Undispensed prescriptions in a mining general practice. Seven per cent of prescription forms issued in a mining practice were not presented at chemists for dispensing. The people least likely to present their prescription forms were men aged 25-34 years, particularly miners. To get sickness benefit these men have to consult a doctor, but the medical content of the consultation in these circumstances is often perceived by them as irrelevant and the medication rejected. Children and old people nearly always presented their prescriptions. The percentage of undispensed prescription forms for the trainee doctor was higher than for the trainer, but age may have been a factor: older patients tended to consult the established doctor, whom they knew. Drugs affecting the cardiovascular system, moderate or strong analgesics, hypnotics, sedatives, tranquillisers, and antidepressants were rarely rejected, but mild analgesics and drugs prescribed for symptomatic relief, such as those affecting the alimentary system, cough mixtures, and skin preparations, were more often rejected.


Transposition of the great arteries: logical anatomical arterial correction. In a 20-month-old child with classic transposition of the great arteries the pulmonary artery, coronary arteries, and aorta were successfully retransposed at arterial level. It was difficult to reimplant the coronary arteries in the dilated pulmonary artery root without damaging the attachments of the valve cusps, and it may be better to divide the pulmonary artery first to avoid this hazard. Retransposition at arterial level will not be suitable for all patients with transposition of the great arteries, and many questions about long-term outcome remain to be answered.


Anatomical correction of complete transposition of the great arteries and ventricular septal defect in infancy. Two patients, aged 8 weeks and 5 years, with D transposition of great arteries and large ventricular septal defect were treated by transection of both aorta and pulmonary arteries and reattaching them to the appropriate ventricles. This included the origins of the coronary arteries. The ventricular septal defect was closed through a transverse ventriculotomy using a Dacron patch. The younger child was operated on as an emergency because of cyanosis and severe heart failure resistant to intensive medical treatment. The older child had had previous banding of the pulmonary artery at the age of 1 year. In both patients pulmonary artery pressure dropped to below half systemic pressure immediately after the operation. Postoperative progress was satisfactory with relief of cyanosis and heart failure. Early anatomical correction of transposition of the great arteries and ventricular septal defect is feasible and should play an important part in the management of these patients.


Dual-demand pacing for reciprocating atrioventricular tachycardia. By using programmed electrical stimulation of the heart and studying the initiation and termination of reciprocating atrioventricular tachycardia two patients with the Wolff-Parkinson-White syndrome were shown to respond rapidly and consistently to fixed-rate pacing. A demand pacemaker was implanted in each patient, having been modified so as to switch into the fixed-rate mode whenever the tachycardia began, thereby terminating the arrhythmia. This appears to be a promising form of treatment in patients with otherwise intractable paroxysmal tachycardia who have been shown by careful study to respond in this way.


Value of gallium scanning in seminoma of the testis. Whole-body scanning using gallium-67-citrate gave consistently accurate tumour localisation in patients with seminoma of the testis. Thirteen out of 15 scans performed in patients with disseminated seminoma in relapse gave good imaging in all disease areas. Scans in patients with teratoma of the testis were less consistently positive; of nine scans performed in patients with disseminated teratoma seven were entirely negative and two scans lightly imaged large disease masses in two patients. In eight patients with combined (seminoma and teratoma) tumours the scan seemed to reflect the dominant tumour type at the time of scanning. In one of these patients the scans changed from positive to negative, being positive when seminoma was the dominant tumour and negative when a teratoma developed. Gallium-67-citrate scanning is useful in managing seminoma of the testis, both for determining the extent of disease present at initial presentation and for routine follow-up. It may be useful in the differential diagnosis of combined tumours when tumour masses are greater than 2 cm in diameter.


Community care service for diabetics in the Poole area. Because of an increasing work load at the hospital diabetic clinic in Poole general practitioners were asked to help in a community care service for diabetics. In this, general practitioners typically care for non-ketotic patients with maturity onset diabetes who are well controlled on diet or diet and oral hypoglycaemic agents, while the clinic concentrates on more difficult cases and screens patients for long-term complications. The hospital laboratory's work load has been eased because patients may attend for two-hour interval blood sugar estimations when they like; most patients appreciate this flexibility and prefer being looked after by their own doctor. The service has not increased the general practitioners' work load unduly, but it has heightened their awareness of diabetes in the community, thus allowing the diabetic consultant to deal with the problems for which he has been trained. The service has thus provided improved care for all diabetics in the Poole area.


Unexplained hepatitis following halothane. Full clinical and laboratory details of 203 patients with postoperative jaundice were submitted to a panel of hepatologists. All patients whose jaundice may have had an identifiable cause were excluded, which left 76 patients with unexplained hepatitis following halothane anaesthesia (UHFH). Hepatitis in 95% of these cases followed multiple exposure to halothane, with repeated exposure within four weeks in 55% of cases. Twenty-nine patients were obese, 52 were aged 41-70, and 53 were women. Thirteen patients died in acute hepatic failure. Rapid onset of jaundice after anaesthesia, male sex, and obesity in either sex were poor prognostic signs. Of the clinical stigmata of hypersensitivity, only eosinophilia was impressive. The UHFH group had a much greater incidence of liver kidney microsomal (LKM) and thyroid antibodies and autoimmune complement fixation than those patients whose jaundice related to identifiable factors. Thirteen of the 19 patients with LKM antibodies also had thyroid antibodies. In six patients retested two to three years later LKM antibodies had disappeared, although thyroid antibodies persisted. Rapidly repeated exposure to halothane may cause hepatitis, but such a complication is probably rare. Possibly obese women with a tendency to organ-specific autoimmunity may be more at risk. Nevertheless, the comparative risks of rapidly repeated halothane or non-halothane anaesthesia cannot be determined from the present data. If alternative satisfactory agents are available halothane should be avoided in patients with unexplained hepatitis after previous exposure, although in three to five patients with UHFH who were re-exposed to halothane jaundice did not recur.


Folate-responsive neuropathy: report of 10 cases. Ten patients with severe neurological disease that was clinically indistinguishable from subacute combined degeneration of the spinal cord were found to have normal serum vitamin B12 levels. All were folate deficient. Specific folate treatment led to significant reversal of the neuropathy. These findings indicate the need to review orthodox concepts of the role of folic acid in maintaining the integrity of the nervous system.


Deep venous thrombosis of the legs after strokes. Part I--incidence and predisposing factors. Forty out of 76 patients (53%) who had suffered a cerebrovascular accident developed deep venous thrombosis of the paralysed leg, as detected with the 125I-fibrinogen technique. A further five also had thrombosis in the non-paralysed leg. A study of many predisposing risk factors provided no help either in elucidating the cause of venous thromboembolism or in identifying patients at risk of DVT as a complication of cerebrovascular accidents.


