Chlorpropamide-alcohol flushing: a definition of its relation to non-insulin-dependent diabetes. The single-challenge test for chlorpropamide-alcohol flushing (CPAF) was used to study two groups of patients with non-insulin-dependent diabetes and a family history of the disease who were distinguished only by their age at diagnosis (under and over 30). Their relatives were also studied. The proportions of patients showing CPAF in both groups were similar, and the family histories suggested dominant inheritance. When offspring of diabetics in whom the disease was diagnosed early were studied CPAF seemed to precede the appearance of diabetes. We conclude that the patients in both groups had the same, distinct syndrome, which is characterised by diabetes diagnosed at any age that is inherited as an autosomal dominant trait and associated with CPAF. This syndrome, which constitutes about one-fifth of all cases of non-insulin-dependent diabetes, may be detected with a single-challenge CPAF test before the onset of glucose intolerance. CPAF therefore acts as a genetic marker for the syndrome.


Myocardial ischaemia in patients with frequent angina pectoris. One hundred patients with angina pectoris underwent 16-point electrocardiographic (ECG) mapping of the left hemithorax during a standardised exercise test. Forty-five patients had maximum ST-segment depression at position V5, while 35 had no ECG signs of ischaemia at this position. In 20 V5 was on the edge of the precordial area, which showed less severe ST-depression than the central positions. An Oxford ECG recorder and highspeed analyser were modified and used in 50 of the patients with daily angina for recording ST-segment changes over 24 hours. Serial 24-hour ambulatory recordings from the edge of the precordial area of ischaemia identified during exercise detected a mean of only 14 +/- SD 3% of the episodes of ST-segment changes recorded from the centre of the same area. Only 16 +/- 2% of the episodes detected by ECG were accompanied by chest pain. More episodes occurred between 4 am and 6 am than at any other time during the night. This study shows the importance of recording ECG evidence of ischaemia from the precordial position showing maximum changes during exercise. ECG evidence of ischaemia occurs more frequently than anginal pain. These objective measurements add important information to the frequency of chest pain reported by patients with ischaemic heart disease.


Precordial exercise mapping: improved diagnosis of coronary artery disease. Surface mapping of the exercise electrocardiogram (ECG) provides a measure of the precordial area, severity, and time course of ST-segment changes occurring after exercise. Sixteen-lead isopotential surface maps were recorded before and after exercise in 109 patients with probable angina who subsequently underwent coronary arteriography. In addition, exercise ECGs with three orthogonal leads were obtained in 53 of these patients, and with a single unipolar chest lead in all 109. Of the 109 patients, 85 had significant (greater than or equal to 70%) narrowing of at least one major coronary artery. The sensitivities of the precordial surface mapping, orthogonal leads, and single chest lead (V5) when compared with the findings at coronary arteriography were 95%, 68%, and 64% respectively, while the specificities of the three lead systems did not differ significantly. The technique of precordial surface mapping after exercise may easily be applied in clinical practice and requires only conventional equipment available in most hospitals. It effectively aids diagnosis of coronary artery disease.


Effects of propranolol and metoprolol on the peripheral circulation. The effects of single doses of propranolol and metoprolol on skin temperature and skin and muscle blood flow were compared in 10 normal subjects and four patients with essential hypertension. In normal subjects the mean skin temperature fell by 1.30 +/- 0.62 degrees C 90 minutes after 80 mg propranolol and 0.15 +/- 0.05 degrees C after 100 mg metoprolol. Skin blood flow and resting muscle blood flow were not affected by metoprolol but fell significantly after propranolol. Both drugs reduced post-exercise muscle hyperaemia, propranolol by more than metoprolol. Similar changes were seen in the hypertensive patients. Propranolol should be used with care in patients with known vascular disease.


Liver enzyme concentrations as measure of possible infectivity in chronic asymptomatic carriers of hepatitis B. Fifty-two British-born blood donors who were chronic carriers of hepatitis B surface antigen (HBsAg) were tested for the presence of hepatitis B e antigen (HBeAg) and antibody to HBeAg by an immunoradiometric assay. The presence of HBeAg was closely associated with a slight rise in serum liver enzyme concentrations, a high HBsAg titre, and male sex. We suggest that the finding of persistently raised serum liver enzyme concentrations in an asymptomatic HBsAg carrier might be useful as a likely indicator of HBeAg and high infectivity.


Haemodynamic effects of buprenorphine after heart surgery. The effect of buprenorphine on the cardiovascular system was examined in 11 patients during the period of reduced cardiac reserve after open-heart surgery. Within 10 minutes of giving the full analgesic dose (5 microgram/kg) intravenously the mean heart rate had fallen significantly by six beats/min. Although in two patients the mean arterial pressure fell by 24 mm Hg, there was no overall change in mean arterial pressure, cardiac output, or peripheral resistance. In a further six patients buprenorphine was used successfully as the sole analgesic after open-heart surgery. Buprenorphine appears to be safer than morphine for use in patients with reduced cardiac reserve and is of similar analgesic efficacy.


A miniature Wright peak-flow meter.

A miniature Wright peak-flow meter. A new miniature Wright peak-flow meter has been designed and produced. The meter is tubular with a spring-loaded piston and a longitudinal slot through which air escapes. Its dynamic characteristics have been carefully designed to make it respond only to peak flow and not to rate of rise. Performance tests on early instruments showed fairly close correlation with the Wright peak-flow meter but with a constant error of + 38 1/min. On later models the correlation was increased to 0.990 and the error reduced to + 3%. The mini-meter correlates as well with the standard instrument as two standard instruments correlate with each other and should prove useful clinically.


Neurogenesis in spinal cord of mouse: an autoradiographic analysis. An autoradiographic analysis of the time and sites of origin, and the migration and settling patterns of neurons was made in the spinal cord of the mouse. The neurons originated on days 10--14 of gestation with temporal gradients along the ventrodorsal and rostrocaudal axes. The motor neurons originated on days 10 and 11 of gestation; the neurons in the intermediate gray region originated on days 11--14 of gestation; the neurons of the head of the dorsal horn originated on days 12--14 of gestation. The neurons that originated on days 10 and 11 originated and migrated primarily from the basal plate, and they settled in the adjacent regions of the intermediate zone; those neurons formed on days 12--14 originated and migrated primarily from the alar plate, and it was concluded that these neuroblasts similarly settled in the adjacent regions of the intermediate zone. Extraventricular proliferation, which presumably signaled the initial stages of gliogenesis, was first observed on day 12 of gestation. This study supports the classical idea of the mosaic pattern of neurogenesis in the embryonic spinal cord.


Messages conveyed by descending tracts during scratching in the cat. II. Activity of rubrospinal neurons. (1) The activity of rubrospinal (RS) neurons giving axons to the lumbosacral spinal cord was recorded during actual and fictitious8 scratching in thalamic cats. (2) During both actual and fictitious scratching, the discharge frequency of many RS neurons was rhythmically modulated. Different neurons were active in different parts of the scratch cycle, but most neurons were active in the flexor phase. (3) The discharge frequency within the bursts during fictitious scratching was, on the average, equal to that during actual scratching. Immobilization usually resulted only in a small displacement of the burst position in the scratch cycle. Therefore, rhythmical modulation of RS neurons is determined mainly by central mechanisms and not by a rhythmical sensory input. (4) In decerebellate cats, the overwhelming majority of RS neurons had no rhythmical modulation. Very weak modulation was found only in a few neurons. (5) Transection of the ventral spinocerebellar tract (VSCT) resulted in considerable reduction or complete cessation of rhythmical modulation in RS neurons during fictitious scratching. On the contrary, transection of the spino-reticulocerebellar pathway (SRCP) resulted in just a small decrease of modulation. Therefore, of the two pathways (the VSCT and SRCP) transmitting messages about intraspinal processes to the cerebellum during scratching2,3, the VSCT is of major importance for modulating RS neurons.


Interrelated changes of amino acids in the retina and optic tectum of a marine fish with alterations of illuminating conditions. Examination of interrelated changes in the concentrations of 'clusters' of amino acids provides evidence that individual amino acid levels in the tissues cannot be considered to vary independently. Such variations are observed when the CNS at 'rest' is stimulated. Alterations in taurine cause changes in glutamic acid and also modify its anatomically related compartmentalized metabolism. Similar interdependent modifications may be indicated for the concentrations of 3 biochemically related neutral amino acids: threonine, serine and glycine. With specific reference to the fish optic tectum and retina, certain differences and coincidences in the regulation of GABA and glycine have also been uncovered. Finally, many of the findings presented here are closely analogous to results obtained when comparing normal and epileptic mammalian tissues. Possibly, this analogy can be attributed to the fact that the physiological states so far investigated, from the most simplistic point of view, represent extreme variations of nervous tissue excitation.


The reciprocal synapse in the external plexiform layer of the mammalian olfactory bulb. It has recently been claimed by Ramon-Moliner29, that reciprocal synapses between mitral (and tufted) cell dendrites and granule cell gemmules, which for more than 10 years have been thought to represent a major feature of the structural and functional organization of the olfactory bulb28, are non-existent or, at best, extremely rare. The challenge is based on the contention that there is little, if any, morphological evidence to substantiate the existence of the gemmulofugal component of the reciprocal synapse. We have re-examined by electron microscopy the external plexiform layer (EPL) of the olfactory bulb of adult rabbits and rats. The study was carried out in conventionally prepared material, using individual and serial sections, goniometry, and quantitative analysis of montages, and also in material stained by the E-PTA and BIUL methods. According to currently accepted morphological criteria, gemmulofugal synapses can be identified consistently and with confidence at all levels in the EPL: they are particularly clearly and unambiguously resolved in E-PTA and BIUL preparations. At least the majority of gemmulofugal and mitrofugal synapses are closely associated in reciprocal pairs, as many previous authors have reported. Furthermore, there are approximately as many gemmulofugal as mitrofugal synapses in the EPL, which supports the long-standing proposal that the overall synaptic relationship between granule cells and mitral cells is a reciprocal one. We conclude that the  challenge of Ramon-Moliner to the reality and significance of the reciprocal synapse is not well-founded.


