Evidence for a morphological sex difference within the medial preoptic area of the rat brain. The present report demonstrates the existence of a marked sexual difference in the volume of an intensely staining cellular component of the medial preoptic nucleus (MPON) of the rat. Moreover, this sexual dimorphism is shown to be independent of several specific hormonal conditions in the adult, but significantly influenced, perhaps determined, by the perinatal hormone environment. Adult rats were gonadectomized and sacrificed 2 or 5-6 weeks later, or sacrificed after gonadectomy and priming with estradiol benzoate (2 microgram/day x 3) and 500 microgram progesterone, or testosterone propionate (TP, 500 microgram/day x 14), or the ingestion of propylthiouracil (0.15% of the diet) for one month, or following water deprivation for 24 h. These treatments did not affect the sexual dimorphism in the MPON and, in all groups, nuclear volume in the male animals was significantly greater than that of females whether nuclear volume was expressed in absolute terms or relative to brain weight. On the other hand, the volume of the MPON of the adult male castrated neonatally was significantly reduced when compared to that of the male castrated at the time of weaning, i.e. after the period of sexual differentiation of the brain. Consistent with the view that this nuclear region undergoes sexual differentiation is the fact that the volume of the MPON was significantly greater in female rats injected with 1 mg TP on day 4 of life than in oil-treated females. More subtle sex differences in the volume of the suprachiasmatic nucleus were also detected, as were several treatment effects. Although these differences may fall within the error of the analytical procedure, it is possible that hormone- or sex-dependent morphological differences exist elsewhere in the brain. Nevertheless, the gross sexual dimorphism in the MPON clearly demonstrates a possible morphological basis for the sexual differentiation of brain function.


Morphological changes in the cat cerebral cortex produced by superfusion of ouabain. The morphological changes following superfusion of the cat cerebral cortex with ouabain were studied. Autoradiography of [3H]ouabain was performed to study drug distribution. The resulting lesion consists of an upper vacuolated layer which is distinctly separated from an underlying region containing dark neurones. Ouabain is confined to the vacuolated layer. Swelling of apical dendrites and many presynaptic terminals are the main morphological changes occurring in the vacuolar layer. Depletion of synaptic vesicles, clustering of vesicles around the synaptic membrane, and the production of coated vesicles and cisternae are further changes found within presynaptic endings. Whilst swelling of fibrous astrocytes within the glialimitans occurs, this is not true of astrocytic processes elsewhere in ouabain exposed regions. Regions containing dark neurones are characterised by a general swelling of astroglial processes. The results strongly suggest that apical dendrites and presynaptic endings possess high activities of Na+, K+-ATPase whereas the activity on astroglial processes within the neuropil is relatively low. Astroglial swelling in areas of dark neurones is produced by some change in the chemical milieu surrounding the processes which appears unrelated to Na+, K+-ATPase inhibition.


Cat spinoreticular neurons: locations, responses and changes in responses during repetitive stimulation. Neurons in the lumbosacral spinal cord which project to the medial pontomedulary reticular formation were studied in the chloralose-anesthetized cat. Such neurons, identified by antidromic activation, were found predominantly in lamina VIII and medial lamina VII, and most were found to project to the contralateral reticular formation. Receptive fields for natural stimuli were generally complex, having various combinations of excitatory and inhibitory areas ipsi- and/or contralaterally. Adequate stimuli ranged from innocuous to noxious, with the stimuli required for decreasing a neuron's activity usually more intense than the stimulus required for increasing it. Electrical stimulation of hindlimb nerves indicated the presence of extensive convergence. Responses of spinoreticular neurons were found to decline during periods of repetitive stimulation. The response decrements were found to have many of the parametric features of behavioral habituation and were similar to response decrements previously observed in the medial pontomedullary reticular formation.


Studies on dopamine turnover in ovariectomized or hypophysectomized female rats. Effects of 17 beta-estradiol benzoate, ethynodioldiacetate and ovine prolactin. The effects of 17 beta-estradiol benzoate (EB), ethynodioldiacetate and ovine prolactin on dopamine (DA) turnover have been studied. As an index for a change in turnover, differences in DA depletion following tyrosine hydroxylase inhibition with alpha-methyl-tyrosine methylester were observed. DA was measured by means of mass fragmentographical analysis in the rat median eminence, the olfactory tubercle and the striatum. The actions of ethynodioldiacetate and ovine prolactin on DA turnover in various subregions of the rat median eminence were analysed by quantitative microfluorimetry. Repeated injections of EB to ovariectomized rats resulted in a significant increase of DA turnover in the median eminence. Administration of ethynodioldiacetate to ovariectomized rats almost significantly increased DA turnover in the olfactory tubercle. In the median eminence DA turnover was significantly increased only in the lateral palisade zone. In male hypophysectomized rats ovine prolactin increased DA turnover in both the lateral and the medial palisade zone of the median eminence. The results support the involvement of DA neurons in the control of prolactin and luteinizing hormone secretion. It is suggested that the tubero-infundibular DA neurons are involved in mediating the central inhibitory feedback actions of prolactin and gonadal steroids on prolactin and LH secretion, respectively.


Comparison of firing patterns in oxytocin- and vasopressin-releasing neurones during progressive dehydration. The electrical activity of neurosecretory cells in the supraoptic nucleus of the urethane-anaesthetized lactating rat was examined after periods of water deprivation ranging from 0-24 h. Supraoptic units were identified by antidromic activation following stimulation of the neurohypophysis, and classified as oxytocin or vasopressin cells according to their response during reflex milk ejection. In 65 vasopressin cells, dehydration increased the mean firing rate from 2.1 spikes/sec at 0 h to 6.8 spikes/sec at 24 h and caused a change from a slow irregular to a phasic firing pattern. Thus, after 6 h or more of dehydration, 84-100% of the vasopressin cells fired phasically, compared to 12% under normal conditions. In phasic vasopressin cells , the intraburst firing rates were closely related to the stages of dehydration, rising from a mean of 6.3 spikes/sec at 6 h to 12.0 spikes/sec at 24 h. However, no systematic relationship was observed between the stages of dehydration and the mean burst or silence durations. In 77 identified oxytocin units, dehydration increased the firing rate from 0.9 spikes/sec to 2.8 spikes/sec after 24 h, but only 3 (4%) of the cells showed phasic firing. Instead, the oxytocin units changed from a slow irregular to a fast continuous discharge. In conclusion, both vasopressin and oxytocin neurones are activated during chronic dehydration, but there is a marked difference in the pattern of their response. The phasic firing of the vasopressin cells may be important in increasing the occurrence of short interspike intervals and thus facilitating hormone release.


Subcellular mechanisms of lead neurotoxicity. The neurotoxic effects of inorganic lead (Pb) involve inhibition of calcium-dependent acetylcholine release and increases in calcium-dependent dopamine release. These apparently differential effects of Pb are associated with differing Pb-calcium (Ca) interactions: Pb blocks 45Ca binding to peripheral cholinergic ganglia and increases 45Ca binding to synaptosomes prepared from caudate nucleus (CN). Pb-induced increases in CN 45Ca binding did not result from nonspecific disruption of selective ion permeability of the membrane. Also, the Na-K ATPase-linked Ca extrusion system of synpatosomes was not affected by Pb. A Pb-sodium (Na) interaction was found such that elevation of intrasynaptosomal Na reversed effects of Pb on 45Ca binding. The intracellular localization of this effect appeared to be primarily at the mitochondrial level. Pb inhibited Na-induced release of 45Ca from preloaded mitochondria. This action may be translated into increased transmembrane flux of exogenous Ca, and thence into increased exocytotic events at the synapse. The apparently neurotransmitter-specific effects of Pb, cholinergic inhibition and dopaminergic augmentation, are hypothesized to result from different Pb-Ca interactions which are determined by the specific localization of Pb within nerve endings.


Angiotensin-induced thirst: effects of third ventricle obstruction and periventricular ablation. The role of periventricular tissue surrounding the anteroventral third ventricle (AV3V) in mediating drinking behavior induced by angiotensin II was investigated rats. Electrolytic lesions which bilaterally destroyed preoptic-hypothalamic periventricular tissue surrounding AV3V abolished drinking responses normally elicited by intracerebral injections of angiotensin. In another experiment, ventricular obstruction in AV3V had no effect on drinking of a palatable sweet milk solution while the drinking responses induced by peripheral versus central administration of angiotensin were dissociated. Drinking normally induced by lateral preoptic injections of angiotensin was no longer observed when AV3V obstruction prevented drug distribution via cerebrospinal fluid circulation to AV3V periventricular tissue; the drinking response induced by subcutaneous injection of angiotensin was enhanced, however, after placement of the ventricular obstruction. These results, coupled with the earlier observation that AV3V lesions also abolish drinking induced by subcutaneous angiotensin injectin, suggest that, after central or peripheral administration, angiotensin acts on AV3V periventricular tissue to arouse drinking. In contrast to centrally injected angiotensin, peripherally administered angiotensin does not contact receptors by entry and spread in cerebrospinal fluid. After peripheral injection angiotensin may contact sensitive AV3V tissue directly from blood, perhaps via the organum vasculosum of the lamina terminalis, a highly vascularized circumventricular organ within the AV3V which lacks a blood-brain barrier.


Aspects of influx and efflux of homovanillic acid of rat cerebrospinal fluid. In rats, different methods of perfusion (lumbar-cisternal or ventricular-cisternal) with artificial cerebrospinal fluid were performed at velocities of about 30 and 180 microliter/min. Levels of homovanillic acid (HVA) in the outflow were assayed with a semiautomated fluorimetric technique. Intravenous administration of 20 microgram HVA did not substantially enhance the outflow of this acid, indicating that in our preparations HVA found in the perfusate is of central origin. In the lumbar-cisternal preparation probenecid (200 mg/kg, i.p.) was found to inhibit the efflux of a significant fraction of HVA added to the medium, at both perfusion rates. The proportions of HVA eliminated by a probenecid sensitive transport was much higher at the lower rate of perfusion. Following probenecid the increase of endogenous HVA in the ventricular-cisternal perfusate was higher at a lower rate of perfusion. We determined the turnover rate of HVA in the whole brain and compared this value with the HVA outflow in the various preparations. The highest efflux of HVA was found in the ventricular-cisternal preparation during probenecid treatment and did not appear to be dependent upon the rate of perfusion. A maximal value of 3.5% of HVA formed in the central nervous system was found to be released into the cerebrospinal fluid.