Deep venous thrombosis of the legs after strokes: Part 2-Natural history. Seven out of 76 patients who had sustained a cerebrovascular accident suffered a pulmonary embolism as diagnosed at necropsy or by unequivocal antemortem criteria. A further five patients had probable embolisation diagnosed only by clinical and chest x-ray criteria. Eleven of these 12 patients had DVT as diagnosed by the 125I-fibrinogen technique. Though 125I-fibrinogen technique has its limitations, thrombosis seemed to be able to develop at several independent sites in the venous system of the leg.


Psychiatric morbidity and the menopause: clinical features. A sample of 114 women from the general population aged 40-55 years were identified as possible psychiatric cases and subjected to a standardised psychiatric interview. Mean ratings for reported symptoms and observed abnormalities were assessed in relation to menopausal status. There was no evidence of any specific combination of symptoms and signs associated with the cessation of menstrual periods, though after the menopause insomnia and hypochondriacal preoccupations were more common. In comparison with matched normal controls have been previous psychiatric illness, and contact with general practitioners was more frequent. Many women developing psychiatric symptoms at the time of the menopause appear to belong to a vulnerable population who are likely to develop symptoms in relation to stress.


Effects of methyldopa on prolactin and growth hormone. The effects of administration of methyldopa on serum prolactin and growth hormone (GH) concentrations in hypertensive patients were studied. Single doses of methyldopa (750 or 1000 mg) significantly increased serum prolactin levels, peak concentrations occurring four to six hours after drug administrations. Long-term methyldopa treatment was associated with threefold to fourfold increases in basal prolactin levels compared with those in normal subjects. In patients treated with methyldopa for two to three weeks the GH response to insulin hypoglycaemia was significantly greater than in normal subjects and untreated hypertensive patients. In contrast, patients treated for prolonged periods (mean 13-4 months) had a GH reponse indistinguishable from normal.


Communications to an antenatal clinic.

Communications to an antenatal clinic. When referring pregnant women to an antenatal clinic many general practitioners and midwives presented details of their care in an incomplete and inconsistent manner. As a result, hospital staff often had insufficient basic data from which to work. We recommend that those providing care decide what information they need and make the rationale of collecting this information the subject of a well-evaluated local education programme.


Twenty-four hour monitoring of heart rate and activity in patients with diabetes mellitus: a comparison with clinic investigations. Heart rate and activity were recorded continuously in 11 selected diabetics during a normal day, and the observations were compared with results obtained in the same patients in the diabetic outpatient clinic 10 months earlier. Both sets of findings agreed well in heart rate variability and postural tachycardia. In patients with well-controlled diabetes simple tests of reflex cardiovascular control produce results that may be useful in following the course of diabetic autonomic neuropathy.


Beta-cell function improved by supplementing basal insulin secretion in mild diabetes. Insulin supplements, predominantly as a constant basal fish insulin infusion, were given to patients with mild diabetes to reduce the overnight fasting glucose level to normal. The basal plasma human insulin levels were reduced to subnormal levels by the infusion, and the insulin response to intravenous glucose was enhanced. The beta-cell in diabetes seems to be in a vicious circle in which an impaired insulin response to glucose produces hyperglycaemia, which stresses beta-cell function, making it more inefficient. A constant basal insulin supplement to induce basal normoglycaemia may benefit beta-cell function in diabetes.


Effect of triamterene on leucocyte sodium and potassium levels in heart disease. Sodium and potassium levels in plasma and leucocytes and the sodium efflux rate constants of leucocytes were measured in patients with congenital heart disease not on treatment, patients with valvular heart disease being treated with digoxin and conventional diuretics, and patients with valvular heart disease receiving digoxin and either conventional diuretics or triamterene or both. The group being treated with digoxin and conventional diuretics showed low cellular potassium levels, low sodium efflux rate constants, and a rise in cellular sodium levels. Patients given triamterene showed a rise in potassium levels in plasma and cells and in the sodium efflux rate constant.


Why don't we prevent childhood accidents? Accidental injury is the most important epidemic in the Western world today, and is especially important as a cause of death and disability in childhood. Many environmental factors are important causes of accidents, but there is no organisation within which doctors, official bodies, industry, and voluntary bodies can pool their experience and co-ordinate their efforts to reduce these environmental risks. A joint committee on childhood accident prevention should be formed in this country similar to that which exists in Sweden.


Effects of legal termination on subsequent pregnancy. Two hundred and eleven patients who had undergone vaginal termination and were pregnant again were investigated; 43-2% had become pregnant within one year of termination. The overall fetal loss in the 211 patients was 17-5% compared with 7-5% in a group matched for parity but consisting of patients who were pregnant after a spontaneous abortion. Altogether 4-3% of pregnancies after legal abortion ended as first trimester abortions, 8-5% as second trimester abortions, and 13-7% in premature delivery. Among 11 women whose cervices had been lacerated at the time of legal termination the fetal loss in subsequent pregnancy was 45-5%, and only one pregnancy went beyond 36 weeks. Routine Shirodkar suture may be beneficial when the cervix is known to have been damaged at legal abortion. Several patients had asked that their general practitioner should not be told of their termination, and such patients may not admit their termination during a subsequent pregnancy, which could thus be jeopardised. No evidence was found to suggest that infants of patients with a history of legal termination are small for dates.


Fetal activity and fetal wellbeing: an evaluation. The clinical value of the 12-hour daily fetal movement count (DFMC) as a test of antepartum fetal wellbeing was assessed. The lowest 2-5% of 1654 DFMCs recorded by 61 women who subsequently delivered healthy infants fell below 10 movements per 12 hours. This level was taken as the lower limit of normal for clinical purposes. A normal DFMC in a population at risk was associated with a satisfactory fetal outcome. A low DFMC was associated with a high incidence of fetal asphyxia, and when fetal death occurred fetal movements rapidly diminished and stopped 12 to 48 hours before death. The DFMC is a generally applicable method of monitoring fetal welfare during pregnancy which provides an inexpensive adjunct or even an alternative to the more expensive placental function tests in current use.


Standardised approach to gluten challenge in diagnosing childhood coeliac disease. Thirty-five children, in whom coeliac disease had been diagnosed on inadequate grounds and who had been on a gluten-free diet for one to 10 years, were challenged with gluten in accordance with a standardised procedure. All children were admitted to hospital for 48 hours for general assessment, two one-hour blood xylose tests, and the introduction of gluten. Thirty children underwent a pre-challenge peroral jejunal mucosal biopsy; the specimens were either normal or showed slight non-specific abnormalities. Gluten powder 20 g/day was given in addition to an otherwise gluten-free diet. The children were reassessed as outpatients every two weeks, when a one-hour blood xylose test was performed. Repeat biopsy was performed when xylose absorption fell or after three months. Seventeen children had abnormal post-challenge biopsy appearances compatible with coeliac disease in relapse; 14 of these children completed their challenge within eight weeks. Seventeen children had completely normal biopsy appearances at the end of three months and were returned to a normal diet. One to two years later eight underwent repeat biopsies, which showed nothing abnormal. In only one child, the oldest in the series, were the histological findings equivocal. In the 17 children in whom coeliac disease was confirmed the duration of gluten challenge was not related to age, duration of gluten-free diet, histological findings on the pre-challenge biopsy, or HLA status.