High-affinity GABA and glutamate transport in developing nerve ending particles. The high-affinity transport of [3H]GABA and [14C]glutamate was measured in discrete nerve ending fractions isolated from the developing rat cortex, beginning on day 7 postpartum and prepared using discontinuous Ficoll-isotonic sucrose gradients. On day 7 the material sedimenting in the lightest gradient fractions contained the highest density of particles capable of high-affinity transport. With increasing age, the transport sites became progressively associated with more dense particles in the gradient. On day 14, the Vmax values for both [3H]GABA and [14C-A1glutamate transport in the most dense nerve ending fraction were significantly increased over the adult value. This transient increase in the Vmax values disappeared by day 21. While the Km values for both transport systems were constant in all fractions for the adult animal, this was not the case in the developing pup. The Km values in the lightest fractions were significantly greater than those in the more dense fractions, suggesting that different transport systems may be present during development. Electron micrographs of the various fractions were prepared from 7-day and adult animals. These data illustrate that the lightest fractions of the '7-day' gradient are enriched in profiles similar to developing nerve endings. With development, these light fractions become infiltrated with myelin, the immature nerve ending profiles disappear, and the bulk of the nerve endings are found in more dense fractions.


The plasma membrane of bulk-isolated mature spinal neurons. Large neuronal perikarya isolated in bulk fractions from adult bovine ventral spinal cord and examined by electron microscopy have discontinuities in their plasma membrane. A systematic investigation of the possible causes of the membrane defects indicates that the act of mechanical dissociation of the neurons from the spinal tissue is chiefly responsible for the changes. One likely cause of membrane loss is the avulsion of postsynaptic membrane along with the presynaptic elements, leaving bouton-free cell bodies with many gaps in their plasma membrane. Membrane defects were also demonstrated by the entry of procion yellow into the isolated perikarya. Other studies29 on these neuronal cell bodies have also shown that many soluble proteins can leave the neurons under some, but not all, conditions of isolation. These findings are discussed in relation to the possibilities of tissue culture of mature neurons and future research on bulk-isolated mature neurons.


In vivo voltammetry: monitoring of dopamine metabolites in CSF following release by electrical stimulation. An in vivo electrochemical system which continuously records the concentration of metabolites of biogenic amines in small animal CSF is described. A small electrode, immersed in lateral ventricle CSF through a guide cannula, measures the amine metabolites by voltammetric oxidation. The detailed results of HVA release following electrical stimulation of the nigrostriatal pathway in rats are presented and compared with previous perfusion data. All the electrochemical results are verified by independent liquid chromatographic (chemical) analysis.


Receptive field development in the dorsal lateral geniculate nucleus in rabbits subjected to monocular eyelid suture. Receptive field properties of cells in the dorsal lateral geniculate nucleus (LGNd) were examined in 3 groups of rabbits, each subjected to monocular visual deprivation by lid suture at differing age periods. Monocular deprivation occurring from 6--8 to 20--25 days of age affected the normal development of LGNd receptive fields. A significant proportion of the cells in the deprived LGNd were either unresponsive to visual stimulation or had vague, indefinite receptive fields. Significantly fewer cells with uniform fields were found in the deprived LGNd than in the control. Percentages of concentric, motion and directional cells did not differ between the deprived and control LGNd. The diameters of receptive fields for concentric cells with sustained response properties, however, were smaller in the deprived than in the control LGNd. When deprivation was continued to 87--121 days of age, the percentage of uniform, indefinite and non-responsive cells found in the deprived LGNd approached more normal levels. Percentages of concentric, motion and directional cells were also normal. Monocular deprivation commencing at 21--22 days of age also had disruptive effects on LGNd receptive field organization, as reflected in the lower percentage of uniform and increased percentages of indefinite and non-responsive cells. These deficits, however, were not as severe as those seen in the animals deprived at an early age. A fourth group of adult rabbits subjected to monocular lid suture showed no such detrimental deficits in receptive field organization. These results demonstrated that visual deprivation affects the predominantly monocular LGNd of the rabbit, and that a critical period exists for such effects.


Messages conveyed by descending tracts during scratching in the cat. I. Activity of vestibulospinal neurons. (1) The activity of vestibulospinal (VS) neurons giving axons to the lumbosacral spinal cord was recorded during scratching in thalamic and decerebrate cats. The most part of the experiments was carried out on curarized cats, in which fictitious scratching13, i.e. rhythmical activity of motoneurons typical of actual scratching, was evoked. (2) During both actual and fictitious scratching, the discharge frequency of many VS neurons was rhythmically modulated in relation to the scratch cycle. Most modulated neurons were maximally active in the extensor phase of the cycle. (3) The firing pattern of VS neurons during fictitious scratching was similar to that during actual scratching. Therefore, rhythmical modulation of VS neurons is determined mainly by central mechanisms and not be a rhythmical sensory input. (4) In decerebellate cats, rhythmical modulation was not found during either actual or fictitious scratching. (5) Transection of the ventral spinocerebellar tract (VSCT) resulted in considerable reduction of rhythmical modulation of VS neurons during fictitious scratching, while transection of the spino-reticulocerebellar pathway (SRCP) resulted in just a small decrease of modulation. Therefore, of the two pathways (VSCT and SRCP) transmitting messages about intraspinal processes to the cerebellum during scratching6,7, the VSCT is of major importance for modulating VS neurons.


A chemical study on the development of the human forebrain and cerebellum during the brain 'growth spurt' period. I. Gangliosides and plasmalogens. Following upon previous studies on the lipid composition of the developing human brain, a further study is presented with the main object of tracing the chemical changes underlying the period of brain 'growth spurt'. Gangliosides and plasmalogens were selected as approximate markers of synaptogenesis and myelination, respectively, and these lipids were compared in cerebrum and cerebellum to establish the time, if any, at which their rate of accretion increases in a significant way. In the forebrain the rate of increase in concentration of these lipids accelerated at about the 32nd week of gestational age. Although there were too few postnatal cases to draw very firm conclusions, it seemed that the ganglioside concentration levelled off at about two months postnatal age and that the plasmalogen concentration reached a plateau between the 4th and the 6th postnatal months. In the cerebellum the concentration of gangliosides was clearly lower than that in the forebrain until about one year of age, the maximum rate of increase occurring between the last weeks of gestation and the second postnatal month. The plasmalogen concentration was somewhat higher in the cerebellum than in the forebrain but the concentration profile was similar to that followed by the gangliosides. In clear contrast with the concentration profiles in the cerebrum, in the cerebellum both lipids apparently continued to increase up to the second postnatal year. A mainly perinatal period of vulnerability is suggested for the forebrain, and a more prolonged one (probably until the second year of life) for the cerebellum.


A chemical study on the development of the human forebrain and cerebellum during the brain 'growth spurt' period. II. Phosphoglyceride fatty acids. The changes of the fatty acid composition of ethanolamine phosphoglycerides (EPG) and choline phosphoglycerides (CPG) that are associated with the maturation of the human forebrain and cerebellum have been studied. The increase in the n-3 fatty acids and the 22:4(n-6)/22:5(n-6) index in the forebrain during the second half of gestation was confirmed in the present series. In the cerebellum, on the other hand, these two parameters were higher than in the cerebrum and did not show any significant change throughout the period studied. The percentage of 18:1(n-9) changed in an opposite sense in the cerebrum and in the cerebellum, i.e. it decreased with gestational age in the cerebrum, whereas it increased markedly in the cerebellum. Consequently, the 18:0/18:1(n-9) index, which increased with maturation in the cerebrum, decreased in the cerebellum.


Preoperative evaluation of the patient with myocardial disease: clinical review. The patient with chronic disease of the myocardium must be evaluated perceptively as to the extent of abnormal function. The degree of reduction of cardiac reserve must be estimated and all efforts must be made to improve cardiac performance before operation; then a carefully managed anaesthetic is needed to assure a smooth operative and post-operative course.


The linear relation of cerebral blood flow to arterial oxygen saturation in hypoxic hypoxia induced with nitrous oxide or nitrogen. This study has demonstrated a linear relationship between cerebral blood flow and arterial oxygen saturation (Sao2) in the cerebral blood flow response in the hypoxic range of Pao2 values, which we believe is documented for the first time. The cerebral blood flow increased 1.02 per cent for each percentage decrease in Sao2 in hypoxia induced with nitrogen. Hypoxia induced with nitrous oxide increased cerebral blood flow at the rate of 2.09 per cent for each one per cent decrease in Sao2 which was dramatically more than the increase associated with hypoxia induced with nitrogen. Increased survival rates at lower Sao2 levels suggested that nitrous oxide also exerts a protective effect on dog myocardium. The mechanism by which hypoxia increases cerebral blood flow is unclear but the prevalent theory is one of neurogenic control which causes cerebral blood flow to increase as the intracellular hydrogen ion ([H+]I) increases in the progressive lactacidosis of hypoxia. The demonstration of a linear relationship between cerebral blood flow and Sao2 will provide the statistician, the researcher and the clinician with a useful tool for the neurophysiological assessment of cerebral blood flow.


Variability of the pulmonary vascular response to hypoxia and relation to gas exchange in dogs. Anaesthetized and mechanically ventilated dogs were subjected to five minutes of alveolar hypoxia with Fio2 ranging from 0.08 to 0.20 while pulmonary artery pressure (Ppa), pulmonary wedge pressure (Pwp) and cardiac output (Q) were measured. Hypoxia increased Ppa in all dogs whereas Pwp and Q did not change significantly. Thus pulmonary vascular resistance (PVR) increased by a mean for all dogs of 56 per cent. There was great variation in the absolute increase in Ppa and PVR between animals and these were not statistically correlated with arterial Po2, but highly significant and reproduceable inverse relationships were found for Ppa and PVR against arterial oxygen per cent saturation. The slopes of these responses varied between dogs from -0.013 to -0.144 for PVR and from -0.038 to -0.561 for Ppa. The alveolar-arterial oxygen gradient and the pulmonary shunt fraction were reduced in a similar way with decreasing arterial oxygen per cent saturation, but dead space/tidal volume ratio remained unchanged. Thus the slope of PVR response to hypoxia against arterial oxygen per cent saturation is unique for individual animals. This may reflect functional and likely structural adaptations of the pulmonary vascular smooth muscle.


Subanaesthetic halothane: its effect on regulation of ventilation and relevance to the recovery room. Ventilation and the ventilatory response to a steady-state of isocapnic hypoxaemia were measured in six healthy volunteers, both awake and while sedated with low doses of halothane (0.05 and 0.1 MAC). Halothane sedation markedly reduced ventilatory responses to sustained hypoxaemia, in a dose-related fashion. We estimated the length of time after anaesthesia that halothane 0.1 MAC would be present in patients in the recovery room. In five healthy patients who had halothane anaesthesia with a mean duration of one hour, halothane 0.1 MAC or more persisted for approximately one hour. We conclude that, during emergence from halothane anaesthesia, patients may have a significant impairment of the ventilatory response to hypoxaemia, which persists for some time even after regaining consciousness.