Amphetamine-induced increase in rat cerebral blood flow: apparent lack of catecholamine involvement. The influence of D,L-amphetamine (5 mg/kg i.p.) on regional cerebral blood flow (CBF) in rats has been studied after surgically or pharmacologically induced depletion of brain catecholamines. (1) Bilateral removal of the superior cervical ganglion (one week before the experiment) did not prevent the amphetamine-induced augmentation of CBF present in intact animals to 2--4 times above the control value. Maximal changes occurred in the frontal and parietal cortex. (2) Destruction of the ascending noradrenergic pathways by uni- or bilateral injections of 6-hydroxydopamine, which decreased the noradrenaline (NA) level in the frontal cortex by 89%, was ineffective in abolishing the increase in CBF caused by the drug in the frontal cortex. (3) The involvement of other catecholaminergic systems was excluded by pretreatment of the rats with reserpine plus a-methyl-p-tyrosine which reduced the levels of NA, dopamine and adrenaline in the frontal cortex with 92, 97 and 99% respectively. Such treatment did not alter the effect of amphetamine on CBF in the frontal cortex. The results support the hypothesis that the action of amphetamine on CBF is not mainly mediated by catecholamines.


Postural and kinetic coordination following cortical stimuli which induce flexion movements in the cat's limbs. The relationship between movement and the postural adjustment associated with it has been studied following flexion movements of a limb induced by cortical stimulation in chronically implanted cats. Variations in the vertical force exerted by each limb, as well as the displacement of the centre of gravity (projected onto the plane of stance), were investigated. It was established that: The pattern of postural adjustment observed in these conditions is diagonal; that is to say, flexion of one limb is accompanied by diminution in the vertical force exerted by the diagonally opposite limb and an increase in the force exerted by the other two. The displacement of the projected centre of gravity is sterotyped during the first phase of variation in forces. Its extent may be considered as an index of the efficiency of postural adjustment. The study of the latencies of the changes in force indicates, at least in the case of hindlimb movements, that postural adjustment is an active phenomenon and does not arise passively from the interplay of purely mechanical forces. In the same conditions, an analysis of the time course and latency of recorded changes in force indicates that postural adjustment is not triggered reflexly by the disequilibrium brought about by movement but is a result of a central nervous command.


Influence of pyramidotomy on limb flexion movements induced by cortical stimulation and on associated postural adjustment in the cat. Flexion movements induced by cortical stimulation and the associated postural adjustments in bilaterally pyramidotomised cats have been studied by means of an apparatus which measures separately the changes of force under each limb in the upright position of the animal. The results show: (1) The general motor behaviour of the animal is not affected by the lesion. The principal deficit is loss of tactile placing reaction in the forelimbs; there also appears a state akin to a vestibular syndrome when a light restraining harness is placed around the back and chest. (2) Stimulation of the motor cortex continues to evoke flexion movements of the contralateral limbs together with associated postural adjustment. Coordination between movement and postural adjustment is generally similar to that observed before pyramidal section, and response thresholds are unchanged. (3) Measurements reveal great quantitative modification both of the movement and the postural adjustment after the lesion. Weight shift latencies are increased and more dispersed in time, while the weight shifts themselves are reduced in amplitude and speed. (4) All these changes are more marked in the case of forelimb, rather than hindlimb, flexion, emphasising the greater degree of pyramidal influence on forelimb activity in the normal animal. (5) The results as a whole underline the ability of non-pyramidal systems, under the control of the motor cortex, to bring about both limb flexion and the associated postural adjustments in the absence of the corticospinal pathway.


Single unit activity in cat prefrontal and posterior association cortex during performance of spatial reversal tasks . Relationships between the performance of complex learning tasks and the firing patterns of single unites in prefrontal and posterior association areas of the neocortex were studied in 9 cats. Recording data from 2 additional cats served as controls for sensory and motor aspects of the tasks employed. During recordings, animals sat in a box, their heads fixed to a stereotaxic instrument, and performed a non-sensory spatial reversal or delayed-alternation task by pressing 1 of 2 lateralized retractable levers. Units obtained were classified into 5 basic types according to their correlation with aspects of the tasks. The first 3 types were related directly to sensory, motor and motivational aspects of the tasks. The fourth type was named associative, as it contained units which changed firing rates in advance or as a consequence of environmental events, rather than in direct relation. Discharge changes of units of the fifth type were rated as non-specific or non-specifiable. Between recording areas and tasks a striking similarity of unit firing patterns was obtained. Differences were observed only between the reactivity of polymodal cells in prefrontal and in posterior fields, and in the proportions of units revealing mnemonic and motor response aspects. It was concluded that a considerable functional overlap between prefrontal and posterior association areas of the cat with respect to processing of complex learning tasks exists on the single neuron level.


Consequences of blocking the nerve with a local anaesthetic on the evolution of multiinnervation at the regenerating neuromuscular junction of the rat. A transient multiinnervation is observed in adult rats during reinnervation of the soleus muscle. Chronic blockage of the sciatic nerve with a cuff of the local anaesthetic lidocaine during reinnervation increases the number of multiply innervated muscle fibres. This result indicates that the selective elimination of redundant nerve terminals is regulated by the activity of the innervated muscle.


Substance P in the amygdaloid complex, bed nucleus and stria terminalis of the rat brain. Radioimmunoassay and immunohistochemical techniques have demonstrated the presence of substance P in the medial and central nuclei of the amygdala and the bed nucleus of the stria terminalis. Hemisections and micro-knife cuts transecting the anterior, posterior, medial, lateral and ventral connections to the amygdala did not modify the content of substance P in the amygdala. In addition knife cuts totally isolating the medial amygdaloid nucleus from lateral and anterior-posterior connections did not reduce the substance P content of the medial nucleus, but produced a 70% reduction in the substance P content of the central nucleus. These results suggest that substance P containing neurones in the medial and central amygdaloid nuclei do not receive substance P projections originating outside the amygdala. However, there appears to be a short substance P projection from the medial nucleus to the central nucleus.


Intermittent mandatory ventilation and controlled mechanical ventrilation without positive end-expiratory pressure following cardio-pulmonary bypass. In a group of 18 male patients undergoing coronary artery bypass grafting with cardiopulmonary bypass, the overall incidence of post-operative atelectasis was 60%. Nearly three-quarters occurred during anaesthesia. After operation there was no difference whether CMV or IMV without PEEP was provided overnight. Atelectasis already present did not improve and further atelectasis occurred. A role for IMV is not excluded, since it facilitates the use of PEEP. Many factors operate and interact to provoke atelectasis during anaesthesia, which increases post-operative morbidity. Many of these factors are prevertible or reversible if their physiological basis is understood. Optimal post-operative ventilation should be tailored to the needs of the individual patient and demands close co-operation between anaesthetist and surgeon.


The mechanism of anaesthesia: a new hypothesis based on the effects on electrical properties of a model membrane: preliminary studies. Changes induced by inhalational anaesthetic agents were investigated in a simple membrane model. All the agents tested increased the membrane's electrical capacitance and conductance linearly, proportional to concentration and potency, and decreased its dialectric strength independently of concentration. When protein was substituted for lipid, the same agents had no significant effect on capacitance, conductance, or dielectric strength. Based on these findings, the following hypothesis is presented. Anaesthetic agents exert their influence on the lipid portion of nerve membranes. They enhance excitability by reducing the dielectric strength of the membrane and depress excitability by increasing the membrane's dielectric constant and passive conductance. The degree of excitability or narcosis at any instant depends upon the relative magnitude of these opposing effects.


The effect of ethanol on the ventilatory responses mediated by the peripheral chemoreceptors in man. The ventilatory responses mediated by the central and peripheral chemoreceptors were separately assessed in eight healthy volunteers before and after the oral ingestion of ethanol in a dose of 0.75 ml/kg. No significant depression of the central response was observed, but a significant depression of the peripheral response was observed at 25 and 95 minutes after the consumption of ethanol. The peripheral chemoreceptor stimulus was the simultaneous increase of hypoxia and hypercapnia and this novel method is described.


[Critical study of the hemodynamic status during anesthesia]. Physicians must choose the anaesthetics for their patients and select the methods to cheek their haemodynamic status. Experimental works do not always bring sufficient information to help them in their daily practice. Circulatory reaction to a pharmacodynamic agent is diffuse and non-specific. Some examples drawn from theoretical considerations and practical situations support this view. The difficulty of assessing the actual damage caused by one apparent variation still remains. On the other hand, haemodynamic indices such as PA, CVP, dP/dt are composed of elementary data that are interdependent; so indices are interdependent, too. A puzzling fact is that depressing anaesthetics can initiate severe haemodynamic crisis, generally badly tolerated by tissues, but better tolerated by myocardium which is not definietely altered if the anoxia inflicted is accompanied by a real decrease in cardiac work. This explains the often slight effect of short anesthetic overdosage. Previous haemodynamic variations to be countered relative to anaesthesia are not really known. Biochemistry of the venous coronary blood does not yet bring striking features in this field.


An evaluation of the Bentley PFF-100 transfusion filter. The effects of a new transfusion filter (Bentley PFF-100) on stored whole blood have been examined. Six filters were preloaded by passage of two units of outdated type-specific bank blood and the effects of filtration on a third unit of 21-day-old blood flowing under 19.95 kPa (150 mm Hg) pressure were measured. Filtration did not significantly alter erythrocyte or leukocyte count, total or plasma haemoglobin, red cell fragility, plasma sodium, potassium, albumin, or globulin. Platelet counts were reduced by 31%. Removal of microaggregates, assessed by Coulter counting, screen filtration pressure, total screen protein, wet and dry weight of material retained and scanning electron microscopy, was shown to be effective over the entire range of particle size. In comparison with other transfusion filters previously examined in this laboratory, the Bentley PFF-100 filter combines the feature of moderately efficient microaggregate removal with rapid blood flow rate. Compared to its predecessor, the Bentley PF-127, this new filter design has improved flow characteristics without loss of efficiency of microaggregate removal.


Comparison of two benzodiazepines for anaesthesia induction: midazolam and diazepam. Ro 21-3981 is a newly synthesized water soluble benzodiazepine derivative. Its pharmacological properties are similar to diazepam. This investigation was designed to establish the effective induction dosage of Ro 21-3981 and to compare it with diazepam for induction of anaesthesia. The ED50 for Ro 21-3981 induction is 0.15 mg/kg and ED 100 is 0.2 mg/kg. Ro 21-3981 is one and one-half times as potent as diazepam (0.3 mg/kg) and more rapid in action. There is significantly less pain on injection with Ro 21-3981 as compared to diazepam. Cardiovascular stability and apnoea were observed with both drugs. Ro 21-3981 is a promising anaesthetic induction drug that merits further human study.