Can geriatrics survive?

Can geriatrics survive? Geriatrics has consistently failed to attract enough staff, and hence geriatric units often cannot provide a full service for the elderly. Meanwhile beds in general medical units may be blocked by elderly patients. This division between geriatric and medical units is arbitrary and wasteful. There are no clinical processes or techniques unique to geriatrics, which is probably why the specialty is unattractive; few doctors want to confine their attention only to the elderly. Geriatrics as a separate specialty should therefore be largely abandoned and the care of the elderly reintegrated into general medicine.


Gonorrhoea and foreign immigrants at Rotterdam University Hospital. It is estimated, contrary to public opinion and a quick glance at the waiting room of the sexually transmitted diseases (STD) clinic in Rotterdam, that the incidence of gonorrhoea among foreign immigrants is only a little greater than among the Dutch population. The analysis corroborates the common sense idea that the higher incidence of gonorrhoea among foreign immigrants stems largely from their difficulty in finding sexual partners. To solve this problem is a matter of national policy. It is within the scope of the STD clinics and related organizations to start small-scale experiments to improve the difficult medical situation of the foreign immigrants by lowering the linguistic and cultural barriers between the medical staff and their foreign patients. Research on the sexual habits of the patients is of limited value, and a redirection of our efforts seems necessary to tackle this problem.


Climbing fiber responses of cerebellar Purkinje cells to passive movement of the cat forepaw. The activity of cerebellar Purkinje cells during controlled and passive movement of the forepaw was studied in the cat. Burst responses characteristic of activation by climbing fibers were observed in Purkinje cells in lobules Vb and Vc of the cerebellar vermis and paravermis. The climbing fiber responses followed the onset of a movement with a latency ranging from 20 to 60 msec depending upon movement type and amplitude. Responsive Purkinje cells were localized in a well defined parasagittal strip very near the paravermal vein in lobules Vb and Vc. Cells within the responsive strip responded with identical response probabilities and latencies for any particular type of movement presentation. Responses were independent of starting paw position and direction of movement. Climbing fiber responses could be evoked by extremely small movements with most cells responding to displacements of 50 mum. The latencies and probabilities for climbing fiber responses were inversely related to movement amplitude with latencies as long as 80 msec for very small displacements.


Nicotinic effect of acetylcholine on the release of newly synthesized (3H)dopamine in rat striatal slices and cat caudate nucleus. The effect of acetylcholine (ACh), carbachol and nicotinic blocking agents on the release of newly synthesized [3H]dopamine ([3H]DA) was studied in vitro on rat striatal slices and in vivo on the cat caudate nucleus. In the latter case, the animals were anaesthetized with halothane; in some experimetns an 'encéphale isolé' preparation was used to eliminate anaesthesia. Rat striatal slices placed in a superfusion chamber were continuously superfused with L-[3,5-3H]tyrosine. A cup placed on the ventricular surface of the cat caudate nucleus similarly allowed a continuous superfusion of the structure with the 3H amino acid. In both cases the quantities of [3H]DA contained in serial superfusate fractions were estimated; the drugs were always added in superfusing medium. In vitro ACh (10(-5) M) and carbachol (10(-5) M) enhanced the release of [3H]DA (90%). Similar results were obtained in vivo in anaesthetized cats. The effect of ACh (10(-5) M) was more pronounced (125%) in presence of eserine (10(-4) M) than with ACh alone (65%). ACh was also effective in unanaesthetized cats. The ACh effect on [3H]DA release was reproducible within the same experiment both in vitro and in vivo. This allowed to test the effect of anticholinergic agents on the ACh induced release of [3H]DA. In vivo hexamethonium (10(-4) M, 10(-5) M) partially blocked the release of [3H]DA induced by ACh (10(-5) M) alone; the effect was not seen when ACh was added in the presence of eserine (10(-4) M). Both in vivo and in vitro the prior introduction of mecamylamine into the superfusing medium antagonized the stimulating effect of ACh (10(-5) M) on [3H]DA release. The effects of this nicotinic blocking agent were seen with various concentrations (10(-6); (10(-5) 10(-4) in the in vitro experiments.


Dopaminergic innervation of the rat prefrontal cortex: a fluorescence histochemical study. After the destruction of the noradrenergic ascending pathways, the localization of frontal cortical fields receiving fibers from the dopaminergic mesocortical system has been studied in rats, using a glyoxylic-paraformaldehyde method. The dopaminergic innervation was distributed in two main areas. The area of highest density was a medial field which spread in the medial cortex anterior and dorsal to the genu of the corpus callosum. It did not reach the shoulder region except in the foremost part of the frontal lobe where dopaminergic fibers were scattered in the whole cortex, including the molecular layer. A deep sulcal field was situated between the dorsal bank of the rhinal sulcus and the lateral cortex above it. In addition, a moderately dense band of dopaminergic fibers was observed between the corpus callosum and the anterior commissura, beside the accumbens nucleus. Similar data were obtained with dopamine uptake experiments on reserpine-treated but otherwise normal animals. The frontal areas receiving dopaminergic innervation coincide strikingly with the 'prefrontal cortex' as defined by neuroanatomical studies, which is assumed to be more or less equivalent to the prefrontal cortex of primates and derives direct projections from the amygdala. The functional implications of these findings are discussed.


Habituation of exteroceptive suppression and of exteroceptive reflexes in man as influenced by voluntary contraction. Habituation kinetics were found to vary considerably in different polysynaptic pathways of the brain stem in man. The exteroceptive reflex of the digastric muscle habituates markedly, even for stimuli repeated at 1-min intervals. The second component of the blink reflex habituates for intervals below about 8 sec, but this can largely be prevented by steady voluntary contraction of the muscle. On the other hand the exteroceptive suppressions of the masseter muscle only disclose slight habitaution of their late component ES2 and no habituation of the early component ES1. This does not appear to be affected by voluntary activation since the suppressive effects present the same pattern in relaxed masseter muscles tested by proprioceptive reflex activation. Exteroceptive reflexes and exteroceptive suppressions of similar latencies disclose genuine and important differences in their habituation kinetics.


Brain stem and cerebellar lesions in female rats. I. Tests of posture and movement. Lesions were placed in a number of brain stem and cerebellar structures to determine their importance to the control of posture and movement in rats. Lesions of the lateral vestibular nucleus, superior vestibular nucleus, or inferior olive produced severe disturbances of posture and movement. Rats with such lesions had pronounced tremor, abnormal head posture and asymmetrical trunk posture, and failed in a balance beam test. Lesions of the medial medullary reticular formation and medial vestibular nucleus had less overall effect on posture and movement, while animals with lesions of the deep cerebellar nuclei were relatively unimpaired. All lesion groups showed some recovery of function during the course of postoperative testing.