An assessment of operating room environment air contamination with nitrous oxide and halothane and some scavenging methods. This study was designed to assess operating room contamination with nitrous oxide and halothane when nitrous oxide 3 1/min and oxygen 2 1/min containing halothane one per cent were passed into a semiclosed circle absorber system from which the patients' lungs were ventilated with an Ohio ventimeter through a cuffed tracheal tube, with the exhaled gas vented to the floor. The normal room ventilation did not consistently maintain levels below the suggested amounts, which are nitrous oxide 25 ppm and halothane 2 ppm. Careful daily check for and correction of high and low pressure leaks combined with the use of a Foregger scavenging device (7--351--005) and continuous wall vacuum accepting approximately 22 1/min enabled geometric mean values below the suggested levels to be achieved. The Protection Filter Foregger 7--365--001 was only effective in removing halothane and only if the plastic holder provided was radically modified. Attention is drawn to the numerous factors influencing the magnitude of contamination, aspects of controlling it, and the necessity to cope with the problem of dealing with scavenged gases and vapours.


Serum cholinesterase activity following enflurane anaesthesia. Serum cholinesterase activity was determined in 30 female patients anaesthetized with enflurane for excision of lump in the breast. In all 30 patients biopsy of the tumor was negative for carcinoma. Blood samples were taken before induction of anaesthesia, at the end of the operation and 24 hours after the operation. Enzymatic determinations were performed by the Boerhinger cholinesterase kit. Enzyme levels were found sufficiently high at the end of the operation and returned to the preoperative levels 24 hours post-operatively.


Uneventful extradural analgesia after unrecognized dural perforation. A case is reported of an uneventful continuous extradural block in spite of unrecognized dural perforation. Although the perforation was recognized later during the block, the dura was probably pierced during the insertion of the catheter and only later the terminal hole came to lie in the subarachnoid space. Consequently both extradural and subarachnoid spread of the injected local anaesthetic occurred, preventing an extensive subarachnoid block.


Anaesthesia for ambulatory surgery.

Anaesthesia for ambulatory surgery. Most surgeons and hospital administrators now recognize that many surgical procedures can be done on an out-patient basis and anesthetists have been instrumental in developing and implementing the practice of short-stay surgery. It is our opinion that general anaesthesia using enflurane and Alfathesin will increase the safety of ambulatory surgery.


Major issues facing academic departments of anaesthesia in Canada. Two conditions must be present for a successful and strong department: (1) the utilization of operating rooms must be efficient, patient volume must be adequate and turnover rate be rapid, if the clinical income of academic staff is to be equal to that of nonacademic anaesthetists. This is really difficult with the surgical teaching that takes place in teaching hospital operating rooms. But if university hospital operating rooms are not comparably efficient, clinical incomes for academic anaesthetists are not competitive to private practice. (2) the university (i.e. the provincial government) must pay staff adequately for their academic time so that high-grade people will accept and stay in academic posts. Part-time staff deserve to be remunerated for their contributions, whatever the magnitude, or motivation to do it is minimal. If these basic criteria of financing a department are fulfilled, development of the academic functions will follow. Strong academic departments will be producing the capable anaesthetists to fulfill the manpower needs of our specialty in Canada. If not, more departments will atrophy and the manpower situation for anaesthetists in Canada will be in an acute crisis.


The current status of anaesthesia research in Canada. The Association of Canadian University Departments of Anaesthesia (A.C.U.D.A.) surveyed all 16 university departments during 1975/76/77, to obtain detailed and accurate information on manpower and research activities. The results of this survey have been summarized and show that only 0.68 per cent of federal MRC funds support anaesthesia research compared with 3.29 per cent of the U.S. federal N.I.G.M.S. funds. Notably 90 per cent of Canadian research funds are held in three departments. Canadian academic anaesthetists spend twice as much time in clinical service as their U.S. counterparts and there are 46 per cent fewer full-time faculty and 64 per cent fewer researchers in Canadian departments of Anaesthesia as compared to the U.S. A number of recommendations are made which will be acted upon as a basis for future planning.


Potato spindle tuber viroid: circular dichroism spectrum and physical chemical studies of its interaction with ethidium bromide. The presence and extent of base-paired secondary structure in the protein-free potato spindle tuber viroid RNA has been investigated by spectroscopic techniques. Dye-binding studies with the intercalating dye, ethidium bromide, indicate the presence of base-paired regions in the molecule. Variation of the induced chirality in the dye with the dye to RNA phosphate ratio indicate the presence of two base-paired regions in the molecule.


A rapid chromatographic method for the determination of omega-N-methylarginines in protein and muscle tissues. A rapid and sensitive chromatographic method is described for the complete separation and determination of NG-methylarginine, NG,NG-dimethlarginine, NG,N'G-dimethylarginine, and related compounds in protein and muscle tissue hydrolysates. This method is designed to be used both with conventional amino acid analyzers using a single column (10 X 0.9 cm) of Durrum type DC-6A resin at 51 degrees C, one buffer system (0.21 M sodium citrate pH 5.40), and one buffer flow rate (70 ml/h) and with the accelerated and fully automatic Beckman Spinco model 121M amino acid analyzer using a 10 X 0.28 cm microcolumn packed with the Beckman type AA-10 resin and operated with the same buffer at 7.9 ml/h. This procedure was successfully applied to the determination of omega-N-methylarginines in various animal tissues, and the presence of four as yet unidentified ninhydrin-positive compounds is reported.


Blood replacement in dogs by dextran-hemoglobin. Exchange transfusions in dogs were performed with a solution of either dextran or a covalent complex between dextran and human hemoglobin. Dogs transfused with dextran alone died when their hematocrit was lowered to 6-10%. Dogs transfused with dextran-hemoglobin complex, however, survived a reduction of their hematocrit to 2% or below. In the latter animals, the dextran-hemoglobin complex disappeared from the circulation with an average half-life of 2.4 days. Correcting for oxidation of the hemoglobin moiety to methemoglobin, the half-life of functional unoxidized dextran-hemoglobin in the circulation was 1.9 days. In compensation for the loss of dextran-hemoglobin, vigorous erythropoiesis was observed at a rate of close to 5% hematocrit per day over the first 2 days following the exchange transfusion. As a result, the total hemoglobin concentration in blood was maintained at 5-6% during this period, and the animals went on to complete recovery in room air without the need for further transfusion with dextran-hemoglobin.


A comparison of the in vitro metabolism of biphenyl and 4-chlorobiphenyl by rat liver microsomes. The comparative metabolism of the hydrocarbons, biphenyl and 4-chlorobiphenyl, was investigated using two different preparations of rat hepatic microsomes. The assay was designed to account for all the metabolic products which included the ether soluble lipophilic metabolites, low molecular weight conjugates, and macromolecular adducts, and to determine the effects of induction with Aroclor 1254 and 1248, two commercial polychlorinated biphenyl (PCB) preparations. 4-chlorobiphenyl was the more metabolically active substrate with the induced and control enzymes. In most metabolic fractions biphenyl was less inducible by the PCB's, with the exception of the 2-biphenylol metabolite which was induced ca. 18-fold. Preincubation of the microsomes with carcinogens did not enhance biphenyl 2-hydroxylation. Instead, a general inhibition of metabolic activity was observed for both biphenyl and 4-chlorobiphenyl substrates. Preincubation with phenobarbitone, a noncarcinogen, did not change the microsome-mediated metabolism of biphenyl or 4-chlorobiphenyl. The substitution of a single halogen atom on the biphenyl nucleus altered both the reactivity and pattern of metabolites for these substrates.


Chromium (III) thiamin diphosphate: a new probe of enzyme coenzyme--metal ion binding. Interaction with wheat germ pyruvate decarboxylase. Chromium (III) thiamin diphosphate (CrTDP) is a substitutionally inert complex which is a physical and kinetic probe of the binding mode of metal ion and coenzyme in thiamin diphosphate (TDP) dependent enzymes. CrTDP is prepared by reaction of aquated Cr (III) and TDP and purifed by ion-exchange chromatography. CrTDP binds to the apoenzyme of wheat germ pyruvate decarboxylase, giving an inactive holoenzyme. Chromium (III) ion binds to the apoenzyme in a manner that suggests, in light of the results with CrTDP, that there are two metal ion binding sites. Extension of the use of CrTDP to other enzymes can give specific information about binding and can introduce an active site reporter group


Effect of calcium on [125I]insulin binding to rat adipocytes. The specific binding of [125I]insulin to rat adipocytes was reduced in the absence of extracellular calcium. The addition of calcium to the calcium-free medium during incubation restored [125I]insulin binding towards normal. The specific binding of insulin was significantly increased with calcium concentrations as low as 0.5 mM and maximal binding occurred with 5 mM calcium. Scatchard analysis of the data suggests two major binding sites, one a high-affinity low-capacity site (Kd, 1.5 X 10(10) M-1) and the other a lower-affinity high-capacity site (Kd, 4.7 X 10(9) M-1). There was a 50% decrease in the number of high-affinity sites in absence of extracellular calcium. The dissociation curve of receptor-bound insulin was nonlinear both in the absence and presence of extracellular calcium suggesting receptor heterogeneity. The dissociation rate of receptor-bound insulin was greater when insulin was bound in the absence of extracellular calcium than in its presence. These results indicate that extracellular calcium, by increasing the number of high-affinity receptor sites, can alter the the ratio of high-affinity to low-affinity receptors for insulin in adipocytes.


Localization of fumarylacetoacetate fumarylhydrolase in rat liver. The intracellular location of fumarylacetoacetate fumarylhydrolase (EC 3.7.1.2) has been demonstrated in rat liver tissue. Two fractionation procedures involving homogenization and differential centrifugation were adopted. The first fractionation procedure isolated the nuclear fraction while the second gave the mitochondrial, microsomal, and soluble phase fractions. The hydrolase is localized in the soluble phase of the rat liver tissue. The enzyme also showed a high relative specific activity in the soluble phase fraction. Fractionation efficiency was checked by microscopic studies and by determining the distribution of a number of marker enzymes.