The uptake and excretion of nitrous oxide in the newborn. In the newborn, the uptake of nitrous oxide into the alveolus is very rapid, as is the fall in alveolar levels following withdrawal of the agent. During recovery relatively large volumes of nitrous oxide are excreted in the first minute, after which the excretion rate rapidly declines, most of the gas having been eliminated.


Management of chronic dorsal root pain with epidural steroid. Twenty-eight patients presenting with typical dorsal root pain of more than six months' duration were given a series of three epidural injections of methylprednisolone acetate. The patients were grouped according to known aetiologic factors. Improvement in symptoms was most marked in the post-herpetic group where 86% were free of pain after six months. In the post-trauma/surgical group, 50% of patients were free of pain after six months, while in the idiopathic group only minimal improvement occurred. The side-effects of steroid administration into the epidural space were in all cases minor.


Caudal anaesthesia in paediatric patients. We have described our technique of caudal anaesthesia in children. This technique was used for relief of post-operative pain in 120 patients. Relief of pain was complete. Post-operative recovery was quiet and comfortable, with no side effects. The surgeons, the parents and recovery room personnel were satisfied. There were no surgical or anaesthetic complications.


Anaesthetic considerations in caesarean section for quadruplets. A case of a caesarean delivery with epidural analgesia of a term parturient with quadruplets is presented. Maternal considerations of hypotension, respiratory embarrassment and aspiration of gastric content and foetal considerations of prematurity and impaired placental function are discussed relative to the use of general anaesthesia or epidural analgesia.


Accidental over-dose of alfathesin under general anaesthesia: case report. Under general anaesthesia, 35 ml of alfathesin was accidentally injected intravenously. There was no change in the pulse rate and blood pressure. However, two hours later, at the end of the operation, the rectal temperature was 31 degrees C. There were no long-lasting effects of the overdose.


Liquid nitrogen storage and human chromosome composition of human-mouse hybrid cells. The human chromosome complement of eight human-mouse hybrid cell strains was studied with fluorescent banding before and after rejuvenation of these cells from liquid nitrogen storage. The karyotype remained essentially stable in several cell strains, while in others both human and mouse chromosomes were lost. A third group demonstrated loss of only human chromosomes. The loss of particular human chromosomes after the rejuvenation process was shown to be statistically significant in four of the eight hybrid cell strains. These preliminary results suggest that karyotypic constancy cannot be assumed in human-mouse hybrid cells subjected to rejuvenation from liquid nitrogen storage.


Cytological effects of microwave radiation in Chinese hamster cells in vitro. Cytological effects were investigated in the Chinese hamster cell line (CHO-K1) exposed to microwave radiation of 2450 MHz frequency and incident power of 25 to 200 W for a period of 30 min. Nuclear vacuoles, pycnotic and decondensed chromosomes were observed in cells exposed to 25 W under elevated temperature conditions (uncontrolled temp). In addition a significant increase in chromosomal breakages/cell was observed. Cells exposed to relatively higher power, 75-200 W, under hypothermic conditions (29 degrees C) revealed no significant increase in either nuclear vacuoles or other chromosomal anomalies over control cells. Radiation-induced temperature elevation appears to be an essential factor in the cytological effects of microwave.


Hyperthermia and host-cell reactivation of adenovirus 12. Exposure of cultured human fibroblasts to hyperthermia delayed the host-cell reactivation of UV-irradiated human adenovirus type 12 (AD12). The experimental design consisted of irradiating human AD12 with UV doses ranging from 180 to 1800 ergs/mm2, infecting human cell populations at 37 degrees C, exposing the infected cells for 7 h to 39.5 degrees C and 41.8 degrees C, returning them to 37 degrees C and estimating the frequency of cells with intranuclear viral inclusion bodies (IB) 41 and 89 h after hyperthermia treatment. Hyperthermia reduced the fractions of fibroblasts with viral IB in the 41 h samples. By 89 h the capacity to form IB in the treated cells was comparable to that in control cells. Hyperthermia of 39.5 and 41.8 degrees C for 7 h did not affect the replication of nonirradiated AD12. The pattern of host-cell reactivation of AD12 following hyperthermia was compared to that in DNA repair deficient xeroderma pigmentosum cell populations.


Chinese hamster cell lines resistant to the cytotoxic action of fluoride. The proliferation and efficiency of colony formation of a Chinese hamster ovary cell line, CHO, was found to be inhibited by concentrations of fluoride greater than or equal to 10(-3) M. From mutagenized populations of CHO cells, clones were isolated that were from 1.6 to 13 times more resistant than the wild-type to the cytotoxic action of fluoride. The resistant clones were found to be stable in the absence of selection. The fluoride sensitivity of wild-type and fluoride resistant clones was not altered by changes in the pyruvate concentration in the culture medium, indicating that the cytotoxic effect of fluoride is not due to the action of fluoride on the glycolytic pathway. On the other hand, both the incorporation of 3H-leucine into acid precipitable material and the distribution of polyribosomes were sensitive only to fluoride concentrations that were cytotoxic, suggesting that the molecular basis of fluoride induced cytotoxicity in both wild-type and fluoride resistant cells is the sensitivity of protein synthesis to fluoride. At concentrations of fluoride at which the wild-type cells are inhibited but fluoride resistant cells are not, the intracellular concentration of fluoride in the fluoride resistant cells was found to be 1/5 to 1/10 that of the wild-type, suggesting that fluoride exclusion is the basis for resistance in the resistant lines.


Effects of interactions between different culture fractions of 'phosphobacteria' and Rhizobium on mycorrhizal infection, growth, and nodulation of Medicago sativa. Interactions between cell-free culture supernatants, cells, and the whole cultures of Rhizobium and phosphobacteria with endomycorrhizal fungi and their effects on growth and nutrition of Medicago sativa grown in a low-phosphate soil were studied. A satisfactory nodulation was greatly dependent on the mycorrhizal symbiosis. Cell-free supernatants of Rhizobium and phosphobacteria improved plant growth, nodulation and mycorrhiza formation. The activity of phosphobacterial culture seemed to be due mainly to the supernatant and the possibility of plant hormones contained in this culture fraction being involved in such interactions is discussed. An increase of the overall pool of soluble P in soil by the inoculated phosphobacteria cells was not found in the conditions of this experiment. It was noteworthy that the best positive effect was achieved by the treatment which consisted of the whole cultures of Rhizobium, phosphobacteria, and the mycorrhizal fungi applied all together.


Pseudomonas maltophilia: identification of the hydrocarbons, glycerides, and glycolipoproteins of cellular lipids. The nature of quantity of the free lipids of Pseudomonas maltophilia have been examined. Lipid components identified as phosphatidyl ethanolamine, phosphatidyl glycerol, diphosphatidyl glycerol, phosphatidylamino acid derivatives and phosphatidic acid accounted for about one-half of the total lipids and are generally typical of gram-negative bacteria. The remaining lipids were identified as a complex family of aliphatic hydrocarbon isomers (C22--C32) and a mixture of glycolipoproteins. These components are generally atypical lipids of bacteria. The glycolipoproteins were composed of a broad range of amino acids, ketodeoxyotulonic acid, mannose, glucose, 4-aminopentose, glucosamine, phosphates, and a relatively large quantity of fatty acids, principally OH-13:0. The total esterified fatty acids consist of cyclopropane, iso- and anteiso-branched and normal saturated and unsaturated chains in the range from C14 to C20. Amide bound fatty acids consist of OH 11:0, OH-12:0, OH-13:0, and methyl-branched OH-13 in addition to methyl-branched and normal saturated and unsaturated chains.


Presence and activity of the hexosemonophosphate shunt in a marine pennate chemoorganotrophic diatom, Nitzschia alba, clone Link 001. A commonly occurrring chemoorganotrophic diatom, Nitzschia alba, clone Link 001, stores primarily fatty acids and utilizes the hexose monophosphate shunt as a major pathway in hexose oxidation. The presence and activity of this pathway in exponentially growing N. alba was demonstrated by the growth on potassium gluconate as a sole carbon source, in vitro assay of 6-phosphogluconate dehydrogenase (EC 1.1.1.44), and in vivo radiorespirometric evaluation using [14C]-glucose or -gluconate. Radiorespirometry demonstrated that the hexose monophosphate and Embden--Meyerhoff--Parnas pathways account for 46 and 54% of hexose oxidation, respectively. The predominance of fatty acid storage plus the probable utilization of some C4--C5 hexose monophosphate intermediates in biosynthetic activities support the relatively high hexose monophosphate activity in these exponentially growing cells. Radiorespirometric values are supported by the absence of Enter-Doudoroff activity as determined by the lace of 2-keto-3-deoxy-6-phosphogluconate aldolase (EC 4.12.14) and 6-phosphogluconate dehydratase (EC 4.2.1.14) activity.


[Actinomycetes antagonistic to fungi and not affecting Rhizobium meliloti]. The effects of 481 actinomycetes isolated from agricultural soils supporting good growth of alfalfa or clover on two efficient strains of Rhizobium meliloti A2 and S14 were studied. Strain A2 was inhibited by 28% of the isolates and strain S14 was inhibited by 31% of them. No significant difference was found between the resistance of both actinomycete strains. The effects of the 288 isolates not affecting R. meliloti on six fungi were also studied. The most sensitive fungus was Stemphylium sarcinaeforme inhibited by 20% of the isolates, while Fusarium culmorum was the most resistant fungus and was inhibited by only 6% of the isolates. Thirteen isolates inhibited four to six fungi. In an autoclaved greenhouse soil, isolate 181 which inhibited the six fungi tested significantly reduced the population of the phytopathogenic fungus F. oxysporum f. sp. medicaginis and eliminated the inhibitory effect showed by this fungus on strain A2 of R. meliloti.


Surcrose uptake by Neisseria denitrificans. Neisseria dentrificans possesses an inducible sucrose-uptake system. Sucrose is transported as the free sugar and is rapidly converted into polymerized material. Uptake is inhibited by glucose, cyanide, uncouplers of oxidative phosphorylation, and sulfydryl reagents. Glutamate serves as an energy source for uptake. The importance of this phenomenon in the oral environment is discussed.


Visual micromethod for assay of fungal growth. A visual micromethod for measuring antifungal effects on germination and growth is described. The antifungal agent griseofulvin and the fungus Trichophyton mentagrophytes were used as materials to compare the micromethod with a standard assay based on dry mycelial weight. The micromethod was more sensitive than the weight method in detecting the minimum inhibitory concentration of griseofulvin (0.18 and 35 microgram/ml, respectively). At higher concentrations of griseofulvin (22.5 microgram/ml), the micromethod measured minimal fungal growth that was undetectable on a weight basis. The micromethod showed that griseofulvin does not change the number of spores forming germ tubes. Progressively severe alterations in fungal morphology occured as the concentration of griseofulvin was increased from 0.09 to 22.5 microgram/ml.