Brain stem and cerebellar lesions in female rats. II. Lordosis reflex. The effects of brain stem and cerebellar lesions upon the lordosis reflex were studied. Lesions of the lateral vestibular nucleus (LVN) and nucleus reticularis gigantocellularis (NGc) produced significant deficits in lordosis. Lesions of the deep cerebellar nuclei, inferior olive, medial vestibular nucleus, and superior vestibular nucleus did not produce significant effects. Results from animals with lesions of both lateral vestibular nucleus (LVN) and nucleus gigantocellularis (NGc) suggested that these structures act independently in their control of lordosis. Histological analysis showed that giant cell loss in NGc and Deiters' cell loss in LVN were both significantly correlated with decrements in lordosis. These results indicate that lateral vestibulospinal and reticulospinal pathways participate in the descending control of segmental mechanisms responsible for the lordosis reflex.


Striate cortex ablation and spatial vision. Normal and visually decorticate rats were trained to orient to a luminous target. All subjects reached the same criterion of orientation accuracy, but operated subjects required more training and exhibited greater response latency throughout testing. To determine if visually decorticate subjects located the target according to the retinal position of its image, or only by sampling luminous flux density at different points in space, field-displacing wedge prisms were placed before the subjects' eyes to laterally displace the retinal locus of the target image without affecting luminous flux information. Prism imposition shifted orientation toward the virtual target position for all subjects. In a second study, translucent plastic was place over the subjects' eyes to disrupt the retinal topography and location of the target image while again leaving luminoux flux unaffected as a potential cue. All animals showed a dramatic and complete disruption of target localization once the diffusing material was applied. The data were considered as support for the hypothesis that 'spatial' vision survives visual decortication.


A qualitative electron microscopic investigation of the anomalous corticofugal projections following neonatal lesions in the albino rats. The present study confirms the suggestive evidence brought forth by the finding of Leong and Lund. Following neonatal lesion of the sensorimotor and adjacent cortex of the albino rats, the remaining intact contralateral cortex projects bilaterally to the superior colliculus, pons and spinal cord, all of which normally receive a unilateral corticofugal projection. Like the normal projections to the 3 areas studied, the anomalous projections form Gray's type I axo-dendritic relations. The terminals of the anomalous projections seem to form a more complex configuration with one axon terminal synapsing on more dendrites than is usual.


The distribution of free amino acids in subdivision of rat and frog retinae obtained by a new technique. A technique is described for splitting intact rat and frog retinae into laminae. These laminae have been examined histologically and the distribution of free amino acids in the laminae has been determined by the dansylation technique. In both rat and frog retinae, taurine was selectively concentrated in the photoreceptor cells and their processes. In the rat, glycine, glutamine and GABA were especially abundant in a lamina which contained the ganglion, inner plexiform and inner neuronal layers. High concentrations of GABA were also present in the outer plexiform layer. These results are discussed in terms of a possible transmitter function for these amino acids in the retina.


Temporal sequence of frequency specific and nonspecific effects of flickering lights upon the occipital electrical activity in man. Human subjects were exposed 10 times to light flicker for a period of 10 sec, being altered with equal non-flicker periods. Four flicker frequencies were studied: the resting alpha frequency, twice the frequency, and 14 and 6 c/sec, which are both unrelated to alpha rhythms. The effect of flicker stimulation upon the occipital EEG was quantified in terms of integrated values per band over an epoch of 10 sec using a Walter-type frequency analyzer.


The afferent input and functional organization of the hypothalamus in reactions regulating pituitary-adreno-cortical activity. In chronic experiments on rabbits, stimulation of the dorsal tegmentum of the midbrain caused an increase in serum corticosteroid level. In acute experiments, stimulation of the same region resulted in changes in the activity of hypothalamic neurons. Neurons in the anterior (AHA), medial (MHA), including the medial dorsal (MHAd) and medial ventral (MHAv) areas, and lateral (LHA) areas of the hypothalamus were studied. When the midbrain was stimulated, more neurons in LHA, MHAd and MHAv were excited than inhibited; in AHA, more neurons were inhibited. These changes suggest that the activation of the pituitary-adrenocortical system caused by stimulation of the dorsal tegmentum is associated with changes in the influx of afferent impulses to the hypothalamus, which are responsible for AHA inhibition and MHA and LHA excitation.


Influences of hypothalamic stimulation upon septal and hippocampal electrical activity in the cat. Spontaneous patterns of hippocampal EEG and septal cell activity were studied in immobilized cats, and the influences of high frequency stimulation of medial hypothalamus (MH) and lateral hypothalamus (LH) were determined. Septal cells were divided into 3 classes on the basis of their discharge patterns: (1), rhythmic bursting (2), non-rhythmic bursting and (3), non-bursting, and the relationship of these discharge patterns to hippocampal theta rhythm was analyzed. Rhythmic bursting cells displayed close frequency and phase relations to hippocampal theta rhythm and were located chiefly in the diagonal band of Broca. Cells of the other two categories were found both within and outside of the diagonal band region.


Retinofugal projections in the opossum, An anterograde degeneration and radioautographic study. A study of anterograde degeneration and anterograde transport was undertaken in the opossum primary optic system in order to clarify several points regarding fiber organization and patterns of terminal fields. Through the radioautographic technique of axon tracing, it was demonstrated that the accessory optic system follows the generalized scheme of Hayhow, consisting of two fascicles the three terminal nuclei.


Undernutrition in the developing rat: effect upon myelination. Long-Evans rats were undernourished from birth by removing the mother from the sucling rats for part of each day; the undernutrition was continued after weaning till 60 days of age by restricting the daily food intake. Brain development was monitored by histologic and selected biochemical analyses coordinated with an ultrastructural morphometric analysis of the pyramidal tract of 30-day-old rats. Brain and body growth were already reduced after 10 days of undernutrition. At 20 days of age, the peak of myelination in the controls, the body and brain weights of the undernourished rats, compared to the controls, were reduced by 25 and 60%, respectively, and by the end of the study (60 days), the brain and body weights were reduced by 30 and 70%, respectively. Morphometric analysis indicated that the proportion of myelinated axons was significantly reduced in starved (34%) relative to control (43%) animals. Fiber analysis revealed that not only were the myelin sheaths thinner in the undernourished rats, but that the sheath was disproportionately reduced relative to the axon diameter. Chemical analysis on a whole brain basis demonstrated a greater than 60% deficit in the relatively myelin-specific galactolipids. (Whole brain analysis included regions more severely affected than the morphometrically analyzed pyramidal tract.) We also obtained evidence for both a delay in the initiation and a general retardation of myelinogenesis. The promyelinating fibers (axons with one or two non-compacted myelin lamellae) still constituted 4.5% of the myelinated fibers in the undernourished animals at 30 days but had declined to 0.3% in the controls. Analysis of the fabtty acids of cerebroside and sulfatide (lipids enriched in myelin) demonstrated in the undernourished group a pattern characteristic of a younger animal. Thus, the effect of undernutrition on the developing rat appears to be one of inhibited and somewhat retarded myelination. These effects were most likely due to a reduction in the number of myelinating glia formed and the restricted capacity of those which form to generate myeline lamellae.