Microheterogeneity and sialic acid in human plasma angiotensinogens in various physiological states. Pooled or individual plasmas from normal men, women, pregnant women (third trimester), anephric women, and rat liver perfusates were used as sources of angiotensinogen. The plasmas were fractionated and desalted by Sephadex gel filtration, then subjected to isoelectric focusing in a pH 4 to 6 gradient on 40 X 4 cm slabs of polyacrylamide gel. The gels were cut transversely into 0.5-cm-wide strips, the pH measured, and their angiotensinogen concentrations determined by incubation with excess human renin and radioimmunoassay of the product, angiotensin I. This revealed several peaks of angiotensinogen concentration indicative of microheterogeneity in all cases. Contrary to other claims, the isoelectric pH profiles of angiotensinogens in the various physiological states were substantially alike. Major peaks were found at pH 4.75 to 4.85 and 4.9 to 5.0 and minor peaks at pH 4.5 to 4.7 and 5.0 to 5.2; this resolution was greater than that achieved with rat liver angiotensinogens. Incubation of human angiotensinogens with neuraminidase for 3 or 16 h raised their isoelectric pH by about 0.5 U, probably due to removal of sialic acid. Since microheterogeneity persisted after desialylation, it is probably determined by structural characteristics other than sialic acid composition.


Properties of a unique catalase isolated from Aspergillus niger. The properties of an unusual catalase isolated from Aspergillus niger are reported. The enzyme, which has a molecular weight of 323,000, consists of four subunits and is highly resistant to sodium azide.


Evaluation of staphylococcal thermonuclease (TNase) assay as a means of screening foods for growth of staphylococci and possible enterotoxin production. Samples of food, naturally and artificially contaminated with Staphylococcus aureus, were analyzed for enterotoxin and thermonuclease (TNase). With the exception of egg, all naturally contaminated foods with detectable amounts of staphylococcal enterotoxin were positive for TNase. The enzyme was also present in the majority of foods with over 1 million S. aureus cells per gram. In artificially contaminated foods. TNase was detected in all samples except egg, acidic foods in which growth of S. aureus was suppressed, and some samples of raw ground beef. Recovery of TNase was influenced by the type of food.


Interaction of atrazine with soil microorganisms: population changes and accumulation. A loam soil treated with atrazine at rates of 10, 30, and 100 microgram/g soil resulted in increased populations of actinomycetes, bacteria, and fungi over those in non-treated soil. The increases were in proportion to the amount of atrazine and persisted for at least 2 months. Living actinomycete amd fungal mycelia were incubated for 48 h in distilled water, nutrient broth, or soil containing 5 microgram/ml (g) of the herbicide. Actinomycete and fungal mycelia accumulated atrazine from water to concentrations up to 87-fold and 132-fold, respectively, over that in the ambient medium.  The maximum accumulation from soil by actinomycete mycelia was 26-fold and by fungal mycelia 13-fold. Fungi accumulated little or no atrazine from a nutrient medium whereas actinomycetes accumulated up to 13-fold. Dead mycelia usually did not accumulate atrazine in excess of the ambient concentration. Mycelium of Sclerotium rolfsii growing in a nutrient medium containing 20 microgram atrazine/ml accumulated 157 microgram/g wet weight after 8 days. Sclerotia subsequently produced after transfer of similar mats to soil contained 550 microgram/g wet weight.


Fungal exudates.

Fungal exudates. The exudates or liquid droplets on various structures of a number of fungi were examined. The droplets were enveloped in membranous material and were associated with actively growing mycelia, including fruiting structures. Osmium tetroxide vapour-fixed droplets of Claviceps purpurea, Myrothecium roridum, Sclerotinia sclerotiorum, Sclerotium rolfsii, and Thanathephorus cucumeris did not dry to a powder but remained intact as spheres when freeze-dried. Fractured spheres, examined with the scanning electron microscope, showed the presence of a membranous structure similar to that of rapidly frozen colloidal solutions with the ice crystals removed by sublimation. Locules or cavities within the freeze-dried droplets are thought to be due to the entrapment of air when droplets coalesce. Biochemical analyses of the exudates showed that acid phosphatase, beta-glucosidase, acid and alkaline protease. RNase polygalacturonase and cellulase enzymes as well as oxalic acid and ammonia were present.


The growth of Mycobacterium convolutum on solid n-alkane substrates: effect on cellular lipid composition. Growth rates and cell yields of Mycobacterium convolutum strain R22 grown on a wide range of both odd- and even-numbered carbon solid n-alkanes decreased as the substrate carbon number increased. Cellular lipid was 2.5 times higher following growth on the hydrocarbon substrates. The amount of polar lipid was found to be about half of the cellular lipid in docosane through octacosane-grown cells. Phosphatidylethanolamine represented about 50% of the polar phospholipid in hydrocarbon- and nonhydrocarbon-grown cells. Phosphatidylserine, diphosphatidylglycerol, and phosphatidic acid were also found. The amount of phosphatidylserine was higher (10-14%) in docosane through octacosane-grown cells. There was a correspondingly smaller amount of diphosphatidylglycerol. These lipid changes may be associated with the assimilation of the hydrophobic substrates. The failure to detect label in lipids of solid n-alkane-grown cells incubated in the presence of [14C]acetate suggested that de novo synthesis of fatty acids did not occur to an appreciable extent. Transport of [14C]acetate and incorporation into cellular protein was not inhibited. Polar lipid fatty acid analyses indicated there was no direct incorporation of the oxidized substrate. The results suggest that beta-oxidation and a subterminal oxidative cleavage appear to be the major catabolic routes providing fatty acids, which are then incorporated into lipid.


Relative microbial activity and bacterial concentrations in water and sediment samples taken in the Beaufort Sea. A total of 91 water, 8 ice, and 50 sediment samples taken from the southwestern Beaufort Sea were analyzed for relative microbial activity, bacterial cell concentrations, and percentage of respiration (mineralization). These samples were taken during three field-study periods (August to September 1975; April 1976; and August 1976). Both the relative microbial activity and the cell concentrations in water and sediment samples were lower during the April (winter) sampling period than in the August-September (summer) studies. The percentage of respiration of labeled glutamic acid was higher in the winter water samples than in the summer samples. The water samples showed higher percentage of respiration values than did sediment samples. The average maximimum potential rate of glutamic acid uptake was as high or higher than those observed in studies made in more temperate waters. Samples of melted sea ice showed levels of relative microbial activity that were about the same as that found in the associated seawater. When 1:1 mixtures of melted ice and seawater were analyzed for altered microbial activity, little effect could be detected.


The production of extracellular amino acids by rumen bacteria. One hundred and sixteen freshly isolated rumen bacteria and 10 laboratory strains were studied for the production and excretion of free amino acids during growth in a basal medium containing glucose, cellobiose, and soluble starch as the energy sources, (NH4)2SO4 as the prime nitrogen source, volatile fatty acids, hemin, vitamins, Na2CO3, and cysteine as the reducing agent. Amino acid analyses of 48-h culture fluids of the isolates indicated the presence of alanine, glutamic acid, valine, aspartic acid, glycine, serine, lysine, methionine, leucine, isoleucine, threonine, histidine, arginine, phenylalanine, and tyrosine. Most isolates excreted some free amino acids. Alanine, glutamic acid, valine, aspartic acid, and glycine were found in the greatest concentrations with some isolates accumulating between 50 and 295 microgram/ml of one or more of these compounds. Concentrations of the remaining amino acids rarely exceeded 20 microgram/ml of culture fluid. Growth studies demonstrated that the amino acids were excreted during active growth of the bacteria and ceased shortly after growth became limited.


Endotoxic activity of cell-free rumen fluid from cattle fed hay or grain. The cell-free rumen fluid from cattle fed hay or grain exhibited the following biological characteristics which strongly suggest the presence of endotoxin or a toxic principle similar to endotoxin of gram-negative bacteria: proved lethal to mice when injected with actinomycin D; proved extremely lethal to chick embryo; induced biphasic pyogenic response in rabbits; enhanced susceptibility to bacterial infection in mice; evoked positive epinephrine skin reaction in rabbits and phenol-water or aqueous ether proved lethal to mice and chick embryos. A quantitative difference in concentrations of endotoxin was observed on LD50 in mice and chick embryos and response to the epinephrine skin test in rabbits. Cell-free rumen fluid of grain-fed cattle contained at least twice as much endotoxin as that of hay-fed cattle. Endotoxin in cell-free rumen fluid and in higher concentration in cattle fed grain than in those fed hay support the hypothesis that rumen bacterial endotoxins may participate in the pathogenesis of diseases associated with high grain feeding such as lactic acidosis and the sudden-death syndrome.


Studies on bacteroid size and nucleic acid content of alfalfa bacteroids fractionated by velocity sedimentation. A velocity sedimentation procedure was described to fractionate bacteroids of alfalfa nodules into four subpopulations. Bacteroids in these subpopulations were different in size and nucleic acid content as determined by microscopy and flow-microfluorometry (FMF). The slowest-sedimenting bacteroids (fraction I) were small and resembled free-living Rhizobium meliloti both in size and nucleic acid content. The fastest-sedimenting bacteroids (fraction IV) were 2 to 3 times longer and contained 3 to 4 times more nucleic acid than the small bacteroids in fraction I and free-living R. meliloti. A positive correlation was established between bacteroid size and relative nucleic acid content of bacteroids in alfalfa nodules.


"The 'impossible' child," the difficult child, and other assorted delinquents: etiology, characteristics and incidence. The evidence is very strong and persuasive that if serious juvenile delinquents are identified as having reached Level 4, a differential treatment program can be identified for them which will increase their chances of avoiding further law violation behaviour. Looking at both I-level growth and further delinquent behaviour, less is known about how to bring about change in the Power-oriented youths than in other I-level groups. This group is perhaps the most deserving of the label "Impossible Child." Continuing attempts to treat the Power-oriented offender in experimental programs is extremely important because this group includes the most serious offenders (the most felony-type crimes and the most assaultive crimes). More research is also needed on the treatment of the Passive Conformist group. We need to know more about the reasons for the short-term success and the long-term failure. Although this subtype does not commit as serious crimes as the other I3 subtypes, there is some evidence that there is a persistence to the offence behaviour which continues into adulthood, producing highly recidivistic individuals.


Gray scale ultrasonography in the evaluation of metastatic melanoma. One hundred and eight ultrasonographic examinations of the abdomen and pelvis in 89 patients with malignant melanoma were reviewed. Forty of these patients were found to have metastatic disease demonstrated by this new diagnostic modality. The varied ultrasonographic manifestations of melanoma are illustrated. Its use in staging and evaluating therapy is stressed.