Effect of respiratory deficiency and temperature on the mitochondrial lipid metabolism of Aspergillus niger. A comparative study of the mitochondrial lipid composition of a wild strain (V 35) and a respiratory-deficient mutant (rd3) of Aspergillus niger grown at different temperatures (25, 30, 35, and 40 degrees C) has been performed. The lipid spectrum, though qualitatively similar, differs quantitatively in both cases. At the optimum growth temperature (30 degrees C) depletion in ergosterol (40%) and cardiolipin (52%) was observed. This probably indicates the formation of defective mitochondria in the mutant with a resultant impaired respiratory system. Complete depletion of cardiolipin species containing fatty acid (20:5omega3) in the respiratory-deficient mutant suggests a possible role of this lipid in mitochondriogenesis at least in A. niger. The effect of temperature is predominantly on the degree of unsaturation and sterol ester formation. The linoleic acid (18:2omega6) content decreases with a concomitant increase in oleic acid (18:1omega9) content as the growth temperature increases for both cell types. Some morphological changes and effects on the vegetative life cycle have been observed with variation in growth temperature in the wild type and also in the mutant form.


[Morphological, physiological and taxonomic studies of Bacillus azotoformans]. Seventeen strains of the new species Bacillus azotoformans were isolated by enrichment culture in peptone broth inoculated with pasteurized soil and then incubated under N2O at 32 degrees C. The bacterium is a Gram-negative rod, motile with peritrichous flagella, which produces oval spores without exosporia in swollen sporangia. However, the cells have thick walls, mesosomes, and persistent septa characteristic of Gram-positive bacteria. The bacterium lacks fermentative activity, does not attack carbohydrates, has complex growth requirements, and will grow anaerobically only if one of the following electron acceptors is present: NO3-, NO2-, N2O, S4O6--, or fumarate. Nitrate, nitrite, and nitrous oxide are denitrified with the production of N2. The microorganism is mesophilic, gives a positive oxidase reaction, synthesizes a type c cytochrome, and does not hydrolyse gelatin, starch, or "Tween 80." Poly-beta-hydroxybutyric acid is snythesized when the bacterium is grown in a medium containing DL-3-hydroxybutyrate. The following enzymes are present: nitrate reductase A, respiratory nitrite reductase, tetrathionate and fumarate reductases, and L-glutamate dehydrogenase. The following enzymes are absent: thiosulfate reductase, urease, lecithinase, arginine dihydrolase, phenylalanine deaminase, and catalase. For the 17 strains, the mean value of the G = C percent of the DNA is 39.8 +/- 1.2. All the strains are highly similar.


Glucose metabolism in Pycnoporus cinnabarinus. It was demonstrated by the measurement of enzyme activities and by radiorespirometric assays that the basidiomycete Pycnoporus cinnabarinus metabolizes glucose through the glycolytic pathway and the pentose phosphate cycle.


Improved production of citric acid by a diploid strain of Aspergillus niger. Aspergillus niger strain CGU 87 was treated with UV radiation and some auxotrophic mutants were obtained. These mutants were less productive than CGU 87, which produced an average of 7.4% citric acid. All possible crosses in pair wise combinations were carried out between these auxotrophs, and three heterokaryons were synthesised. Finally, one heterozygous diploid was isolated from each of them. These heterokaryons and diploids showed improved productivity when compared with their component parents, but except in one diploid D5, all others produced less citric acid than CGU 87. The yield of D5 exceeded that of CGU 87 by 1.2 times and it produced 9% citric acid. This is a significant improvement and the increased productivity seems to be the result of successful adaptation of D5 to its fermentation environment.


Treatment of psoriasis with orally administered 8-methoxypsoralen and long-wavelength ultraviolet radiation. Between May 1976 and September 1977, 51 patients with severe psoriasis were treated with orally administered 8-methoxypsoralen followed by exposure to high-intensity long-wavelength ultraviolet radiation. Clearing of psoriasis occurred in 40 patients (78%) and marked improvement in 5 (10%). Of the remaining patients three (6%), who had generalized erythroderma, failed to respond to this therapy. The mean number of treatments required for clearing was 37.5. No serious side effects were noted clinically, by ophthalmologic examination or by laboratory testing. This therapy has some advantages over conventional types of treatment now used for severe psoriasis, but also has limitations. It appears to be an effective method of treatment for ambulatory patients. Further long-term follow-up studies are required to evaluate its side effects.


Relation of amniotic fluid lecithin/sphingomyelin ratio and fetal asphyxia to respiratory distress syndrome in premature infants. In a group of 81 prematurely delivered infants the amniotic fluid lecithin/sphingomyelin ratio was related to functional fetal lung maturity. Intrapartum fetal asphyxia occurred in 33% of the group. An association between fetal asphyxia and the development of respiratory distress syndrome (RDS) in the infant was observed. When pulmonary maturity was borderline according to the amniotic fluid assessment, intrapartum fetal asphyxia was associated with an increased risk for development of RDS in the infant.


Use of a topical antibiotic spray in vaginal surgery. During vaginal hysterectomy with or without colporrhaphy a topical aerosol spray containing neomycin sulfate, polymyxin B sulfate and zinc bacitracin was used in 50 patients to decrease the change of postoperative pelvic infection; a placebo spray was used in another 50 patients. All patients were treated preoperatively with povidone iodine and postoperatively with nitrofurantoin and an antibacterial irrigating solution for the bladder if catheter drainage was necessary. The frequency of postoperative pelvic infection was 16% in the group sprayed with the antibiotic combination and 34% in the group sprayed with the placebo, a significant difference (P less than 0.05).


Cardiac conduction abnormalities and Stokes-Adams attacks in myotonic dystrophy. Myotonic dystrophy is a well known cause of cardiomyopathy. While various cardiac conduction abnormalities have been described in patients with myotonic dystrophy, so far only sporadic cases of Stokes-Adams attacks have been reported. Of 27 patients with this disease various conduction disturbances were detected in 17 (63%), 5 of whom presented with Stokes-Adams attacks and were found to have intracardiac conduction defects. The prognosis in four of the five patients was greatly improved with permanent pacemaker implantation.


Adverse biochemical and clinical consequences of furosemide administration. Nurse monitors collected clinical and laboratory data from 204 hospitalized patients receiving furosemide (122 men and 82 women; mean age 69.6 years). Biochemical abnormalities and clinical problems definitely or probably induced by any drug occurred in 70.6% and 49.0% respectively of the patients, and were attributed to furosemide in 81.3% and 13.0% respectively of these patients. The most important clinical events were dehydration and hypotension. Furosemide-induced hypochloremia, hypokalemia and hyponatremia occurred in 35.8%, 25.0% and 24.5% of the patients respectively. Most of the biochemical changes were slight, and only 3.9% of the patients had a furosemide-induced decrease in the serum potassium concentration to less than 3.0 mmol/L. Surprisingly, 24.5% of the patients also manifested drug-induced hyperkalemia. Administration potassium supplements or spironolactone, or both, concurrently with furosemide was responsible in most cases for the development of hyperkalemia. The occurrence of drug-induced adverse effects after 2 weeks of hospitalization was significantly associated (P less than 0.05) with subsequent prolongation of hospitalization. The high frequency of drug-induced events warrants careful monitoring of all patients receiving furosemide in spite of the low frequency of serious toxic effects produced by the drug.


Failure of diagnosis as a factor in tuberculosis mortality. In British Columbia between January 1970 and December 1974 active tuberculosis was diagnosed only after death in 69 cases; this was more frequent for miliary tuberculosis (31% of reported cases) than for advanced pulmonary tuberculosis (3% of cases). Although 28% of the patients were more than 75 years old, some were much younger, and 38% of the latter were alcoholics. More than 50% of the patients had been hospitalized before death, for a mean of 14.5 days; they were most frequently thought to have pneumonia or cancer at the time of death. Clearly, increased awareness of the continuing presence of tuberculosis in our society is needed.


Septicemia in children with leukemia.

Septicemia in children with leukemia. A review of the hospital records of 164 children with leukemia diagnosed between January 1969 and December 1975 disclosed 51 episodes of septicemia in 43 patients; 57 infectious agents were isolated. Gram-positive bacteria were isolated as frequently as gram-negative bacteria, each type accounting for 45.6% of all the agents isolated. Only 2 of the 24 episodes of exclusively gram-positive septicemia were fatal, whereas 9 of the 23 episodes of exclusively gram-negative septicemia were fatal. The mean duration of neutropenia was 5.6 days in patients with gram-positive septicemia and 19.5 days in patients with gram-negative septicemia, a significant difference (P less than 0.01). Gram-positive septicemia was diagnosed after a mean of 5.9 days of hospitalization and gram-negative septicemia after a mean of 29.0 days, also a significant difference (P less than 0.001). In this exclusively pediatric population of leukemic patients gram-positive agents have to be considered as potential pathogens, and initial antibiotic therapy must be selected with this fact in mind.


Surveillance of the swine influenza vaccination program at the Royal Military College, Kingston. In a prospective study symptoms appearing in a previously healthy population within 6 weeks after inoculation with monovalent swine influenza vaccine (A/New Jersey/76) were tabulated. Of the 703 persons (ranging in age from 17 to 55 years) participating in the follow-up 54% reported experiencing symptoms, usually within 24 hours of vaccination; the symptoms were usually minor and none of the participants displayed evidence of Guillain-Barré syndrome.


The impact of a new geriatric program in a hospital for the chronically ill. Reports of the rapidly increasing proportion of persons aged 65 years and more in Canada and the resultant need for changes in the country's health care system prompted experimental changes in the operation and training procedures at St. Mary's of the Lake Hospital, Kingston, Ont. Aimed at better patient care and at better education of medical house staff in geriatrics and long-term care, the revised program is permeated with the philosophy of rehabilitation. It includes full-time staff, a geriatric outpatient clinic, a day hospital, a team approach to patient care (with regular team audits), problem-oriented medical records, a formal physical medicine section with a district inpatient unit, and an intensive inservice education program. After the first year of the program patient outcome had improved and more efficient use was being made of continuing care beds because of larger numbers of patinets being discharged home after shorter stays. This may be one avenue for deceleration of our country's dismal rate of institutionalization.


Effectiveness of neonatal transport.