Myelin synthesis during postnatal nutritional deprivation and subsequent rehabilitation. Newborn Long-Evans rats were undernourished by maternal deprivation so that by 20 days of age their body and brain weights were about 45 and 80%, respectively, of the values obtained for control (well-nourished) values. Proteins from myelin of undernourished and control rats were separated by polyacrylamide gel electrophoresis in buffers containing sodium dodecyl sulfate. At 15 and 20 days of age the proportion of basic and proteolipid protein was reduced in the starved animals relative to controls, indicative of a delay in maturation. However, by 30 days of age the composition of myelin from starved and control animals appeared similar. At all ages the yield of myelin from brains of starved rats was less than 25% of that obtained from control animals. A series of isotope labeling experiments, using a double label design, was carried out to compare relative rates of incorporation of radioactive amino acids into individual proteins of various brain subcellular fractions. In 20-day-old rats the incorporation of [3H] OR [14C] leucine or glycine into myelin proteins, relative to incorporation into proteins of other subcellular fractions, is preferentially depressed (about 60%) in starved animals. Synthesis of all the myelin proteins was depressed, supporting the hypothesis that the high molecular weight proteins isolated with myelin are true myelin constituents. Similar experiments were conducted using [3H]-and [14C] acetate, choline, or glycerol as precursors of lipids. Incorporation of isotope into lipids of myelin, relative to lipids of other subcellular fractions, was also depressed by about 60% in starved animals. In several experiments we studied synthesis during rehabilitation (ad libitum feeding) following 20 days of postnatal starvation. After 6 days of rehabilitation, incorporation of radioactive precursors into myelin, relative to other subcellular fractions, was still depressed. This result was true for both proteins and lipids, and was interpreted as evicence against the initiation of a process leading to a net recovery of myelin (i.e., an irreversible deficit of myelin synthesis is induced by this regime of nutritional deprivation).


Some properties of the transmitter release mechanism at the rat ganglionic synapse during potassium stimulation. In the present investigation a study has been made using intracellular recordings in the rat superior cervical ganglion of the mode of transmitter release induced by raised external potassium ion concentration (40 mM), after acetylcholine synthesis has been blocked by hemicholinium-3. It is shown that the progressive decline in the rate of acetylcholine output from the ganglion is related to a decrease in the number of quanta being released. Furthermore, under these conditions there is no evidence for a reduction in the size of the transmitter quantum. The statistical foundations of the quantal release process at the rat ganglionic synapse have been investigated by comparing the distribution of the number of miniature EPSPs during successive constant time intervals in the tracings with the corresponding Poisson and binomial predictions. Analyses have shown that the probability for a quantum to be released is so small as to produce a binomial distribution of responses indistinquishable fro- the corresponding Poisson distribution, both at the beginning of the potassium-induced quantum discharge and when transmitter release level is low after exhaustion of acetylcholine tissue content.


Topographical distribution of dopaminergic innervation and of dopaminergic receptors in the rat striatum. I. Mictoestimation of [3H] dopamine uptake and dopamine content in microdiscs. Topographical variations in the uptake of [3H] dopamine (DA) and in the endogenous content of DA were estimated in the striatum of the rat. For this purpose, microdiscs were punched out in serial 500 mum sections. [3H] DA uptake was measured in 0.25 M sucrose homogenates prepared from microdiscs punched out from frozen slices (--7C). This uptake was similar to that observed in fresh tissues. It was unaffected by desmethylimipramine (5 X 10(-7) M), inhibited by benztropine (10(-6) M) and no longer detectable after 6-hydroxydopamine-induced degeneration of the nigrostriatal dopaminergic pathway. Both [3H] DA uptake and DA content decreased regularly from the rostral to the caudal part of the structure. In contrast, no important differences could be found in the dorso-ventral plane. These results suggest that the extent of dopaminergic innervation is heterogenous within the structure.


Topographical distribution of dopaminergic innervation and of dopaminergic receptors in the rat striatum. II. Distribution and characteristics of dopamine adenylate cyclase--interaction of d-LSD with dopaminergic receptors. The characteristics of dopamine adenylate cyclase in the rat striatum were first studied on homogenates of fresh tissues. In the assay conditions used, dopamine (10(-4) M) stimulated the enzyme activity by 250%. This effect was completely blocked by fluphenazine (10(-5) M; Ki=9X10(-9) M) and by phentolamine (10(-5) M; Ki=3 X 10(-7) M). D-LSD stimulated the adenylate cyclase activity (Km=1.4 X 10(-7) M) by interacting with dopamine receptors; indeed the dopamine effect on the enzyme activity was competitively reduced in presence of D-LSD. L-Isoproterenol (Km=10(-6) M) activated an adenylate cyclase through a receptor distinct form the dopaminergic receptor; this stimulation was not affected by fluphenazine or phentolamine but suppressed by DL-propranolol (10(-4) M). The topographical distribution of the dopamine, D-LSD and L-isoproterenol adenylate cyclase activities were examined in homogenates prepared from discs punched out on serial frozed (--7C) slices of the striatum. Under this condition, tge dioanube naxunak stunykatuib was if 150%. A 4.8-fold progressive decrease in the amount of cyclic AMP produced in presence of dopamine (10(-4) M) was observed in the rostrocaudal plane of the structure; the decline of the basal activity was 3.6-fold. The topographical curves of maximal activation of adenylate cyclase by dopamine and D-LSD were superimposable confirming that D-LSD acts on dopaminergic receptors. This topographical distribution of dopamine sensitive adenylate cyclase is comparable on one hand to that of endogenous dopamine and on the other hand to that of the dopamine high affinity uptake activity measured in simultaneous experiments. In contrast to that observed with dopamine or D-LSD, the topographical distribution of the adenylate cyclase sensitive to L-isoproterenol was homogenous within the striatum.


Metal chelates of L-DOPA for improved replenishment of dopaminergic pools. An exploratory study consisting of physiochemical and animal experiments was undertaken with the objective of developing one or more metal-L-DOPA chelate systems for an improved transport of L-DOPA into the brain. This approach is based on a theoretical speculation that the pyridoxal-dependent decarboxylation of L-DOPA in the precerebral areas might be obviated by an appropriate metal chelation of the aminocarboxylate end of the L-DOPA molecule. Equilibium studies on the interactions of L-DOPE with Cu2+,Zn2+, Co2+, Mg2+ and Fe2+ ions and their ATP chelates were carried out in order to examine the conditions for the selective binding of the terminal amine group. Metal chelate systems for in vivo transport experiments were selected viz., Cu2+ or Zn2+-L-DOPA (1:2) and Cu2+ or Zn2+-ATP-L-DOPA (1:1:1) which contained the amine-bound metal ion in a completely coordinated form. Results of in vivo studies involving the intraperitoneal administration of 14C-and 3H-LABELED L-DOPA compounds have shown a 100-150% increase in the transport of L-DOPA into the brain by using the Cu2+ and Zn2+ chelates over that effected by using the unchelated L-DOPA. A chromatographic analysis of the brain homogenates showed that only 6% of the overall radioactivity of the brain could be attributed to 3-methoxytyrosine, and the remaining activity was due to DOPA, dopamine and norepinephrine. The transport effectiveness was also compared with that obtained by using the combination drug, RO4-4602 + L-DOPA.