Thyroid abnormalities following neck irradiation for Hodgkin's disease. Thyroid function and scans were evaluated in fifty disease-free patients 2 to 16 years after receiving neck irradiation for the treatment of Hodgkin's disease. Twenty-five of 50 patients had abnormal thyroid studies: eight were hypothyroid, two were hypothyroid and had abnormal scans, and fifteen had abnormal scans. Of the 15 patients with abnormal scans, one had an isolated elevation of TSH (thyroid stimulating hormone) and one developed exophthalmos. These data, obtained within a relatively short follow-up period, indicate that morphologic and functional abnormalities of the thyroid gland are not uncommon in patients who have received irradiation to the thyroid gland in the course of treatment for Hodgkin's disease. There is need for continuous reevaluation of the thyroid status in such patients.


Single dose X-irradiation and concomitant hyperthermia on a murine fibrosarcoma. The objectives of this study were to quantitate the effects of local tumor hyperthermia (LTH) and concomitant x-irradiation (RAD) on a moderately radioresistant murine fibrosarcoma in situ. Comparisons were made to the combined treatment response on the Ridgway osteogenic sarcoma, a radio sensitive tumor previously used in this laboratory and to establish the Meth-A fibrosarcoma as a model system for combined modality studies. 1.0 cm3 tumors were exposed to single doses of RAD ranging from 0.5-3.5 krad alone or 0.5-2.3 krad in combination with LTH (water bath at 43.1 +/- .05 C for 20 minutes) applied immediately postirradiation. LTH significantly enhanced the action of radiation as measured by tumor volume analysis, mean survival time and cures. The ratio of radiation doses vs. RAD + LTH required to produce an equivalent response ranged from 1.4 to 2.5 depending upon the endpoints evaluated. These findings are consistent with single dose studies on the radiosensitive Ridgway osteogenic sarcoma and suggest that the tumoricidal effectiveness of combination radiation and hyperthermia cannot be predicted on the basis of the radiation alone responsiveness of tumor.


The significance of fractionation regimens in radiation and combined hyperthermia using a murine fibrosarcoma. The significance of time-dose relationships in the use of local tumor hyperthermia (LTH) when combined with radiation (RAD) was studied in a murine fibrosarcoma. RAD, either alone or combined with LTH, was delivered in four equal fractions (total doses, 1.8 to 4.2 krad) separated by 1 to 4 days. LTH (43.1 C +/- .05 C for 15 minutes, water bath) was applied immediately after RAD. In this tumor system, RAD was most effective when delivered every 2nd or 3rd day, by a factor of 1.25 over the response achieved when the four fractions were delivered every 1 or 4 days. At all levels studied, RAD + LTH produced a superior tumor response compared to RAD alone. The ratio of the RAD + LTH/RAD doses to achieve an isobiological response ranged from 1.7 to 2.5. Most significant was the finding that the RAD + LTH treatment response was independent of the fractionation scheme used and more dependent on the total RAD dose delivered.


Radiation therapy in the treatment of superior vena caval obstruction. The obstruction of the superior vena cava by tumor is recognized as an acute or subacute oncologic emergency. Rapid high-dose irradiation to the mediastinum is shown to be effective therapy for a superior vena caval obstruction. In our series 35 patients have been treated with rapid high-dose irradiation between January 1971 and July 1977. The present treatment consists of 400 rad given in a daily midplane dose for 3 days, and then slowing to 150 rad midplane per day to a total dose of 3000 rad over 15 fractions. There have been no instances of exacerbations of symptoms or severe complications from this treatment. There have been 2 failures of the 35 patients treated with this regimen. Both were shown at autopsy to have massive thrombi obstructing the superior vena cava. Six years of experience using this regimen has proven to be a safe and effective means of treatment for superior vena caval obstruction.


Specific immunotherapy of established visceral micrometastases by BCG-tumor cell vaccine alone or as an adjunct to surgery. A vaccine of Bacillus Calmette-Guérin (BCG) admixed with tumor cells induced systemic immunity and had a therapeutic effect on subclinical, disseminated micrometastases. Inbred strain 2 guinea pigs given intravenous injections of either 10(4), 10(5) or 10(6) syngeneic L10 hepatocarcinoma cells were vaccinated after metastatic foci were established in the lung parenchyma. The studies demonstrate that under defined conditions of vaccine preparation and regimen, nontumorigenic preparations of BCG and tumor cells can cure the majority of animals of otherwise lethal visceral metastases. Histopathologically it was determined that immunization with these vaccines prevented the progressive growth of pulmonary micrometastatic foci approximately 0.1 mm in diameter. However, in this micrometastasis therapy model, the number of metastatic tumor foci is a major limitation in the efficacy of vaccine therapy. No protection against L10 tumor was achieved when antigenically distinct but syngeneic L1 hepatocarcinoma was used in the vaccine, suggesting that this is a tumor-specific immunotherapeutic procedure. This BCG-L10 tumor vaccine was also effective in curing guinea pigs of minimal disseminated tumor burden when administered after surgery of an established skin tumor and draining lymph node.


Association of Kaposi's sarcoma and prior immunosuppressive therapy: a 5-year material of Kaposi's sarcoma in Norway. A retrospective study of 53 cases with a histological diagnosis of Kaposi's sarcoma (KS) reported to The Cancer Registry of Norway during a five year period is presented. Four cases were excluded from the material because further information contradicted the diagnosis of KS. Of the remaining 49 cases, information of treatment received before the development of KS was obtained in 41 (83.7%). Six (14.6%) of the 41 patients developed KS during systemic treatment with corticosteroids, two of the 6 cases also used azathioprine. None of the patients had undergone renal transplantation. One additional patient had received radiotherapy for a malignant lymphoma prior to the development of KS. In a control group of 242 consecutive patients hospitalized because of basal cell carcinoma of the skin, none had used systemic corticosteroid or cytotoxic drugs. Case reports of the 7 patients with KS developing after immunopressive therapy are presented.


Intralesional injection of the methanol extraction residue of Bacillus Calmette-Guerin (MER) into cutaneous metastases of malignant melanoma. Twenty-two patients with cutaneous metastases of malignant melanoma were treated with intralesional injections of the methanol extraction residue of bacillus Calmette-Guerin (MER). The local reaction consisted of erythema and pustule formation followed by ulceration and tumor necrosis. Side effects included fever, chills, headache and malaise in the majority of patients; nausea, vomiting, cyanosis and hypotension occurred infrequently. Hypersensitivity reactions were not observed. Temporary abnormalities in liver function were seen in 11 of 19 patients tested. Reversible lymphopenia and thrombocytopenia developed in 7 of 17 and 7 of 18 patients, respectively. Immune function, as measured by skin tests for delayed hypersensitivity and the in vitro response of isolated lymphocytes to mitogens and microbial antigens, was not influenced by treatment with MER. Transient increases were observed in total hemolytic complement, complement components and the reduction of nitroblue-tetrazolium by neutrophils. Eight of eighteen evaluable patients showed a complete disappearance of all injected lesions. We conclude that intratumoral injection of MER is effective treatment for cutaneous metastases of malignant melanoma, with a complete response rate comparable to that observed after intralesional injection of BCG.


Endolymphatic stromal myosis: unique presentation and ultrastructural study. We describe a case of endolymphatic stromal myosis (ESM) in which the lesion presented uniquely as recurrent frond-like cervical polyps. The lesion behaved aggressively, spreading to the peritoneum. Ultrastructurally, this tumor closely resembled proliferative phase endometrial stroma, in accord with the theory that ESM is a low-grade stromal sarcoma. The ultrastructural correlations may aid in developing future therapy, including progestagen administration for these unusual tumors.


Ultrastructure of the goblet cell type of adenocarcinoid of the appendix. Two examples of the goblet cell form of adenocarcinoid were studied with electron microscopy. Mucin was present in the form of large and small vacuoles or small patches of denser mucin granules. A small population of cells had a complement of organelles similar to the more obvious goblet cells but contained little or no identifiable product. Enterochromaffin cells were similar to those seen in normal midgut and in conventional carcinoids of midgut derivation. Enterochromaffin granules and mucin were not mixed with one another. Cell types transitional between mucinous and enterochromaffin were not found. Although study of plasmalemmae was hindered to a degree by artifacts due to tengential sectioning and, possibly, insufficiently rapid fixation, we favor the interpretation that the goblet cell type of adenocarcinoid is composed of two separate classes of cells.


Axillary lymph-node and breast carcinoma interrelations in organ culture. When Stage I and II cancers of the breast and their axillary lymph nodes were grown in the same tissue culture, a phenomenon of lymphocytic migration from the nodal explants to the tumor explants was observed. The lymphocytes infiltrated in and around the tumor nodules with cytotoxic effects; concomitantly, there was lymphocytic depletion in the nodal explants. The lymphocyte migration was particularly apparent when the axillary lymph nodes showed hyperplasia of the paracortical area and/or sinus histiocytosis. No correlation was found between the migration and the histologic type of disease or the degree of malignancy, but a strong correlation was observed between 1) the migration and the presence or absence of metastases in the explanted lymph nodes, and 2) the extent of the metastatic involvement in vivo. The lymphocyte migration was present only in the patient either uninvolved lymph nodes or only a small number (1-3) of metastatic nodes.


True hermaphrodite with multiple gonadal neoplasms: report of a case with cytogenetic study. On account of the functioning bigonads, true hermaphrodites show a wide range of morphological anomalies. The patient under discussion had an ovary on one side and an ovotestis on the other side; both of them functioned to some extent to produce a rudimentary uterus, fallopian tubes and a few Woolfian remnants. The additional point of interest in this report was the presence of three gonadal neoplasms-namely Yolk Sac Carcinoma gonadoblastoma and seminoma. While the ovary on one side was replaced by yolk sac carcinoma, the ovotestis was partly destroyed by the other two. The cytogenetic study revealed a hypodiploid number and a mosaic sex chromosomal pattern. The karyotypic abnormalities noticed in the patient are also reviewed.


A human plasma component that binds benzo(a)pyrene. A component capable of binding benzo(a)pyrene was measured in plasma from cigarette smokers and nonsmokers. This plasma fraction was found to have a high specificity of binding to benzo(a)pyrene, bound benzanthracene competitively with benzo(a)pyrene, and was positively correlated (r = 0.861, p less than 0.001) with the capacity of the individual subject's lymphocytes to be induced for AHH activity in culture. An inverse correlation (r = -0.957, p less than 0.001) between the presence of the plasma component in lung cancer patients and the capacity of lung cancer patients' lymphocytes to be induced in culture is unexplained at this time. A benzo(a)pyrene-binding fraction was not found in induced or uninduced cultured lymphocytes from smokers or nonsmokers, or in homogenates of lung excisional tissue from smokers with or without primary lung cancer.