Effectiveness of neonatal transport. The condition of 259 infants transferred to the neonatal intensive care unit (NICU) of the Montreal Children's Hospital from Oct. 1, 1974 to Mar. 31, 1975 was evaluated. Their transport was provided by personnel and equipment from the Montreal Children's Hospital. When the transport team arrived at the referring hospital hypothermia (temperature of less than 36 degrees C) was present in 25.2% of the 163 infants for whom complete temperature measurements were available. Most (77.3%) of the infants were warmed during transport and only 3.1% arrived at the NICU with a temperature of less than 35 degrees C. The mortality was significantly higher in babies of all birth weight groups whose core temperature had been below the optimal temperature for survival (36 to 37 degrees C). It appears that the use of appropriate equipment and trained personnel can reduce the incidence of hypothermia and therefore the mortality in infants requiring transfer.


Fetal exsanguination associated with antepartum hemorrhage. Fetal bleeding in utero is infrequent. It is usually life-threatening but can be treated successfully in most cases if recognized early. Four cases are described and it is suggested that screening for fetal blood be done in all instances of antepartum hemorrhage.


Evaluation of three antibiotic programs in newborn infants. Emergence of gram-negative bacteria resistnant to a number of antibiotics in intensive care nurseries for neonates emphasizes the need for alternatives in antibiotic combinations. One commonly used combination, gentamicin-ampicillin, and two newer combinations, tobramycin-cephalothin and amikacinampicillin, were evaluated prospectively in 60 newborns in such a nursery. Subjects were randomly assigned to one of the above therapy groups. Dosages in mg/kg.d were 100 for ampicillin and cephalothin, 6 for gentamicin and tobramycin and 15 for amikacin. Aminoglycoside serum concentrations, clinical tolerance and toxicity were monitored. Aminoglycoside concentrations after intravenous administration of the drugs were within the expected range (gentamicin and tobramycin 4 to 6 microgram/mL and amikacin 15 to 20 microgram/mL). There was no hematologic, renal or hepatic toxicity attributable to antibiotic therapy and the combinations were tolerated equally; no bilirubin displacement was detected in vitro or in vivo.


Adriamycin in combination for the treatment of breast cancer: a Southwest Oncology Group study. Patients with advanced breast cancer who had not previously received chemotherapy were treated on a three-arm prospective study: adriamycin day 1 plus 5-FU on day 1 and 8 (AF), adriamycin day 1, plus 5-FU day 1 and 8, and cyclophosphamide day 1 (AFC), and adriamycin day 1 plus 5-FU day 1 and 8, cyclophosphamide day 1 and methotrexate day 1 (AFCM). These courses were repeated every 21 days. The response rate was 44/105 (42%) AF, 44/103 (43%) AFC and 52/105 (49%) AFCM. The length of response was 22, 33 and 35 weeks, respectively, for AF, AFC and AFCM (P = 0.21). The median survival, 64 weeks, was equal in all three limbs. The major toxicity was leukopenia. Twenty-eight percent developed a WBC of less than 2,000/microliter, which resulted in seven deaths (2.2%).


Surgical adjuvant chemotherapy: results with one short course with cyclophosphamide after mastectomy for breast cancer. One single six-day course with cyclophosphamide (total dose 30 mg/kg) was given immediately after mastectomy to 507 breast cancer patients, with 519 randomized controls receiving no adjuvant chemotherapy. The control group now has 234 recurrences and 196 deaths, and the treatment group 175 recurrences and 146 deaths. The differences of 59 recurrences and 50 deaths in favour of the treatment group are significant with p values less than 0.001 and less than 0.01 respectively. The differences in recurrence rates increased gradually, reached 10.71% four years after mastectomy (p less than 0.001), and remained at the same level for another 6 years. The differences in death rates increased until 6 years after mastectomy, and was 10.48% after 10 years. With this pattern, the mechanism is probably not a delay in onset of clinical recurrences, but a definite reduction of recurrence rates due to tumoricidal chemotherapy. Prognostic factors or menstrual state had apparently no influence on the effect of this type of adjuvant chemotherapy. Side effects were of short duration and very moderate. Since there was a good effect in the prognostically most favourable groups of patients, treatment of such cases seems therefore also justified. The same chemotherapy course given 3 weeks after mastectomy seemed without effect.


A correlation of serum C3DP levels with chemotherapeutic management of cancer patients. Serial serum C3DP levels of 33 patients at our institution have been followed for up to 10 months. Individuals experiencing periods free of symptoms and signs of proliferating or expanding malignant disease, had C3DP levels which remained below 150 microgram/ml. Patients with active or recurrent disease, while on chemotherapy, had elevated C3DP values (greater than 150 microgram/ml). Serum C3DP values declined abruptly following treatment which resulted in major reduction of tumor mass. Decreases in C3DP levels from values above 150 microgram/ml to values within the normal range (50-150 microgram/ml) were observed during 89% (25/28) of the favorable clinical responses which have been followed with C3DP assays. Increases in C3DP levels from values within the normal range to values above 150 microgram/ml were observed either prior to or coordinate with clinical symptoms of disease recurrence 83% of the time (10/12 cases). These studies suggest that serial C3DP determinations offer an excellent prognostic aid for evaluating the response of malignant tumors during chemotherapeutic management.


Combination hyperthermia and radiation therapy for cutaneous malignant melanoma. The clinical studies were carried out to evaluate whether radiation combined with hyperthermia would evoke better local tumor control than that achieved with radiation alone. Using an inductive method of radiofrequency heating at 27.12 MHz and radiation therapy, 37 lesions in 14 patients were treated with heat alone, radiation alone and heat in combination with radiation. In patients who completed the planned course of the combined therapy with an adequate follow-up period of more than six months, 16 out of 18 lesions (in 9 patients) had a complete response, whereas only 1 out of 8 lesions (in 5 patients) had a permanent regression after radiation alone. The amount of radiation dose necessary to achieve a satisfactory tumor control in combination with radiofrequency heating has been 3000 rad (300 rad X 10 in 4 weeks) to 3200 rad (400 rad X 8 in 4 weeks). Importance of the temperature measurements of both tumor and the adjacent normal tissue is discussed.


Delayed hypersensitivity and lymphocytic transformation in patients with Hodgkin's disease and granulomas. Noncaseating, sarcoid-like granulomas were found in the tissues of 9 out of 31 patients with Hodgkin's disease. In vivo and in vitro cell-mediated immunity was evaluated in patients with and without granulomas and compared to a group of 20 normal controls. Hodgkin's disease patients of both groups showed a significantly reduced in vivo and in vitro response when compared to the control group. However, when patients in stages IIIB and IV were eliminated and patients in stages I, II, and IIIA examined separately, a positive skin test response to one or more antigens was elicited in 85.7% of patients with granulomas, while a markedly decreased-dose dependent response was observed in patients without granulomas. In vitro lymphocyte blastic transformation by phytohemagglutinin (PHA) was severely impaired in both groups of patients as determined by dose-responses curves. These results indicate that Hodgkin's disease patients with granulomas have a significantly better skin test response than those without granulomas.


Electron and immunoelectron microscopic study of thyroidal medullary carcinoma. A correlated light, immunofluorescent, electron and immunoelectron microscopic study on the histogenesis and immunochemical nature of the tumor amyloid in medullary carcinoma of thyroid (MCT) was performed. The light microscopic finding of cytoplasmic inclusions in tumor cells was related ultrastructurally to a unique form of cellular degeneration composed of particulate and fibrillar matters, which could be the precursor of amyloid deposits. The immunofluorescent staining revealed calcitonin in tumor cells and in portions of amyloid. Ultrastructurally, the storage granules were morphologically heterogeneous and contained variable amount of immunoreactive calcitonin. Immunoelectron microscopy demonstrated disintegration of the granules with apparent discharge of calcitonin into the matrix of the above-mentioned particulate and fibrillary mass. Immunoreactive calcitonin was not shown on the mature amyloid fibrils nor in most of the granular matters closely associated with the amyloid. The results indicate that the amyloid in MCT, though related to the secretory products of the tumor cells, may not represent the hormonally active calcitonin molecule.


A qualitative and quantitative study of monocytes in patients with malignant solid tumors. Monocytes from 21 patients with cancer of the lung and cancer of the prostate were studied prior to treatment. The absolute circulating monocyte count, serum lysozyme levels and monocyte IgG surface receptors were normal at all stages of the disease. Monocyte chemotaxis was defective in 45% of the patients. Serum chemotatic factor inactivator(s) that inhibit chemotaxis of normal monocytes were detected in 90% of the patients. In two of four patients the chemotactic factor(s) disappeared following surgical removal of localized tumors. The results of the chemotaxis studies may explain the data of defective delayed hypersensitivity reactions frequently seen in patients with malignancies. The defective chemotaxis and the presence of chemotatic factor inactivator(s) may interfere with the ability of monocytes to accumulate as macrophages in tumor sites.


Hemidesmosome-like intercellular specializations in human meningiomas. Desmosomes, gap junctions and tight junctions are intercellular specializations which have been described in human meningiomas. We report, in five of five meningiomas, including a pulmonary metastasis from a malignant intracranial meningioma, a fourth type of intercellular specialization which is similar to but not identical with the hemidesmosome of the epidermis. Idential intercellular specializations have been described between arachnoidal cells, but not between dural fibroblasts. The presence of the junction in human meningeal neoplasms of diverse histologic pattern supports the origin of the meningioma from arachnoidal epithelium. The consistent occurrence of these structures in a pulmonary metastasis as well as benign intracranial tumors suggests that they may be a useful morphologic marker in the differential diagnosis of putative extracranial metastatic meningiomas.


Anaplastic carcinoma of the thyroid: a review of 84 cases of spindle and giant cell carcinoma of the thyroid. Eighty-four cases of spindle and giant cell carcinoma of the thyroid were reviewed. This is a swiftly growing, and rapidly fatal neoplasm that generally develops in the elderly. Treatment of this disease was generally unsuccessful with a 7.1% five-year survival rate and a mean survival period of 6.2 months from the time of tissue diagnosis. In the majority of cases, areas of well-differentiated thyroid carcinoma could be identified, supporting the concept that spindle and giant cell carcinoma results from the transformation of preexisting well-differentiated thyroid carcinoma. Because of the aggressive biologic activity of this neoplasm, we treat all cases as disseminated disease at the time of presentation. Our best therapeutic results have been obtained with a combination of surgery, irradiation, and chemotherapy; however, these results still leave much to be desired. Patients with only small foci of spindle and giant cell carcinoma, at the time of diagnosis, may have a better chance of prolonged survival.