Basal forebrain and hypothalamic influences upon brain stem neurons. Intracellular recordings were performed from bulbar and caudopontine (BcP), rostropontine (rP) and mesencephalic (Mes) neurons in acute encéphale isolé preparations during low- and high-frequency basal forebrain (BF) and hypothalamic (Hyp) stimulation. Low-frequency BF stimulation induced long-latency (7-8 msec) EPSPs on 31% of BcP neurons. High-frequency stimulation of the same regions induced excitation in 58% of them. No inhibition was observed. The descending influences on BcP neurons disappeared after bilateral acute sensorimotor decortication. Low- and high-frequency BF stimulation influenced a very low percentage of rP and Mes neurons. The only effect was a long-latency slow rising EPSP with or without spikes. No inhibition was found in these neurons. Low- and high-frequency Hyp stimulation produced short-latency EPSPs with or without spikes on a large percentage of BcP, rP and Mes neurons. The effect was more constant on rostral (rP) than on caudal (BcP) neurons and more evident when high-frequency stimulation was applied. At high frequency the activation of brain stem neurons paralleled an EEG desynchronization. IPSPs were seen in a low percentage of Mes and rP neurons. Mes neurons could be also activated antidromically. The results suggest that BF regions do not antagonize directly the mesencephalic activating system and may induce electrocortical synchronization through the activation of caudal brain stem neurons. This circuit involves sensorimotor cortical neurons. The facilitatory effect of Hyp stimulation on rostral brain stem neurons indicates that the EEG desynchronization which follows high-frequency Hyp activation may be mediated by the ascending reticular system. Other mechanisms mainly at the diencephalo-telecephalic level may however be involved in this effect.


Effects of alpha-toxins from Bungarus multicinctus and Bungarus caeruleus on cholinergic responses in Aplysia neurons. The effects of alpha-bungarotoxin from Bungarus multicinctus and Bungarus caeruleus were studied on three types of cholinergic response in Aplysia central neurones. These responses are the result of three distinct changes in ionic permeability: selective increases in permeabiliyt to Na, Cl and K, respectively. It was shown that 10(-5) M alpha-bungarotoxin from B. multicinctus completely blocks the response resulting from an increase in Cl permeability, while having no effect on either of the other two responses types (even when the toxin concentration was increased to 5 X 10(-5) M). The block of the Cl-dependent response by alpha-bungarotoxin is reversible. None of the three response types were affected by similar concentrations of alpha-toxin from B. caeruleus. Higher concentrations were not systematically tested. These results contradict reports of other authors on the action of alpha-bungarotoxin on molluscan acetylcholine (ACh) responses. Possible reasons for the discrepancies between our findings and those published by other authors are discussed.


Selective neuronal accumulation of omega-amino acids in the rabbit retina. The uptakes of radioactive glycine, beta-alanine, GABA, delta-aminovaleric acid and epsilon-aminocaproic acid were studied in the rabbit retina in vivo and in vitro. Autoradiography demonstrated that the amino acids were mainly taken up and retained by certain cells of amacrine type. In the neurons, the omega-amino acids seem to be stored in a protected fashion. The uptake of delta-aminovaleric acid and epsilon-aminocaproic acid was temperature dependent, saturable, and of high-affinity type (with Km for delta-aminovaleric acid = 3.02 X 10(-5) M and for epsilon-aminocaproic acid 9.38 X 10(-5) M). The uptake of both delta-aminovaleric acid and epsilon-aminocaproic acid was competitively inhibited by GABA, but not by glycine or a number of other amino acids regarded as possible neurotransmitters. The uptakes of delta-aminolevulinic acid, taurine and alpha-amino-isobutyric acid were studied for comparison, but no selective neuronal uptake was observed. It is concluded that there are certain amacrine cells which selectively accumulate delta-amino acids such as beta-alanine, GABA, delta-aminovaleric acid and epsilon-aminocaproic acid. It seems possible that the mechanism accumulating GABA may also work with the other omega-amino acids. This opens the possibility that the omega-amino acids may serve as 'false neurotransmitters'.


Decarboxylation of newly formed DOPA by caudate nucleus synaptosomal particles. The present study compares dopamine formation from two different sources of DOPA: preformed and added to the medium and that newly formed by the hydroxylations of phenylalanine or tyrosine. Synaptosomal preparations from rat brain caudate nucleus were incubated with 3H-labeled DOPA mixed as cosubstrate with either [14C]phenylalanine or [14C]tyrosine. Following the incubation, DOPA and dopamine were separated and their isotope ratios (isotope in phenylalanine (tyrosine) X 100/isotope in DOPA cosubstrate) were determined and compared. The results show that the ratio in dopamine was not only different from that in DOPA but was in fact 8.3-15.0 times higher. Although the absolute values of the isotope ratios were affected by any changes of the cosubstrate concentrations, the ratio in dopamine was found to be higher than in DOPA at all the substrate concentrations tested. When the synaptosomes were separated post-incubation from the medium and analyzed, the ratio in dopamine was also found to be higher than that in DOPA. Also, the large difference between the intrasynaptosomal ratios persisted throughout the incubation period ranging from 10 to 45 min. The results suggest that the DOPA which is newly formed by the hydroxylations, (a) may not be freely exchangeable with added DOPA because of compartmentation and (b) may be preferentially decarboxylated to dopamine.


Some molecular properties of rat-liver lysosomal beta-glucuronidase isoenzymes. The isoenzymes of rat-liver lysosomal beta-glucuronidase (beta-D-glucuronide glucuronosohydrolase (EC 3.2.1.31)) were inactivated at different rates at 0 degrees C in 3M guanidinium chloride solutions adjusted to pH 5.0 In 4 M urea buffered by 0.01 M glycylglycine, pH 7.0 isoenzymes I, III, and V were reversibly inhibited 80%. Sodium dodecyl sulfate (SDS), 0.1% in 0.01 M phosphate buffer, pH 7.0 irreversibly inhibited at 37 degrees C all five isoenzymes. Sedimentation analysis showed that loss of catalytic activity in these denaturing media is accompanied by dissociation into slower sedimenting subunits. SDS gel electrophoresis revealed that the isoenzymes are apparently tetramers made up of different proportions of subunits alpha, beta, and gamma having apparent molecular weights of 62,900, 60,200, and 58,700, respectively. The three subunits appear to be glycoproteins.