Sarcoma arising in oligodendroglioma of the brain: a case with intramedullary and subarachnoid spinal metastases. A case of sarcoma arising in a left temporal oligodendroglioma is described in a 39-year-old male. The postoperative evolution was marked by a paraplegia linked to medullary and subarachnoid metastases. Then a relapse of the cerebral tumor occurred, and the patient died 6 months after the onset of the symptoms. Histologically, the mesenchymal component appeared fibrosarcomatous and angiosarcomatous. Those facts argue in favor of the histogenic hypotheses put forward in exceptional cases previously described, according to which the sarcoma develops from vascular elements of the glioma.


Bladder carcinoma associated with ectopic production of gonadotropin. A rare incidence of a primary gonadotropin-producing bladder carcinoma in which gynecomastia appeared in the terminal stage was encountered in a 76-year-old Japanese male. There was a good probability that the symptoms of hormonal activity were due to chorionic gonadotropin (hCG), since its whole molecule, beta- and alpha-subunits were detected by radioimmunoassay in the blood, urine, and the tissue from the malignant neoplasm, and the plasma and urine estrogens were elevated. Recent papers concerning the synthesis of hCG-like material by neoplastic cells are reviewed and the implication of the measurement of beta- and alpha-subunits of hCG in various neoplastic diseases are discussed. Other characteristic profiles of plasma hormonal levels are also discussed in this case.


Adenocarcinoid, a mucin-producing carcinoid tumor of the appendix: a study of 39 cases. Adenocarcinoid is a form of appendiceal carcinoid possessing features of both carcinoid and adenocarcinoma. There are two histologic types. Thirty patients had the goblet cell type, characterized by nests of large mucin-distended cells. Nine patients had the tubular type, characterized by small glandular structures lined by uniform cells. Despite abundant mucin and a goblet cell or acinar-like arrangement, a closer relationship to carcinoid than to adenocarcinoma is suggested by a concentration of tumor elements below the crypts of Lieberkuhn, a lack of evidence of neoplastic transformation of the appendiceal mucosa, and the demonstration of argentaffin or argyrophil granules in 88% of the lesions. Six tumors, all of the goblet cell type, metastasized and resulted in the death of the patients. One of the tumours that metastasized had a prominent tubular component. Most adenocarcinoids can be adequately treated by appendectomy, but hemicolectomy is recommended for those tumors showing atypical foci, a high mitotic count, or spread beyond the appendix.


Carcinoembryonic antigen and ferritin in patients with lung cancer before and during therapy. Carcinoembryonic antigen (CEA) levels were determined in 114 patients with confirmed lung cancer at the time of diagnosis using the CEA Ire-Sorin radioimmunoassay. Elevated CEA values were found in 47%. Most of the patients with high CEA levels had clinically detectable metastases. Ferritin was detectable by the Laurell-electrophoresis in the serum of 58 out of 81 (72%) of the patients with confirmed lung cancer at the time of diagnosis. Ferritin levels were significantly higher in patients with metastases. Serial measurements of CEA and ferritin during radio- and chemotherapy showed that the assay may be useful to evaluate the effects of therapy. Because of some false negative results both CEA and ferritin determinations should be used only in context with other clinical and laboratory findings.


Identification of breast cancer patients with high risk of early recurrence after radical mastectomy. II. Clinical and pathological correlations. A report of the Primary Therapy of Breast Cancer Study Group. A prospective study of factors that might be helpful in predicting recurrence of breast cancer during the 2 years after radical mastectomy has been completed in 381 women by the Cooperative Breast Cancer Group (National Cancer Institute). Identification of clinical factors which might be associated with such recurrence has been achieved. A multivariate analysis of the data was oriented toward the identification of clinical factors other than lymph node status that might be simultaneously used to predict recurrence because of the current trend of cancer therapy toward more limited surgery. Degree of differentiation of the tumor, blood vessel invasion, patient age and tumor size were identified as important predictors of recurrence for premenopausal patients and tumor size was identified as important for postmenopausal patients. The addition of axillary lymph node status to these factors, however, made a significant improvement in the prediction equation for both pre- and postmenopausal patients. Studies of this type are of particular value to understand further the biology of breast cancer which is necessary to develop rational primary and adjuvant treatment strategies.


Metabolic epidemiology of large bowel cancer: fecal bulk and constituents of high-risk North American and low-risk Finnish population. The dietary pattern and fecal constituents of two populations with distinct risk for the development of colon cancer, a high-risk population in New York Metropolitan area and a low-risk population in rural Kuopio, were studied. The average daily intake of dietary fat and protein was the same in the two groups, but the sources of fat were different, a greater portion coming from dairy products in rural Kuopio and from meat in the New York Metropolitan area. Not only the frequency of bowel movements was higher, but also the daily total stool output as well as the fecal fiber excretion were greater in Kuopio compared with New York due to high dietary intake in rural Kuopio of cereal products rich in fiber. The concentration of fecal secondary bile acids and bacterial beta-glucuronidase activity was lower in rural Kuopio, but the total daily excretion of these constituents was the same in two populations. The daily fecal excretion of bacterial nuclear dehydrogenase activity and of neutral sterols was higher in rural Kuopio, and the concentration of these constituents was the same in the two groups. The high daily excretion of cholesterol metabolites in Kuopio might be due to high dietary intake of dairy products. The data suggest that one of the factors contributing to the low-risk of large bowel cancer in Finland, in spite of high dietary intake of fat, appears to be the fact that a high dietary fiber leads to an increase in stool bulk, thus diluting bile acids, which have promoting activity.


Polyps of the large intestine in Northern Norway. In unselected autopsies on 171 males and 109 females over 20 years of age in Tromsø, Northern Norway, adenomas of the colon and rectum were present in 40% of the males and 33% of the females. The frequency of adenomas increased with age in both sexes. Hyperplastic polyps were found in 20% of all cases with no variation in frequency with sex or age. Before the age of 65 most adenomas were located in the distal half of the large intestine in both sexes. After 65 years in males and 75 years in females the predilection site was shifted to the proximal half of the large intestine. The average size and grade of atypia of adenomas increased with age, but no significant difference in grade of atypia was found between the proximal and distal halves of the large intestine. The occurrence of adenomas was not associated with any of the common chronic diseases thought to be related to a western-style diet or to malignant or benign neoplasms elsewhere in the body. A significant association was found between the occurrence of adenoma and high body weight. The findings support the role of adenomas as precursors of colorectal carcinoma.


Colonoscopic biopsy and cytology in the diagnosis of colon cancer. Colonoscopy has revolutionalized the approach to the diagnosis and management of patients with colorectal neoplasia. When malignant-appearing lesions are visualized by colonoscopy, a variety of diagnostic techniques are currently available for the assessment of the nature of the lesion including biopsy, brush cytology, and lavage cytology. Comparison of results for biopsy alone with biopsy plus either or both cytologic techniques showed a positive yield of 60% for biopsy alone; 76% for biopsy and lavage; 89% for biopsy, brush, and lavage. When the cancers were divided into infiltrative and exophytic lesions the positive yield for biopsy alone was 33% for infiltrative, cancer, and 71% for exophytic cancer; for biopsy and lavage cytology, 44% for infiltrative cancer, and 94% for exophytic cancer; for biopsy and brush cytology, 78% for infiltrative, and 94% for exophytic cancer; and for biopsy, brush, and lavage cytology, 83% for infiltrative cancer, and 92% for exophytic cancer. The use of brush cytology improved the yield of tissue diagnosis considerably when added to the biopsy technique. Lavage cytology did not seem to increase significantly the diagnostic yield. The diagnostic yield of the various techniques was related not only to the specific combination of techniques used, but also to the gross tumor pattern.


Carcinoma of the floor of the mouth.

Carcinoma of the floor of the mouth. Records of all patients with carcinoma of the floor of mouth seen at the Indiana School of Medicine University Hospital between March 1965 and March 1975 were reviewed. 126 patients with these tumors were seen during this ten-year period. This study included patient's age, sex, race, family history for cancer, type of treatment received, treatment results and complications. The complications and duration of hospital stay were also studied. 18 of the 126 patients received surgery alone and 108 patients received either preoperative, postoperative or curative radiation therapy. Long-term disease control rates in the early stages of carcinoma of the floor of mouth treated by radiation therapy alone were equivalent or superior to results obtained by surgery alone and patients experienced considerably less complications. In more advanced cases, radiation therapy played an important adjunctive and palliative role.


Malignant transformation of cryopreserved early passage syrian golden hamster cells by 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide. 2-(2-Furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2) induced the malignant transformation of secondary cultures of Syrian golden hamster embryo cells prepared from cryopreserved primary cells. Transformed cells grew in semi-solid agar medium and formed sarcomas when inoculated subcutaneously into non-immunosuppressed suckling hamsters.


Comparison of benzo[a]pyrene metabolism by human peripheral blood lymphocytes and monocytes. Here we report a more sensitive high-pressure liquid chromatographic (HPLC) assay of benzo[a]pyrene (BP) metabolites elaborated by human monocytes and lymphocytes using only 10 X 10(6) cells for HPLC analysis. The major metabolites formed by both lymphocytes and monocytes were 3-hydroxy-BP, 9-hydroxy-BP and quinones. BP-dihydrodiols were also found by HPLC analysis, the major one being BP-7,8-dihydrodiol for both cell types. In addition, a peak slightly more polar than 9,10-dihydrodiol was formed by lymphocytes. The 7,8-diol peak was eliminated by the addition of 1,1,1-trichloropropene oxide to the incubation mixture. The presence of alpha-napthoflavone resulted in an overall decrease in metabolite production.


The binding of 14C-labelled acetylaminofluorene to nuclear proteins and DNA. The covalent binding of acetylaminofluorene (AAF) to rat liver nuclear protein fractions and DNA has been examined after 2 doses of 14C-labelled carcinogen (100 micronCi intraperitoneally) given at 24-h intervals. Most of the AAF (approx. 80%) was bound to the NHP which had the highest specific activity, relatively small amounts were bound to histones, residual proteins and DNA. After 2 weeks the label was more evenly distributed but little remained bound to the DNA. Pre-feeding rats with phenobarbitone (1 mg/ml in drinking water) reduced binding in all fractions. Pre-feeding rats with sodium sulphate (1 mg/ml in drinking water) also slightly lowered the binding to all fractions. Isoelectric focusing analysis (in polyacrylamide gels) of the NHP showed a complex binding pattern with no one component predominantly labelled, variations in the pattern were found after feeding sulphate and phenobarbitone. Normal histone components in the histone fraction (acid extract) contained only 25% of the labelled carcinogen present.