Mechanisms of hypercalcemia in malignancy. Various hormones have been implicated in the genesis of hypercalcemia in patients with malignancy. Ectopic secretion of PTH by tumor has been documented in only a few patients; rather, elevated levels of circulating iPTH have been presumed to reflect tumor production of hormone in most patients. Small fragments of PTH, as well as polypeptides larger than native PTH, have been described; their biological roles are unclear. The pattern of immunoreactivity, however, has been used to differentiate patients with ectopic hyperparathyroidism from patients with concomitant primary hyperparathyroidism. Vitamin D-like sterols produced by breast cancer seldom reach plasma levels necessary for physiological effects. Members of the prostaglandin family have been proposed to induce hypercalcemia through osteoclast activation or alteration of the immune system and also to affect the frequency of bone metastases. At present, no direct evidence is available to prove a direct role for these effects and prostaglandins are most useful as possible indicators of disease activity.


Subretinal fluid examination of LDH, PGI, and CEA in a case of metastatic bronchogenic carcinoma of the choroid. A 58-year-old white male with a history of bronchogenic carcinoma presented with a total retinal detachment overlying a choroidal metastasis. His main tumor burden had been extirpated by pneumonectomy followed by radiation therapy four months prior to admission. With the development of a painful, glaucomatous eye, unresponsive to conventional therapy, enucleation was performed. Histologic examination of the enucleated globe revealed a metastatic tumor to the choroid, consistent with primary bronchogenic carcinoma. Aqueous humor and plasma examination revealed elevated ratios (Aqueous humor:Plasma) of lactate dehydrogenase (LDH) and phosphoglucose isomerase (PGI). Furthermore, subretinal fluid examination demonstrated concentrations of LDH and PGI higher than aqueous humor. While the level of carcinoembryonic antigen (CEA) in the plasma was normal (less than 2.5 ng/ml) following pneumonectomy, it was 121 ng/ml in the subretinal fluid. This would suggest that a choroidal lesion alone, in the absence of a clinically detectable primary tumor, is insufficient to elevate the plasma CEA.


Ewing's sarcoma: an autopsy study.

Ewing's sarcoma: an autopsy study. An autopsy study of 26 cases of Ewing's sarcoma treated with radiation to the primary site plus adjuvant chemotherapy has shown metastatic tumor in 23 cases. Metastases were found typically in lungs, pleura, bones and regional lymph nodes. In three cases no tumor could be found at autopsy, and death was due to complications of treatment. Tumor was found in the irradiated primary site in 13 of the 20 cases in which the primary site was examined at autopsy. Histologically, the tumor at autopsy frequently had increased pleomorphism and increased numbers of bizarre giant cells; however, these changes did not affect the presence of glycogen in tumor cells, thus reaffirming the importance of intracytoplasmic glycogen in the diagnosis of Ewing's sarcoma.


Increased in vitro tetraploidy: tissue specific within the heritable colorectal cancer syndromes with polyposis coli. In vivo expression of human hereditary tumors are known to be tissue specific; in familial polyposis coli the genotype is expressed solely as colonic polyps that become malignant and in the Gardner syndrome as extracolonic connective tissue tumors and related neoplasms in addition to such colonic lesions. In vitro such tissue specificity was also seen in these 2 syndromes. Increased tetraploidy has been observed only in those cultures derived from tissues, which although appearing normal in the patient, were known to undergo malignant transformation in vivo based on clinical phenotypes and family histories: colonic mucosa in familial polyposis coli, skin and colonic mucosa in the Gardner syndrome. Cultures established from tissues known not to show neoplastic growth or from benign tumors (fibromas, sebaceous cysts and lipomas) did not show increased tetraploidy. Increased tetraploidy in cultures established from these 2 syndromes did not identify all cultured cells with either mutant genotype or those cells showing abnormal benign growths in vivo but rather only in those that are known to undergo malignant transformation in vivo in both syndromes. Such observations suggested that in these 2 syndromes there was a population of tetraploid cells, at least in culture, constantly present which may be relevant to the multi-step process of carcinogenesis.


Gastric cancer in a coal mining region.

Gastric cancer in a coal mining region. A significant excess incidence of gastric cancer was previously reported in the coal mining counties of Utah for 1965-1969. A repeat study for the period 1970-1975 showed the rates to be close to expectation and are in line with the slight excess mortality reported by Creagan et al. from United States coal mining counties. A cluster of 12 cases out of 43 reported for the 11-year period occurred in 1965. Cases during 1966-1975 appear to have occurred at random by year. The previous suggestion that the high incidence of gastric cancer in Carbon and Emery Counties could be related to frequent exposure to coal-carrying hydrocarbons appears to be unwarranted.


Cardiac candidiasis in cancer patients.

Cardiac candidiasis in cancer patients. Histologically proven infection of the heart by Candida organisms occurred in 17 (10%) of 168 cancer patients with candidiasis studied at necropsy. All 17 patients were among the 85 patients with disseminated Candida infection; none of the 83 patients with localized candidiasis had involvement of the heart. Abscesses in the myocardium were present by histologic examination in all 17 patients and in 8 were evident on gross inspection as well. The mural endocardium was also affected in 5 patients from direct extension of a myocardial abscess. The valves were uninvolved. Pericardial infection was detected in 2 patients. The 17 patients with cardiac Candida infection had a higher frequency of positive premortem blood and cerebrospinal fluid cultures and of presence of Candida in the heart's blood than did the 68 patients with disseminated candidiasis but without involvement of the heart. Symptoms or signs of cardiac dysfunction resulting from the cardiac Candida infection were not readily detected in any patient. The clinical and pathologic features of cardiac candidiasis in cancer patients are distinct from those observed in Candida endocarditis.


Non-Hodgkin's lymphoma in children: an analysis of 122 cases from Argentina. One hundred twenty two children with non-Hodgkin's lymphoma were studied from January 1966 to December 1975. The first group (1966-1972) did not receive an uniform treatment. The second group (1973-1975) entered in a G.A.T.L.A. protocol consisting of: vincristine-prednisone plus surgery and/or radiotherapy as induction treatment, craniocervical radiotherapy and intrathecal methotrexate as CNS preventive treatment and anti-leukemia (6-mercaptopurine, methotrexate and vincristine-prednisone pulses) or anti-lymphoma (COPP) treatment as maintenance, in a randomized trial. Comparison of survival of the two groups are as follows: series 1966-1972, 22% and 20% at 12 and 24 months of evolution, respectively, and series 1973-1975, 33% and 26% at 12 and 24 months, respectively. After 2 years of complete remission we have not seen any relapse. We conclude that 1) this disease is highly malignant and must be treated with more intensive chemotherapeutic treatment, and 2) there is no difference between antileukemia or anti-lymphoma maintenance treatment, as yet.


Contacts among patients with hematological malignancies. A survey was carried out in 2 country areas of New South Wales with the aim of defining contacts among patients with lymphoma, leukemia and myeloma, and of determining whether these were more numerous than among matched controls from the same localities. Cases were identified from records of the N.S.W. Central Cancer Registry and of local doctors and hospitals. 184 cases were found, corresponding to the expected number, and 145 patients, as well as the same number of controls, were interviewed. Of the 290 patients and controls surveyed 111 (38.3%) had had one or more contacts with other patients or controls (37.9% of patients and 38.6% of controls). There were 24 case-case pairs involving contacts among 33 individual patients, 23 control-control pairs involving 36 individuals, and 38 case-control pairs involving 66 individuals. A statistical analysis using a weighting system showed that numbers, closeness and duration of contacts among patients and patients did not differ significantly from those expected. These results thus provide no evidence in favor of the hypothesis that the diseases were transmitted from patient to patient in the survey areas.


Diagnostic tests and Hodgkin's disease: a standardized approach to their evaluation. We have analyzed a series of 124 young men with Hodgkin's Disease in an attempt to clarify the role of non-invasive tests in the staging evaluation. Data are presented in a way which permits comparison between series and application of the data to decision-making models. Analysis of the data shows our patients to have had primarily limited (stage I and II) disease. In our hands certain tests were quite accurate in predicting the absence of abdominal disease (lymphangiography, hepatic scintigraphy) as determined by laparotomy, but no study appeared to eliminate the need for laparotomy in this group. Data from several other large series are also presented in the same format. The data demonstrate a large variability between institutions in the predictive value of a positive lymphangiogram. Other studies, such as splenic scintigrams, appear to have more consistent predictive values. We encourage further use of this type of data presentation.


Cowden's disease: a cutaneous marker of breast cancer. Cowden's diseases features facial trichilemmomas (a benign tumor of follicular epithelium), acral keratoses on the limbs, and oral mucosal papillomas and fibromas; it may also involve thyroid, gastrointestinal tract, ovaries, uterus, and breasts. Among 32 known cases of Cowden's disease, 21 are women, in 10 of whom breast cancer has already developed (bilateral in 4). The 11 women in whom breast cancer has not yet developed have fibroadenomas, fibrocystic disease, virginal hypertrophy of the breast, and malformations of nipples and areolae. Their median age is only 36 years. Two have mothers with breast cancer and in one both mother and maternal grandmother had breast cancer. Dermatologic lesions, including pathognomonic multiple facial trichilemmomas, precede the development of malignancy and can identify women with ahigh risk of developing breast cancer.


Fine needle aspiration biopsy of malignant lesions in and around the pancreas. A simple method of percutaneous fine needle aspiration biopsy of malignant lesions in and around the pancreas following radiological localization is described. Biopsy was performed in 28 patients. Twenty-three were eventually considered to have malignant disease and biopsy was positive in 19 of these (83%), including 16 of 18 patients with carcinoma of the pancreas (89%). One patient developed an exacerbation of pancreatitis following the procedure. Surgery became unnecessary in 5 of the 18 patients with positive biopsies. From these results and from a review of the literature on percutaneous fine needle aspiration biopsy, we believe that this is a simple, relatively painless, reasonably safe and reliable method of obtaining a preoperative diagnosis and advocate the procedure as desirable in all patients in whom malignant disease is suspected and can be localized radiologically.


Production of bone resorbing activity in poorly differentiated monocytic malignancy. A patient with poorly differentiated monocytic malignancy had extensive lytic lesions of bone and hypercalcemia. The patient's bone margins showed little evidence of osteoclast activity. Serum parathormone concentrations were slightly increased relative to the degree of hypercalcemia, consistent with impaired renal function. Serum concentrations of the relevant prostaglandins were normal. Supernatant medium from cultured tumor cells caused prominent bone resorption in vitro. The studies on this patient provide evidence that malignant cells of the monocyte-macrophage line are capable of direct bone resorption.


