence of ascitic fluid. Of the inhibitors, actinomycin D was the most effective at a low concentration (5 ng/ml) in stimulating induction of differentiation of R453 cells. However, these inhibitors alone did not induce differentiation of R453 cells. The factor(s) in ascitic fluid that stimulates differentiation of R453 cells was heat labile, nondialyzable, and inactivated by trypsin.


Synergistic action of vincristine and adriamycin in the treatment of experimental rat leukemia L5222. The therapeutic effect of different ratios of a combination of vincristine and Adriamycin was compared to monotherapy with both substances in advanced rat leukemia L5222. The combination therapy proved to by synergistic. In the upper dose range, no additive toxicity was observed. The combination of 25% vincristine plus 75% Adriamycin effected the best results. In a clinically relevant dose range, the superiority of this ratio was observed independent of the application sequence of the two substances and independent of the time interval between the applications within an 11-hr investigation period.


Metabolism and formation of DNA adducts of benzo(a)pyrene in human diploid fibroblasts. Cultured human diploid skin fibroblasts incubated with [G-3H]benzo(a)pyrene yielded about 10 times more H2O-=soluble benzo(a)pyrene metabolites and DNA adducts of stationary growth phase than did proliferating cultures. This increased formation could be blocked by alpha-naphthoflavone. Trichloropropenoxide and cyclohexenoxide, inhibitors of the epoxide hydratase, inhibited predominantly the formation of DNA adducts. Cultures from older individuals formed significantly more benzo(a)pyrene metabolites and DNA adducts, but control cultures from patients with either lung cancer or melanoma did not. The age influence was not apparent when the ratio of DNA adducts to H2O-soluble metabolites was determined for each individual cell line. However, the proportion of DNA-bound material in the cells from patients with lung cancer was significantly increased compared to cells from melanoma patients or healthy individuals.


Early decrease in hyaluronidase-sensitive cell surface charge during the differentiation of Friend erythroleukemic cells by dimethyl sulfoxide. Early membrane events in erythroid differentiation were investigated by means of cell electrophoresis utilizing cultured Friend erythroleukemia cell clones of different inducibility. The cell electrophoretic mobility decreased by 18% within 30 min of treatment with 1.5% dimethyl sulfoxide (DMSO) in highly inducible clones but not in noninducible clones. The reduced mobility persisted for 5 days of incubation with DMSO until hemoglobin synthesis. DMSO treatment for less than 16 hr and subsequent incubation without the drug resulted in the complete recovery of the mobility and no hemoglobin synthesis. Longer exposure to DMSO resulted in the loss of recovery of mobility and an increasing fraction of benzidine-positive cells seen on Day 5. Measurement of the electrophoretic mobility after the removal of acidic sugars by their specific enzymes suggested that hyaluronidase-sensitive negative charges were lost from the cell surface only in highly inducible clones. The mobility reduction associated with hyaluronic acid was also caused by other potent inducers (sodium butyrate, N-methylacetamide, and N,N-dimethylacetamide). These results suggest that the decrease in cell surface glycocalyx might be an early step in the induction of differentiation of Friend erythroleukemia cells.


Stability of estrogen receptors in frozen human breast tumor tissue. The effect of freezing on the stability of the estrogen receptors in human breast tumors was measured. Portions of three tumors kept frozen for 3 days after removal from the patients were analyzed by the sucrose density gradient method. The initial measurements demonstrated positive receptor values. The remaining portions were kept frozen for 30 days, and the receptor values were reevaluated. The 8S receptor was 80% destroyed under these freezing conditions. The 4S receptor was found to be more stable. The inactivation reflects a loss of specific binding since the protein concentration was similar in the analyses. These results strongly indicate that human breast tumor specimens should be analyzed as soon as possible or within 3 weeks after freezing.