Stimulation of the synthesis of the H1 and H3 histone fractions of mouse epidermis by 12-O-tetradecanoylphorbol-13-acetate. Histones from mouse epidermis were fractionated chemically by the method of Johns, and the 5 resulting fractions were subjected to acrylamide gel electrophoresis at pH 3.2 using 15% gels. Their migration sequence was observed to be the same as that reported for calf thymus histones. Total epidermal histone from mice treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) and a pulse of [3H]lysine was also subjected to acrylamide gel electrophoresis and 5 separate bands were detected. Large increases in the incorporation of [3H]lysine into the H1 and H3 histones were observed when compared to histone bands from control mice, with smaller but significant increases of incorporation occurring into the H2B, H2A, and H4 fractions.


Concanavalin A agglutination of bladder cells of rats treated with bladder carcinogens; a rapid new test to detect bladder carcinogens. N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN), which is a potent, specific bladder carcinogen in rats, and its related compounds were orally administered to rats for 1 week. The bladder cells were isolated by the treatment with EDTA and sonication and they were subjected to agglutination assay by concanavalin A (Con A). Bladder cells obtained from rats treated with BBN, N-butyl-N-(3-carboxypropyl)nitrosamine (BCPN), N-ethyl-N-(4-hydroxybutyl)nitrosamine (EHBN) and N-butyl-N-(2-hydroxyethyl)nitrosamine (BHEN) were agglutination-positive, while those cells treated with N-tert-butyl-N-(4-hydroxybutyl)-nitrosamine (t-BBH) and N-butyl-N-(3-hydroxypropyl)nitrosamine (BHPN) were negative. Results obtained by this method were highly correlated with the known carcinogenicity of BBN and its analogues. Therefore, this method could be used for rapid screening of bladder carcinogens.


Therapeutic consequences of antitumour drug interactions: methotrexate and 5-fluorouracil in the chemotherapy of C3H mice with transplanted mammary adenocarcinoma. Methotrexate and 5-fluorouracil at fixed dosages of 1.5 mg/kg and 50 mg/kg, respectively, were employed in combination chemotherapy for transplanted mammary adenocarcinoma in C3H mice. Tumour regression analyses demonstrated the importance of drug sequence and scheduling. Significantly, a drug synergism and antagonism was exhibited under certain conditions. Results indicated a superior therapeutic efficacy when methotrexate was administered 6-12 h before 5-fluorouracil; the reverse proved antagonistic. However, at longer intervals a commutative additivity was apparent. Judicious scheduling of methotrexate and 5-fluorouracil would seem appropriate, as both drugs are widely employed in the current management of neoplasms, such as breast carcinoma.


Lifespan carcinogenicity studies with hexachlorophene in mice and rats. Hexachlorophene (HCP) was tested for carcinogenicity in lifetime feeding studies in male Sprague--Dawley rats that were fed on a protein- and vitamin-deficient diet and in C57B1 and XVII/G mice that received the chemical incorporated into a complete diet. After 2 years, no significant carcinogenic effects were observed in these animals. In XVII/G mice injected subcutaneously at birth or receiving HCP via the mother's milk, the incidence of tumors was not statistically increased. No carcinogenic effect was observed in (C57B1 X C3H) F1 hybrid mice that received HCP transplacentally.


Carcinogenic effect of N-nitrosobis(2-hydroxypropyl)-amine in rabbits. The carcinogenic effect of N-nitrosobis(2-hydroxypropyl)amine (BHP) was studied in rabbits. The animals received the compound in drinking water in concentrations of 1000 ppm for 25 weeks and the survivors were sacrificed 60 weeks after the treatment was begun. Whereas no tumors were found in controls, 3 out of 6 treated rabbits developed hapatic lesions (hepatocellular adenoma, hemangioma, and angiosarcoma), and 3 animals developed lung tumors (adenoma and hemangioma); no pancreatic neoplasms were observed.


Ritalin, Benzedrine and Dexedrine do not transform F 1706 rat cells. Ritalin (methylphenidate hydrochloride), Benzedrine (amphetamine sulfate), and Dexedrine (dextroamphetamine sulfate) were examined for oncogenicity using a serial line of Fischer rat embryo cells, which was previously shown to be a sensitive and accurate indicator of chemicals carcinogenic for rodents. At the concentrations tested, none of the 3 drugs induced transformation in Fischer rat embryo cells.


Low-molecular weight nucleic acids in the blood of rats with Zajdela's hepatoma. The phenolic method was employed to isolate 2--3 microgram/ml of nucleic acid from rat blood serum. Electrophoresis in an 8% polyacrylamide gel revealed 3 fractions corresponding to 7--9S RNA, and a highly polymer material retained at the beginning of the gel. The blood of the rats with transplanted Zajdela's hepatoma, pre-treated with CCl4, contained an immunodepressive nucleic factor (5S DNA) undetected in healthy animals. This factor may contribute to the suppression of antitumor immunity. The nucleic factor was not detected following treatment with CCl4 alone. It is suggested that the liver hinders nucleic factor transfer from the tumor environs into the blood.


Immunopotentiation and tumor inhibition with levamisole. The immunopotentiating drug, levamisole, was found to augment human lymphocyte responses to vegetable mitogens in vitro. However, the effect was dose-dependent and at high concentrations suppression rather than augmentation of the immune response was observed. In view of the potential widespread use of the drug in the treatment of human cancer, a study was undertaken in rats bearing breast cancer to determine whether a similar dose-response effect would be observed in terms of tumor growth. Tumor inhibition was found to be dose-dependent, and at high doses tumor inhibition did not occur while at lower doses the growth of the cancer was inhibited. The rate of tumor growth correlated well with the responses of splenic lymphocytes to vegetable mitogens. In a later study, it was not possible to show that levamisole had any effect in boosting immune responses of patients undergoing surgery. It is concluded that levamisole is capable of increasing immune responses in man in vitro, and that the drug is able to inhibit the growth of breast cancer in the rat. However, these effects are dose-dependent, and clinical trials of the use of levamisole in patients with cancer should include careful immune monitoring to be sure that augmentation rather than suppression of the immune response is being achieved.


Effects of Bacillus Calmette-Guérin and levamisole on immune responses in young adult and age-immunodepressed mice. The immunomodulating effects of bacillus Calmette-Guérin (BCG) and levamisole were tested in young adult mice after a single administration and in 12-month-old mice after continuous administration. In young mice, BCG was shown to activate macrophages, to potentiate antibody responses and delayed hypersensitivity reaction, to increase antibody-dependent cell-mediated cytotoxicity, and to induce nonspecific suppressor cells. Levamisole was not able to modify any of these immune responses in young mice. The action of these adjuvants differed markedly when tested in age-immunodepressed mice. BCG was found to be strongly immunosuppressive on antibody formation and induced suppressor cell activity in the spleen. Moreover, the survival of immunodepressed mice continuously treated with BCG was shortened in comparison to untreated aged mice. In contrast, levamisole acted as an immunorestoring agent because it strongly stimulated the antibody response compared to aged controls and did not induce suppressor cell population. The survival of levamisole-treated mice was prolonged when compared to untreated aged mice. When the surviving mice were killed and autopsied at the age of 24 months, the incidence of spontaneous tumors was significantly lower in the group of mice treated by levamisole.


Response of disseminated breast cancer to combined modality treatment with chemotherapy and levamisole with or without Bacillus Calmette-Guérin. One hundred and fourteen evaluable patients with measurable metastatic breast cancer were treated with a combination chemoimmunotherapy program (5-fluorouracil, adriamycin, and cyclophosphamide [FAC-levamisole [LSM]). An additional 117 patients with similar characteristics were treated with the same program with the addition of bacillus Calmette-Guérin (BCG) by scarification (FAC-BCG-LMS). The results were compared to those observed in 44 evaluable patients treated with FAC chemotherapy alone. Overall remission rates were identical in the three studies (72%, 73%, and 72%). The median duration of remission was 9 months for FAC, 13 months for FAC-LMS (P = 0.07), and 14 months for FAC-BCG-LMS (P = 0.05). The median duration of survival was 17 months for FAC and 28.6 months for FAC-LMS (P = 0.01). Although the median survival for the FAC-BCG-LMS group has not been reached, it is significantly superior to FAC alone (P = 0.009). These three sequential studies suggest that the addition of nonspecific immunotherapy with LMS or the combination of BCG and LMS prolongs the duration of remission and survival of responding patients treated with chemotherapy. Combination immunotherapy with BCG and LMS did not seem superior to LMS alone in this study.


Levamisole as an immunoadjuvant: phase I study and application in breast cancer. A phase I study of levamisole in patients with cancer was undertaken to determine its therapeutic dose range, side effects, limiting toxicity, and influence on immune function. Simultaneously, a group of patients receiving adjuvant combination chemotherapy for carcinoma of the breast were entered in a randomized study to observe whether levamisole was capable of overcoming the immunodepressive effects of cytotoxic drugs. In the phase I trial the doses used ranged from 65 to 410 mg/m2. The maximum tolerated dose was 358 mg/m2. Side effects were generally low in intensity and related to the gastrointestinal and central nervous systems; the limiting toxic effect was intractable nausea. Significant changes in immune test values were noted in both acute and chronic trials. Alterations in immunoglobulin levels, complemented values, and numbers and types of lymphocytes and mitogenic responses to concanavalin A as well as specific mitogens were observed. In the randomized trial there was no difference between the chemoimmunotherapy and chemotherapy-alone groups.


Effect of thymosin and irradiation on immune modulation in head and neck and esophageal cancer patients. Fifty-five patients with squamous cell carcinoma of the head and neck and esophagus were evaluated prior to irradiation and thymosin fraction 5 therapy. Immunity prior to treatment, as measured by total lymphocyte count, E and EAC rosettes, lymphocyte stimulation with phytohemagglutinin (PHA) and in mixed leukocyte culture (MLC) with allogeneic cells, delayed hypersensitivity skin tests, and quantitative serum immunoglobulins, was comparable and normal in the 40 control patients and in the 15 thymosin-treated patients. After irradiation, significant depression (P less than 0.01) was demonstrated in cellular immunity in both groups of patients with decreased T- and B-cell numbers and depressed phytohemagglutinin and MLC stimulation. Six months after irradiation, our preliminary results suggest that the thymosin-treated patients may be reversing their immunosuppression by a return of MLC function and positivity of delayed hypersensitivity skin tests. The ultimate effect of thymosin on disease control and survival remains uncertain.