Synchronous malignant glandular schwannomas in congenital neurofibromatosis. An eight-year-old child with congenital neurofibromatosis developed rapidly growing synchronous malignant schwannomas in the neck and mediastinum. These tumors, displaying a biphasic pattern of spindle cell sarcoma admixed with mucinous epithelium, represent the eighth and ninth known examples of glandular schwannoma. The clinical and pathological features of this case are detailed and the histogenesis of the tumors discussed.


Intralesional treatment of recurrent metastatic cutaneous malignant melanoma: a randomized prospective study of intralesional Bacillus Calmette-Guerin versus intralesional dinitrochlorobenzene. Eighteen patients with multiple recurrences of malignant melanoma without evident distant spread were randomly assigned to treatment with either intralesional Bacillus Calmette-Guerin (BCG) or intralesional dinitrochlorobenzene (DNCB). Both agents were able to destroy approximately 90% of the injected intradermal nodules. Intradermal disease was more easily obliterated than subcutaneous disease with intralesional treatment with either agent, and local control of satellitosis with elimination of all clinically evident tumor was achieved in the patients who had intradermal without subcutaneous satellitosis, regardless of whether the patient was receiving BCG or DNCB. The clinical courses of the treated patients were essentially the same. Although PHA reactivity was depressed, the patients in both groups were responsive to recall and melanoma skin test antigens, demonstrated leukocyte migration inhibition with melanoma antigen and were generally within normal limits when assayed for 29 degrees C E rosettes. Our study demonstrated a dramatic difference in toxicity between the two intralesional agents without a similar difference in therapeutic efficacy or immune testing.


Adenocarcinoma of the uterine cervix in Jewish women: a distinct epidemiological entity. During an 11-year period, 532 Jewish women with invasive cancer of the uterine cervix were diagnosed in Israel. Of these, 441 (82.9%) had squamous cell carcinoma (SUC) and 52 (9.8%) had adenocarcinoma (AUC). Different characteristics were noted for these two histological categories with regard to age, ethnic distribution, and chronological time trends. The ratio between AUC and SUC differed considerably between the various ethnic groups, ranging from about 1:6 in the Asian and European born to less than 1:14 in the North African. This indicates that the relatively higher incidence of AUC in Israeli Jewish women is limited to those ethnic groups that have a low risk for cervical cancer. The different patterns of SUC and AUC in the population support the contention of a varying etiology.


Familial intraocular malignant melanoma: a case report. A family with histopathologically proven intraocular malignant melanoma involving two generations is reported. A member of third generation was likely involved by history. Intraocular malignant melanoma, its familial occurrence, its occurrence with multiple malignancies, and its relationship to multiple malignancies occurring in a family are discussed.


Metastasis via the blood stream: the method of intravasation of tumor cells in a transplantable melanoma of the hamster. Tumor cells entered the vascular compartment of transplantable amelanotic melanoma of the hasmster 3 days after transplantation, closely following vascularization of the transplant. Examination of the tortuous giant capillaries at the edge of the transplant showed tumor cells breaching the thin endothelium of these vessels. Extravasation of red cells into the surrounding tumor tissue and attempted haemostasis by small platelet aggregates attached to the vessel walls were also found.


Establishment and characterization of two hamster macrophage cell lines. Two hamster macrophage cell lines (HM-1 and HM-2) were established in vitro from lymphoid tumors produced in hamsters by direct implantation of normal human umbilical cord leukocytes. Despite long-term culture, both cell lines maintained the morphological, functional and surface characteristics of normal macrophages. It is considered that these cell lines were derived from host macrophages infiltrating the heterotransplants.


Degradation and intracellular phosphorylation of showdomycin. The anti-tumor agent showdomycin is degraded in the presence of 0.05 M NH4HCO3 (pH 8.0) to a compound which carries a negatively charged group. Together with previous data (Darnall et al., 1967) it is suggested that ammonia causes a basic hydrolysis of the maleimide moiety under formation of a maleic acid-like compound. During this modification the C-glycosidic bond is not split. Intracellularly, in mouse lymphoma cells (L5178y), showdomycin is phosphorylated to showdomycintriphosphate, via the mono- and diphosphate stage.


Carcinogenicity of 8-nitroquinoline in Sprague-Dawley rats. 8-Nitroquinoline was added to the semisynthetic diet of outbred Sprague-Dawley rats for 48 weeks. Squamous cell carcinomas of the upper digestive tract (tongue and esophagus), but especially of the forestomach, occurred among the higher concentration groups. Squamous cell carcinoma of the forestomach occurred in 3 of 11 (27.3%) rats in the 2500 ppm group and 7 of 35 (20.0%) rats in the 1000 ppm group. Three cases of lung adenoma were also observed in the 2500 ppm group. Results clearly indicate that 8-nitroquinoline is carcinogenic in rats when administered orally in a semisynthetic diet.


Enhancement of mutagenesis during cell replication of cultured liver epithelial cells. The susceptibility to mutagenesis of proliferating and non-proliferating mammalian cells was studied in cultured rat liver epithelial cells. Cells brought to growth quiescence by a non-toxic means were stimulated to proliferate and both types of cultures were exposed to methyl methanesulfonate (MMS). Cultures enriched in proliferating cells were more susceptible to both the toxic and mutagenic action of the mutagen than were quiescent cultures with a low level of proliferation.


Size dependence of cell cycle parameters for lung metastases from B16 melanoma. Data presented show that the cell cycle time of the lung metastases from a B16 melanoma tumor growing in the foot pad of C57 BL/6J mice increases as the size of the metastases increases. This increased cell phase durations of the small metastases being closer to those of the primary tumor. The growth fractions of the primary tumor and of the lung metastases of different sizes are analyzed.


A new and advantageous model for colorectal cancer: its comparison with previous models for a common human disease. Weekly subcutaneous injection of 10 mg/kg body wt. of N-nitrosobis(2-oxopropyl)amine (BOP), a pancreatic carcinogen in Syrian golden hamsters, induced colorectal cancer in MRC rats in 67% of all males and 33% of all females that survived beyond 43 and 68 weeks, respectively. Tumors were concentrated in specific segments of the large bowel and not found in the small intestine. Although the distribution of cancer in the cecum, and ascending and descending colon was similar in both sexes, rectal cancer predominated significantly in males. These data and the tumor morphology indicate the present model more closely resembles the corresponding human disease than do models in previous relevant experimental studies.


Methylnitrosophenylurea--a highly carcinogenic compound. In vitro, 1-methyl-1-nitroso-3-phenylurea (MNPU) was easily formed from 1-methyl-3-phenylurea (MPU) and sodium nitrite in an acid environment. In rats MNPU showed a strong carcinogenic effect, inducing tumors of the forestomach. MNPU was formed endogenously after combined administration of MPU and nitrite to rats, as indicated by the induction of forestomach tumors. MPU itself was not carcinogenic. In the metabolism of the herbicide 1,1-dimethyl-3-phenylurea (fenuron) MPU resulted as a demethylation product. The possible significance for preventive oncology and the role of MNPU as an environmental carcinogen are briefly discussed.


Interactions between polonium-210 alpha-radiation, benzo(a)pyrene, and 0.9% NaCl solution instillations in the induction of experimental lung cancer. Benzo(a)pyrene and 210Po were administered both simultaneously and sequentially by intratracheal instillations to Syrian golden hamsters in experiments designed to identify any synergistic interactions between these carcinogens. Their effects were additive after simultaneous administration. A significant synergistic interaction between the two agents appeared to occur when benzo(a)pyrene exposure followed 4 months after 210Po exposure. Most of this effect could be ascribed, however, to a potentiating effect of subsequent 0.9% NaCl solution instillations on 210Po carcinogenesis. These results emphasize the fact that seemingly innocuous stimuli may significantly potentiate lung carcinogenesis. The implications of these findings in terms of the interactions between alpha-radiation and cigarette smoke in human populations are discussed.


Immobilized carboxypeptidase G1 in methotrexate removal. Carboxypeptidase G1, and enzyme capable of cleaving the glutamate moiety from a variety of folate analogs, has been immobilized on nylon tubes and hollow fibers for use in extracorporeal enzyme reactors for methotrexate (MTX) removal from blood. The stability and reactor parameters of the system have been investigated with the use of single tubes and a multitubular arrangement. The results are used to predict their MTX-removing capacity at flow rates and MTX concentrations of clinical interest. In vivo experiments with dogs demonstrate the potential applications of such extracorporeal shunts in "rescue" after administration of large doses of MTX. For dogs a carboxypeptidase G1 reactor with a clearance value of about 150 ml/min would be required in such application.


Uncharged nuclear receptors for estrogen in breast cancers. This study was undertaken to define the incidence and concentration of uncharged nuclear estrogen receptors (RN) in human breast cancer. The concentrations of RN and cytoplasmic uncharged receptor were determined on sucrose gradients following a 4-hr incubation at 4 degrees with 1.6 nM 17 beta-[3H]estradiol in 139 tumor specimens from 137 patients. RN was extracted from washed nuclear pellets in buffer containing 0.4 M KCl. The receptor molecule extracted had a high affinity for 17 beta-[3H]estradiol (Kd = 0.9 to 7.6 nM) and was specific for estrogen. The possibility of artifact due cytoplasmic contamination of the nuclear fraction or high-ionic-strength-induced exchange of charged nuclear receptors was rendered unlikely by validation experiments performed with pooled tumor tissue. Significant amounts of cytoplasmic uncharged receptor (greater than 7 fmol/mg protein) were found in 63.3% of the tumors. Similar significant amounts of RN were found in 29.5% of the tumors. Significant amounts of RN in the presence of undetectable cytoplasmic uncharged receptor were found in 2.9% of the tumors. The percentage of tumors that contain significant amounts of RN is approximately the same percentage of estrogen receptor-positive tumors that do not respond to ablative therapy.


Light and electron microscope study of osmotically induced tumor necrosis. Necrosis of 1.5-cm Walker 256 tumors was produced by injecting a strongly hypertonic solution of glucose in and around the growths and by delaying resorption of the solution with serotonin, given s.c. at a distance. The morphological changes occurring in 13 tumors were followed by light and electron microscopic analysis of samples taken 0.25, 0.5, 1, 1.5, 3, and 5 hr after treatment. The 0.25-hr samples showed mitochondrial swelling, loss of cristae, and flocculent material within the inner compartment. Swelling of the mitochondria persisted in the 0.5-hr specimens (as it did in all subsequent samples), and it was accompanied by clumping and margination of chromatin. These changes were more pronounced at 1 hr, at which time the nuclear and plasma membranes were frequently ruptured. The endoplasmic reticulum and Golgi complex could no longer be recognized. The 3-hr material revealed ruptures of the outer mitochondrial membrane with myelin figures and discontinuous cell membranes. In the 5-hr samples, the nuclei exhibited a dark nucleoplasm and large clumps of chromatin. The perinuclear membrane was not always recognizable.