Metabolism of aflatoxin B1 and identification of the major aflatoxin B1-DNA adducts formed in cultured human bronchus and colon. Aflatoxin B1 and benzo(a)pyrene were activated by both cultured human bronchus and human colon as measured by binding to cellular DNA and protein. The binding of aflatoxin B1 to DNA was dose dependent, and the level of binding was higher in cultured human bronchus than it was in the colon. When compared to aflatoxin B1, the binding level of benzo(a)pyrene to both bronchial and colonic DNA was generally higher. The major adducts formed in both tissues by the interaction of aflatoxin B1 and DNA were chromatographically identical to 2,3-dihydro-2-(N7-guanyl)-3-hydroxyaflatoxin B1 (Structure 1) with the guanyl group and hydroxy group in trans-position and an adduct which has been tentatively identified by other investigators as 2,3-dihydro-2-(N5-formyl-2',5',6'-triamino-4'-oxo-N5-pyrimidyl)-3-hydroxyaflatoxin B1 (Structure 11). Seventy % of the radioactivity associated with bronchial DNA was found in these two peaks, and the ratio of radioactivity between the peaks was nearly 1. In colonic DNA, the ratio between Structures 1 and 11 was approximately 2. These observations add aflatoxin B1 to the list of chemical procarcinogens metabolized by cultured human tissues and in which the carcinogen-DNA adducts are similar to the adducts formed in animal tissue susceptible to the carcinogenic action of aflatoxin B1.


Metabolism and effects of 5-(beta-D-ribofuranosyl)isocytosine in P815 cells. 5-(beta-D-Ribofuranosyl)isocytosine (psi l Cyd), a C-nucleoside, has been shown to be active against P815 leukemia in mice. In P815 cells treated with [2-14C]psi l Cyd, we have detected radioactivity in nucleotide fractions and in RNA and DNA. Degradation to nucleosides of the labeled triphosphate nucleotide fraction and of RNA showed that the radioactivity present was chromatographically identical to psi l Cyd. Half-saturation concentrations for the incorporation of [2-14C]psi l Cyd into the triphosphate nucleotide fraction and into RNA and DNA were 370, 280, and 94 microgram/ml, respectively, which were greater than 100-fold higher than those for tritiated cytidine. The incorporation of psi l Cyd was competitively inhibited by cytidine. Phosphorylation and incorporation of psi l Cyd into nucleic acids of P815 cells and of a P815 subline resistant to 1-beta-D-arabinofuranosylcytosine are about 2- to 20-fold higher than in P815 sublines resistant to psi l Cyd or to both 5-azacytidine and 1-beta-D-arabinofuranosylcytosine. These data suggest that the phosphorylation of psi l Cyd and possibly its incorporation into nucleic acids are essential for therapeutic activity in P815 leukemias. In vitro metabolic studies also suggest that psi l Cyd and 5-azacytidine are cross-resistant and that P815 cells resistant to psi l Cyd are collaterally sensitive to 1-beta-D-arabinofuranosylcytosine. These predictions were confirmed by therapeutic experiments carried out in mice bearing P815 leukemias.


Effects of iodine deficiency and high-fat diet on N-nitrosomethylurea-induced mammary cancers in rats. The effects of an altered content of dietary iodine and fat on the development of N-nitrosomethylurea-induced mammary tumors in rats were studied and correlated with thyroid and pituitary function studies. In three separate experiments, animals fed a semisynthetic diet containing 11.8% fat had an earlier time of tumor appearance and greater tumor burden than did controls maintained on a diet containing 4.6% fat. These diet-associated changes were markedly inhibited by ovariectomy, indicating that the tumor growth was hormone responsive. We examined the possibility that the diet with increased fat content enhanced tumor growth through alterations in prolactin metabolism but could find no consistent elevation in serum prolactin and no increase in pituitary prolactin synthesis in vitro. Our data further showed that rats on an iodine-deficient form of the high-fat diet had no greater tumor growth than did animals receiving an iodine-supplemented form of the same diet. We conclude from these results that iodine deficiency does not promote mammary tumorigenesis. An incidental finding of great interest was that ovariectomy led to a highly significant depression of thyroid-stimulating hormone production in vitro. This suggests that estrogens may directly influence thyroid-stimulating hormone synthesis in vivo and thus contribute to the sex-related differences in thyroid physiology.