Therapeutic activity of maleic anhydride-vinyl ether copolymers against spontaneous, autochthonous murine mammary tumors. We have tested a series of five maleic anhydride-vinyl ether copolymers of varying molecular weight for antitumor activity against spontaneous, autochthonous murine breast tumors (average initial tumor weight, approximately 200 mg). Four of the five compounds demonstrated statistically significant inhibition of tumor growth at nontoxic doses. (The fifth compound demonstrated antitumor activity only at toxic doses). To our knowledge, this is the first report of significant activity in a spontaneous solid tumor system in a truly therapeutic setting (ie, with systemic administration only after the autochthonous tumors are clinically evident) with immunologic treatment alone.


Effect of maleic anhydride-divinyl ether copolymers on experimental M109 metastases and macrophage tumoricidal function. Pyran copolymer (NSC-46015) was compared with five maleic anhydride-divinyl ether copolymers (MVEs) of narrow molecular weight range both for the ability to render macrophages nonspecifically tumoricidal and to retard the development of artificially induced metastases. All MVEs were found effective at activating macrophages in vivo, although the optimal dose for each varied. No correlation was obtained between intrinsic viscosity and degree of activation. Pyran was found to strikingly inhibit M109 pulmonary metastases formation when given over a period of 5 days prior to, or 1 day after, iv tumor inoculation. Histologically, tumor inhibition appeared to result from macrophage accumulations and histiocytic granulomas in the lung. Generally, when MVEs were compared in the artificial metastasis model, polymers with the lower molecular weights were the most active.


Potential mechanism of macrophage-mediated tumor cell cytotoxicity. Activated macrophages have been shown to inhibit the incorporation of 125IUDR into the DNA of Lewis lung (LL) tumor cells and to lead to the cytotoxicity of these target cells. Despite this apparent inhibition of DNA synthesis, the increase in the number of LL cells cultured with activated macrophages paralleled that of LL cells alone for the first 18 hours of culture (approximately one doubling of cells). To resolve this paradox, the DNA content of LL cells, which were cultured with activated macrophages for 16 hours, was analyzed by flow microfluorometry. These experiments indicated that subsequent to co-culture with activated macrophages, LL cells with a 50% reduction in their normal DNA content could be detected. Furthermore, after a 2-hour incubation with colcemid, a greater than 95% reduction in metaphase cells occurred in LL cells cultured with activated macrophages. However, dividing nuclei without chromosome condensation appeared to be a prominent feature of these cultures. This would suggest that the genotypic or phenotypic program for cell division in the tumor cells is such that under these unique circumstances of tumor cell-activated macrophage interaction, cell division can proceed in the absence of DNA synthesis. We suggest that this aberrant division may be related to the lethal event which leads to activated macrophage-mediated tumor cell cytotoxicity.


Cell and tissue distribution of 14C-labeled pyran copolymer. The tissue distribution of pyran (maleic anhydride-divinyl ether) copolymer was studied after a single ip injection of 14C-labeled pyran (25 mg/kg) to mice. The pyran showed a reticuloendothelial distribution with the liver and spleen containing the highest concentrations which persisted for at least 21 days after drug treatment. Blood levels of 14C-pyran reached a peak 2 hours after injection and were cleared within 6 hours. Attempts to measure uptake of 14C-pyran by peritoneal macrophages were unsuccessful due to an inability to recover macrophages between 3 and 24 hours after ip pyran administration. Since activated macrophages appear to be the primary mechanism by which pyran enhances host resistance to microbial infection and neoplasia, the uptake of 14C-pyran by isolated peritoneal macrophages in vitro was studied. Purified macrophages showed a gradually increasing uptake of 14C-pyran, and a large amount of cell-associated radioactivity was bound to trichloroacetic acid-precipitable material. Several polyanions, including unlabeled pyran, dextran sulfate, and poly(I)-poly(C), competed for acid-precipitable receptor molecules. The superior antitumor effects of pyran as compared to other polyanions may result from the continuous presence of the synthetic polymer in the host. Possible mechanisms of immunopotentiation by pyran are discussed.


Preliminary observations on the effect of glucan in combination with radiation and chemotherapy in four murine tumors. The efficacy of glucan in combination with local radiation therapy was measured using three solid murine tumors of differing abilities to induce a host defense. Using the KHT fibrosarcoma which induces no measurable host defense, glucan did not improve tumor-free survival over radiation alone; the combination produced a marginal improvement in tumor-free survival in animals bearing the highly immunogenic EMT-6 tumor. The most marked improvement in tumor-free survival was found with the mildly immunogenic 6C3HED lymphosarcoma. The efficacy of glucan in combination with BCNU chemotherapy was measured using the disseminated AKR transplantable leukemia; the combination yielded a high level of cures compared to no survival for either agent alone. Using the AKR transplantable leukemia in an F1 model, the effect of amphotericin B (AmB) alone or in combination with BCNU was tested. AmB or BCNU alone had little or no curative effect when tested in (AKR X DBA)F1 mice, but 56% of mice were cured when combined therapy was employed. When tested in (AKR X C57BL)F1 or (AKR X A)F1 mice, a small fraction was cured with AmB alone while about 90% were cured with either BCNU alone or the combination.


Effect of glucan and other adjuvants on the clearance of radiolabeled tumor cells from mouse lungs. The iv injection of two forms of glucan (DiLuzio and Yamamura), Corynebacterium parvum, or bacillus Calmette-Guérin (BCG) caused a significant increase in the rate and extent of clearance of subsequently injected radiolabeled T1699 carcinoma cells from the lungs of syngeneic DBA2 mice. Dose-response curves were obtained for each agent, the relative rates of clearance of radiolabeled cells were established, and preliminary sequential comparative studies were undertaken. The optimum dose for C. parvum appeared to be lower than for DiLuzio glucan which appeared to be lower than for Yamamura glucan or BCG. The results indicated that at optimum doses one form of glucan (DiLuzio) was as effective as C. parvum and more effective than BCG or a second form of glucan (Yamamura), with respect to the time for the genesis, the magnitude, and the length of time of the response. Very preliminary experiments with a highly malignant and poorly immunogenic cell line of B16 melanoma in syngeneic C57BL/6J mice demonstrated a correlation between an increased level of clearance of radiolabeled cells and a reduced number of subsequent macroscopic lung tumor nodules.


Cytotoxic activity of interferon-treated macrophages studied by various inhibitors. The ability of interferon-treated macrophages to kill or inhibit the growth of tumor cells is markedly influenced by the local environment. The macrophage cytotoxic effector system is regulated by serum and other environmental factors that suppress tumor killing. Prostaglandins E1 and E2, but not F2alpha reversibly inhibited the tumoricidal state of interferon-treated macrophages. Hydrocortisone was similarly active at suppressing macrophage function. Such nonimmunologically derived factors could prevent the final triggering step for macrophage-mediated tumor killing in the local environment of the tumor and may be important in the pathogenesis of progressive tumor growth.


Role of interferon in augmentation of natural and antibody-dependent cell-mediated cytotoxicity. Natural killer (NK) activity in rats and NK and antibody-dependent cell-mediated cytotoxicity in mice have been shown to be rapidly augmented by in vivo treatment with a variety of viruses and immune adjuvants. Poly I:C and other interferon inducers also caused boosting in NK-cell activity, with peak levels occurring at about the time of peak interferon levels. Crude and purified virus-induced interferon itself had strong boosting effects in mice, both when inoculated in vivo and when incubated briefly with normal spleen cells. The in vitro activation by interferon or by poly I:C was completely abrogated by anti-interferon. Interferon also appears to play a central role in augmentation of human NK-cell activity. Inoculation of normal volunteers with influenza virus or of a patient with poly I:C produced rapid boosting. In vitro incubation of peripheral blood mononuclear cells with various interferon preparations for 1 hour or overnight usually produced an appreciable increase in lytic activity.


Human interferon and its inducers: clinical program overview at Roswell Park Memorial Institute. An overview of the clinical interferon program at Roswell Park Memorial Institute is presented. Purified fibroblast interferon and a novel inducer of human interferon [rIn-r(C12,U)n] are being evaluated for possible antiviral, antiproliferative, and immunomodulatory activities in patients with cancer.


Human fibroblast interferon in human neoplasia: clinical and laboratory study. Human fibroblast interferon (HFIF) produced on a large scale from normal diploid cell strains was highly purified and then evaluated as to its safety clinical investigation. The selective antiproliferative activity of HFIF was observed in vitro against certain human malignant cell lines and in vivo against human bladder tumors grown in nude mice. Direct injections of HFIF into metastatic melanoma lesions of two patients resulted in either the disappearance of malignant cells or the significant reduction in tumor volume.


Phase I-II trials of poly IC stabilized with poly-L-lysine. Poly IC, stabilized with poly-L-lysine and carboxymethyl cellulose (poly ICLC), resists hydrolysis by primate serum (unlike the parent compound), induces high levels of serum interferon, and is effective in acute viral infections of subhuman primates. In a phase I-II clinical trial, poly ICLC was given iv in 15 daily doses of 0.5-27.0 mg/m2 to 19 patients with various solid tumors and to six patients with acute leukemia (1-65 years of age). At least three complete trials were conducted at each of six dose levels. Toxic reactions included fever (in 100% of trials), nausea (44%) hypotension (28%), thrombocytopenia and leukopenia (68%), erythema (12%), and polyarthralgia plus myalgia (16%). Hypotension and arthralgia-myalgia were related to dose level and/or magnitude of interferon induction, but other toxic manifestations were not. Poly ICLC induced significant serum interferon levels in 76% of trials, and the correlation between dose and peak interferon titer was linear. The maximum tolerated dose for all patients at a given drug dose was 12 mg/m2; at this dose, the mean peak interferon titer was 1940 reference units/ml. At a dose of 18 mg/m2, the mean peak interferon titer was 4473 reference units/ml, but severe myalgia and arthralgia were intolerable in at least half of the patients, and most had significant hypotension. At a dose of 27 mg/m2, one patient had acute renal failure. At high doses, iv poly ICLC also induced interferon in the cerebrospinal fluid.


Overview of the activity of a Brucella abortus preparation, Bru-Pel. The properties of a nonviable, aqueous ether-extracted Brucela abortus preparation, Bru-Pel, are described. In addition to inducing a "virus-type" interferon response and protecting mice against challenge with otherwise lethal doses of Semliki Forest virus, Bru-Pel is demonstrated to have potent antitumor properties in mice. These antitumor effects appear to be mediated by an increase in nonspecific resistance similar to that seen with other experimental antitumor agents.