Histological conformity of implantation tumors produced by kidney cell lines derived from dimethylnitrosamine-treated rats, with dimethylnitrosamine-induced renal mesenchymal tumors. The histology of five implantation tumors induced in rats by the deposition of cultured cell lines derived from dimethylnitrosamine (DMN)-treated rats is described and compared with the morphology of the predominant kidney neoplasm induced in vivo by a single high dose of DMN. The cell lines leading to growth upon implantation were long-established, continuously growing cultures obtained either from a DMN-induced renal mesenchymal tumor or from rats treated shortly before with a carcinogenic dose of DMN. The latter cultures had expressed morphological transformation at subcultures 5 or 6. All of the implantation tumors were of mesenchymal type, comprising variously a range of cell forms including fibroblast-like spindle cells, smooth muscle fibers, and "giant" cells, which resembled common aspects of the parent mesenchymal tumors induced in the rat kidney by DMN. Deposition of cells intrarenally illustrated the survival of remnants of preexisting nephrons as epithelial profiles scattered through the proliferating malignant tissue, a feature most characteristic of the parent tumor. The results confirmed the malignant nature of the various cell lines tested, in keeping with their altered behavior in vitro, and they were consistent also with the premise that the in vivo-in vitro system is selecting cells in culture that represent the same target population from which the renal mesenchymal tumors are derived in vivo.


Isolation of a bone-resorptive factor from human cancer ascites fluid. A protein fraction that induces the resorption of bone explants in organ culture was isolated from the ascitic fluid of patients with advanced cancer metastatic to the peritoneal cavity. Partial purification was achieved by means of gel filtration, affinity chromatography, and ion-exchange chromatography. The isolated fraction, the components of which have an apparent molecular weight of 60,000, was found to be heterogeneous by disc gel electrophoresis and to be composed primarily of proteins with relatively acidic electrophoretic properties. The specific bone-resorptive activity of this protein fraction was greatly increased over that of the unfractionated starting material, and the activity could be completely destroyed upon incubation with pronase and on heating. As determined by immunoassay and extraction procedures with various solvents, the bone-resorptive action of the isolated fraction was not attributable to the presence of parathyroid hormone, prostaglandin E2 or vitamin D-like sterols. In parallel experiments the supernatants of phytohemagglutinin-stimulated normal human peripheral leukocytes were subjected to identical chromatographic techniques, and a proten fraction with a molecular weight of 60,000, which resembled the resorptive fraction isolated from cancer ascites fluid and which contained significant bone-resorptive activity, was also partially purified.


Soluble suppressor factor from the spleens of tumor-bearing mice. Spleen cells from tumor-bearing mice when cultured for 3 to 5 days released a soluble factor into the media that suppressed the stimulation of lymph node and spleen cells by tumor antigen or mitogens. Spleens from mice bearing MC43 tumors for 14 days were capable of producing suppressor factor in vitro, while those from mice bearing the tumor for 10 days or less failed to do so. Lymph node cells from the same animals did not produce suppressor factor in vitro. The suppressor factor was produced by a nonadherent cell population, was heat stable, was lost on dialysis, and did not appear to be tumor antigen or thymidine.


Increased activity of rat liver messenger RNA and of albumin messenger RNA modulated by thioacetamide. Administration of thioacetamide to rats was found to increase the activity of liver messenger RNA in the wheat germ protein synthesis assay. The synthesis of total protein was increased about 2.5-fold after 4 days of treatment. The treatment was associated with an increase in the relative quantity of polyadenylic acid-containing RNA. Immunoprecipitation studies showed that albumin synthesis directed by the messenger RNA was increased disproportionately, reaching a level 5 to 6 times the control after 4 days of thioacetamide administration.


Effect of cholecystoduodenostomy and choledochostomy in pancreatic carcinogenesis. The pancreatic carcinogen N-nitrosobis(2-oxopropyl)-amine was administered to Syrian golden hamsters after cholecystoduodenostomy and choledochostomy to investigate a possible bile reflux mechanism as a factor in pancreatic carcinogenesis. The induced lesions were similar in morphology, multiplicity, and distribution to those of animals in other studies without surgery. Hence the findings contraindicate the importance of biliary reflux in pancreatic tumor induction. Other possible mechanisms are discussed.


Enhanced response to chemoimmunotherapy and immunoprophylaxis with the use of tumor-associated antigens with a lipophilic agent. Treating iodoacetamide (IAD)-modified lymphoma cells with the lipophilic agent dimethyldioctadecylammonium bromide (DDA) increased their immunogenicity as evidenced by the increased capacity of syngeneic, vaccinated hosts to reject subsequent implants of the same lymphoma. Under conditions of suboptimal immunization to facilitate comparison, there were 61% survivors among mice challenged with tumor implants after immunization with modified cells and DDA compared to 20% survivors among those immunized in the absence of DDA. The enhanced immune response was dependent on DDA dosage and was most striking when DDA was directly complexed to the IAD-treated cells. DDA was also effective with solubilized tumor antigen and with lymphoma cells not pretreated with IAD, but the latter had to be heat killed to assure that they were nontumorigenic. In therapy experiments BALB/c mice bearing P1798 were treated with methotrexate followed by immunotherapy with IAD-P1798 alone or complexed to DDA. With two and three cycles of therapy, methotrexate alone yielded 5 and 13% survivors, while adding immunotherapy with the DDA complex gave survival rates of 63 and 71%. In the absence of DDA, chemoimmunotherapy with methotrexate and IAD-P1798 gave intermediate results. In the absence of antigen, DDA was ineffective in either immunoprophylaxis or therapy experiments.


Effects of hydroxyurea on murine type C virus-specific DNA synthesis in newly infected cells. Cell transformation and replication of the Rauscher pseudotype of Moloney murine sarcoma virus in mouse embryo fibroblasts were inhibited by hydroxyurea within a critical time period of 30 to 90 min postinfection. In cells infected by Rauscher leukemia virus, treatment with 1mM hydroxyurea during the critical time period resulted in the accumulation of minus-strand DNA (molecular weight, 3 x 10(6)) in association with the parental viral genoma RNA. This 5 to 6 x 10(6) dalton RNA:DNA hybrid was found in the cytoplasm. Positive-strand DNA of genomic or smaller size was not detected in the presence of hydroxyurea, but virus-specific DNA was found in the nucleus 30 min after removal of drug.


Suppression by Nocardia rubra cell wall skeleton mammary DNA synthesis, plasma prolactin level, and spontaneous mammary tumorigenesis in mice. The effects of the cell wall skeleton of Nocardia rubra on mammary gland DNA synthesis, plasma prolactin levels, and spontaneous mammary tumorigenesis in mice were studied. Female SHN mice received s.c. injections of 100 microgram N. rubra cell wall every 7 days between 2 and 12 months of age. The treatment resulted in the marked inhibition of mammary tumorigenesis; incidence was significantly lower in the experimental mice than in the controls except at 9 and 12 months of age. The age of onset of mammary tumors was significantly higher in the former than in the latter. In association with these findings, the treatment also reduced normal mammary gland DNA synthesis and prolactin levels in the circulation, both of which are primary factors for mammary tumorigenesis.


Studies of the growth, population kinetics, and host lethality of CD8F mammary adenocarcinoma. First-generation transplants of spontaneous mammary adenocarcinomas in CDSF mice have been included in the tumor panel used for the screening of potential anticancer chemotherapeutic agents. Studies were undertaken to evaluate the growth and kinetic characteristics to provide a better understanding of the basic biology of the tumor system. The spontaneous mammary adenocarcinomas are highly variable in growth rate, and this characteristic is seen to a lesser extent in the transplanted tumors. The doubling time of the transplants is usually, but not always shorter than the doubling time of the donor tumor at a similar mass. The median length of the cell cycle obtained from pooled data from first-generation transplants of four donor tumors is 15.6 hours with a median S phase of 9.5 hours. The thymidine indices (TI) ranged from a mean of 0.28 at 100 mg to a mean of 0.20 when tumors ranged from 3 to 6 g. The mean TI of tumors of all sizes is 0.27. The cell cycle parameters and TIs appear to be characteristic of the tumor system and apparently do not vary with characteristics of the donor tumor. These studies also indicate that these kinetic parameters are not responsible for the variation in volume doubling time of the transplanted tumors.


Analysis of methotrexate and 7-hydroxymethotrexate by high-pressure liquid chromatography. The use of high-dose methotrexate (MTX) therapy in cancer chemotherapy has intensified the need to monitor the purity of this drug. A high-pressure liquid chromatographic method, which utilizes ion-pairing on a mu-Bondapak C18 column, has been developed for this purpose. This method provides an efficient and highly sensitive method for detecting impurities in commercially available MTX. As has been demonstrated previously, patients receiving high-dose MTX therapy metabolize some of this drug to 7-hydroxymethotrexate (7-hydroxy-MTX), a derivative which is considerably less soluble than MTX itself. Consequently, 7-hydroxy-MTX may contribute to the renal toxicity commonly associated with high-dose therapy. An assay similar to the one described above is also applicable to this problem and can be used to assay MTX and 7-hydroxy-MTX directly in patients' urine.


Qualitative and quantitative toxicity of single and sequential sublethal doses of 5-fluorouracil in BDF mice. Qualitative and quantitative toxicity studies were conducted in BDF mice after ip injection of 33, 66, 134, and 200 mg/kg of 5-fluorouracil (5-FU) in daily doses for 1, 2, 3, or 4 consecutive days. Groups of five mice from each schedule and dose level provided blood and selected visceral organs for hematologic and histologic evaluation on posttreatment Days 1, 3, 6, 10, 14, and 21. Comparable groups of five mice were blood donors for biochemical evaluation of pooled plasma samples. The results indicated the onset, severity, and duration of injury to target organs and characterized the relationship of sublethal toxicity to drug dose and frequency of administration. Marrow suppression was reflected in peripheral erythropenia, thrombocytopenia, and leukopenia and also in medullary erythropenia and granulocytopenia. A transient elevation of plasma LDH, loss of body weight, and appearance of microscopic lesions of the gastrointestinal mucosae were additional indices of toxicity. The BDF mouse appears to provide a good qualitative and quantitative prediction of 5-FU toxicity, although the mouse apparently is quantitatively more sensitive than man.