Studies of the metabolism of asialotransferrins: nonspecific changes in the metabolic behaviour of human asialotransferrin in avians. In a previous study (Regoeczi, E. & Hatton, M.W.C. (1976) Can J. Physiol. Pharmacol. 54, 27-34) it was shown that the chicken (Gallus domesticus) does not possess a hepatic asialoglycoprotein receptor. In the present study the same conclusion is drawn for the duck (Anas platyrhynchos). For this reason these avian species were used to assess those changes in the distribution and catabolism of human asialotransferrin which takes place in the absence of the asialoglycoprotein receptor of the liver.


Relative importance of alternative pathways of purine nucleotide biosynthesis in Ehrlich ascites tumor cells in vivo. Several alternative pathways of purine nucleotide synthesis coexist in cells and the relative importance of each pathway for maintaining purine nucleotide concentrations in cells have been studied. Specific inhibitors were used to block these synthetic routes in Ehrlich ascites tumor cells in vivo and the effect of inhibiting each pathway was evaluated by measuring intracellular purine nucleotide concentrations by high-speed liquid chromatography. The results of this study indicate that adenosine triphosphate and guanosine triphosphate concentrations of Ehrlich ascites tumor cells are maintained primarily by purine biosynthesis de novo although other pathways do make significant contributions.


Acid phosphatases: androgen dependent markers of rat prostate. Our investigations on acid phosphatase (AP) were aimed at finding a biochemical assay marker for androgen actions in the rat prostrate. We quantitatively examined the effects of l-tartrate or formaldehyde on AP activity in tissue filtrates from nine adult male rat tissues, plasma and hemolysed red blood cells (HRBC). There was significant inhibition of AP activity in all instances with the exception of HRBC with tartrate. The prostate inhibition results were not different from those for seminal vesicles and adrenals but were different from the other tissues studied. Ten days following castration the inhibition by tartrate was less in all tissues studied except plasma and HRBC; the formaldehyde inhibition percentages were not altered.


The future of sensory aids for the blind. Although it is apparent that a successful substitute for vision will not be available for at least two or three decades, the past twenty five years of research is now bearing fruit. Within the next few years several high-technology devices will solve the problems of mobility and reading. Besides this there is a wide range of aids to solve an expanding variety of vocational applications. Great improvements can be expected in the field of closed circuit television and light amplification which will help particularly those with low vision.


The location of conduction abnormalities in human entrapment neuropathies. Direct stimulation of 23 median, 13 ulnar and 2 peroneal nerves at the time of surgical exploration has been used to locate, and characterize the conduction abnormalities in the nerves. The most frequent location of the major conduction abnormalities in the median nerve was in the first 1-2 cm distal to the origin of the carpal tunnel. In the ulnar nerve the important conduction abnormalities were located most frequently in the segments 1 cm proximal and distal to the medial epicondyle. In the peroneal nerve the major conduction abnormalities occurred proximal or distal to the entry point of the common peroneal nerve into the peroneus longus muscle.


Classification of aphasic phenomena.

Classification of aphasic phenomena. A brief but comprehensive survey of classifying aphasia reveals that most investigators describe at least four major groups, conveniently labelled Broca's, Wernicke's, anomic and global. Conduction and transcortical aphasias are less generally described and modality specific syndromes rarely, if ever, exist purely. The controversy between unifiers and splitters continues but objective numerical taxonomy may solve some of the problems of classification.


Integrative versus delay line characteristics of cerebellar cortex. In order to determine which of two general models ("tapped delay line" or "integrator") provides a more accurate desciption of mammalian Purkinje cell (P-cell) activation by natural stimulation, the spatial and temporal characteristics of a population of neurons in cerebellar cortex responsive to small controlled stretches of forelimb muscles were examined in awake, locally anesthetized cats. Stretch of a single wrist muscle excited P-cells over a distance of about 1 mm in the long axis of a folium, a span which is at most half the length of parallel fibers. Both granule cells and molecular layer interneurons were excited over a wider zone than P-cells. Furthermore, P-cells across a response zone all fired on the average at the same time, as determined by computing peristimulus cross-interval histograms from pairs of simultaneously recorded neurons. Consistent delays could only be demonstrated in the minimal response latencies as measured from peristimulus time histograms. These delays, however, were longer than could be ascribed to parallel fiber conduction velocity. No evidence, therefore, was found in cat cerebellum to support the "tapped delay line" model, which postulates the successive activation of P-cells as an excitatory volley travels along a parallel fiber beam. Instead, an integrative mode of operation seems to predominate: a relatively wide substratum of activated granule cells simultaneously activates a narrower focus of P-cells centrally situated with respect to the granule cell population. The role of inhibitory interneurons in promoting the "integrator" model is discussed.


Direct leukocyte migration inhibition in multiple sclerosis--a possible assessment of activity. Twenty-four patients with multiple sclerosis were evaluated and classified according to their clinical state. Specific migration inhibition studies were carried out on blood samples from each using myelin basic protein, an acid soluble protein fraction isolated from normal human CNS white matter, and multiple sclerosis myelin basic protein isolated from patients who had the disease, as the antigenic material. This test system employed selected media. Results were compared with those of normal controls and patients with other neurological disease states. Antigen concentration of 500 mu g/ml in serum free medium combined to produce the greatest inhibiting effect on leukocytes in patients with apparent multiple sclerosis and differentiated these patients from those in the other groups tested. Leukocytes in patients who had probable multiple sclerosis with partial impairment, signifying possible current activity of the disease, were especially inhibited as compared to the leukocytes from other groups tested against myelin antigen. Cerebrospinal fluid from affected patients when used as a media enhanced the test. This study suggests that migration inhibition of peripheral leukocytes using myelin protein may be useful in the diagnosis of patients with multiple sclerosis. There is additional evidence that the degree of leukocyte migration inhibition may reflect activity of the disease with its consequent implications on treatment and prognosis.


Clinical and laboratory findings in primary generalized and multiple-myeloma-related amyloidosis. Thirty-four patients with primary generalized amyloidosis (PGA) and 14 with multiple-myeloma-related amyloidosis (MRA) were studied. The commonest clinical manifestations in PGA were nephrotic syndrome, hepatomegaly and congestive heart failure, and in MRA, low back pain, plasmacytoma and rheumatoid-arthritis-like syndrome. Eight patients with PGA had limited clinical expression of the disease, such as involvement of only kidneys, joints, parotid glands or gastrointestinal tract; in one patient amyloidosis was limited to lymph nodes. Low serum concentrations of total protein and albumin were common. M components were detected in the serum of 91% of patients with PGA and 92% of patients with MRA: 70% of the M components in PGA and 25% of those in MRA had lambda light chains. Bence Jones proteinemia was detected in 56% of the patients with PGA and in 77% of those with MRA. The serum concentration of immunoglobulins was decreased substantially in more than two thirds of the patients with PGA. Proteinuria (greater than 250 mg/24 h) was observed in 78% of patients with PGA and in 93% of patients with MRA. Bence Jones proteinuria was noted in 75 and 77% of patients, respectively. Plasmacytic infiltration of the bone marrow was found in 90% of the patients with PGA. The mean survival time of the patients with PGA was 28 months and of those with MRA, 29 months from the time of diagnosis.


Cryocrystalglobulinemia.

Cryocrystalglobulinemia. A 40-year-old woman had cryocrystalglobulinemia with IgG2(K1). To date, 27 other cases of spontaneous crystallization of a plasma protein have been reported. In all, the protein, a cryoglobulin, has been found to be an IgG molecule. The disease most commonly associated with this phenomenon has been multiple myeloma. None of the patients have had Raynaud's phenomenon, but many have had purpuric skin lesions made worse by exposure to cold. In the two cases of essential cryocrystalglobulinemia, crystals were found in the peripheral blood film. Immunologic, biochemical and ultrastructural studies have so far not demonstrated any property common to all cryocrystalglobulins.


Constipation and catharsis.

Constipation and catharsis. Constipation is endemic in the Western world. Stool consistence and associated symptoms are more important than stool frequency. The patient's attitude towards his bowel habit is also important. Exclusion of "organic" disease, reassurance and discussion of normal variations of bowel habit are necessary first steps in treatment. Adquate bulk in the stool must be ensured, either through diet or bulking agnets, and bowel retraining attempted. Laxatives should be reserved for episodes of constipation following enforced bedrest, or as a preparation for diagnostic procedures. Long-term use should be avoided.


The diabetic day-care unit. II. Comparison of patients and costs of initiating insulin therapy in the unit and a hospital. Fifteen patients began insulin treatment in a diabetic day-care unit and 14 patients began insulin treatment in hospital. The cost per patient of initiating insulin treatment in hospital was nine times the cost in a day-care program. In general, the disease of patients with juvenile-onset diabetes was initially under poorer control in those admitted to hospital than in those in the day-care program. However, the day-care juvenile with disease under poorest control was comparable to the most severely affected patient admitted to hospital. No major differences in the improvements in control achieved by the two programs were found.


Juvenile rheumatoid arthritis: follow-up into adulthood - medical, sexual and social status. Fifty-eight adults were studied an average of 14.5 years after the diagnosis of juvenile rheumatoid arthritis had been made. The pattern of disease had been typical with respect to various categories, complications and course; HLA-B27 antigen was found at follow-up in 25% of those tested. Two thirds now had mild to moderate disease, good sexual adjustment and "normal" educational achievement, employment history and lifestyle. One third had severe disease, often with progressive disability; this did not prevent sexual activity but caused some limitations, and the need for sexual counseling is emphasized. Ongoing activity of the disease process rather than severity of disability seemed the chief factor preventing a normal lifestyle.


Tennis elbow and the cervical spine.

Tennis elbow and the cervical spine. The exact cause of tennis elbow, a common condition, is still obscure. While the condition may well be entirely due to a local disorder at the elbow, the results of a study of 50 patients whose condition was resistant to 4 weeks of treatment directed to the elbow suggest that the underlying condition may have been (at least in these patients) a reflex localization of pain from radiculopathy at the cervical spine. Clinical, radiologic and electromyographic findings supported this suggestion. The pain was demonstrated to be muscular tenderness, which was maximal and specific at motor points. Treatment directed to the cervical spine appeared to give relief in the majority of patients. The more resistant the condition, the more severe were the radiologic and electromyographic findings in the cervical spine.


Lymphoid interstitial pneumonia.

Lymphoid interstitial pneumonia. A 19-year-old man presented with dyspnea, cough and chest pains; he also complained of nausea, anorexia and postprandial vomiting and reported a 10-kg weight loss. Generalized lymphadenopathy and some rales over both lung bases were noted and a chest radiograph showed bilateral nodular lesions. Persistent leukocytosis, thrombocytosis, proteinuria and anergy to a series of natural antigens were found. The diagnosis of lymphoid interstitial pneumonia was made from material obtained at open lung biopsy. Rapid but incomplete clearing of the lung lesions resulted from steroid therapy; the other abnormalities were corrected gradually, except for the proteinuria, which persisted. The clinical improvement and the ability to work and play have been maintained for the past 20 months.


12-hour psychotherapy.

12-hour psychotherapy. The two cases cited illustrate how short-term therapy can be used to meet a patient's needs to deal with loss and grief. Although the goals as set out in short-term therapy contracts are not always fully achieved, this treatment modality, as delineated by Mann, is valuable to psychiatry. Short-term therapy is not only a sound therapeutic intervention, but also a means of bringing psychiatric health care to a greater segment of the population.


Effect of altered lighting regimens, time-limited feeding, and presence of Ehrlich ascites carcinoma on the circadian rhythm in DNA synthesis of mouse spleen. The objectives of the series of experiments described were (a) to determine whether there was a circadian rhythm in the incorporation of tritiated thymidine into DNA of the spleen in mice kept on a conventional light-dark cycle and fed ad libitum, (b) to study the effect that different light-dark cycles and a time-limited feeding schedule had on this circadian rhythm, (c) to ascertain what effect the presence of an 8-day Ehrlich ascites carcinoma (EAC) had on the rhythm in DNA synthesis in the spleen and what effect the EAC had on the circadian rhythm in the mitotic index in the corneal epithelium, and (d) to determine whether there was a circadian rhythm in the duration of life-span in mice bearing the EAC. A circadian rhythm in the incorporation of tritiated thymidine into DNA of the mouse spleen consistently was characterized by a peak during the nocturnal phase and a trough during the diurnal phase of the 12-hr light-12-hr dark cycle. In animals bearing an 8-day EAC, the rhythm in DNA synthesis in the spleen was phase-shifted, its wave form was changed, and the overall 24-hr mean was increased significantly. The phasing of the rhythm in EAC-bearing mice was not reproducible. This finding demonstrated that the presence of the EAC severely altered the natural rhythm in DNA synthesis in the spleen and resulted in a rhythmic pattern which was constantly changing. The presence of an 8-day EAC, however, had no effect on the amplitude, overall level, or the phasing of the circadian rhythm in the mitotic index in the cornea. A staggered light-dark cycle of 2 weeks duration did not completely phase-shift the DNA synthesis rhythm in the spleen but did completely phase-shift the rhythm in the mitotic index in the corneal epithelium. In mice subjected to a daily feeding period limited to 4 hr, the rhythm in DNA synthesis in the spleen, in both EAC-bearing and non-EAC bearing mice, was phase-shifted such that the peak occurred during the time of feeding and the trough occurred prior to the feeding period. The rhythm in the mitotic index in the cornea was not phase-shifted or altered in any way by the feeding schedule and thus remained fixed to the light-dark cycle. The DNA synthesis rhythm in the normal spleen demonstrated a phasing very similar to the circadian rhythm in the length of survival in mice challenged with EAC and the circadian rhythm in DNA synthesis in the normal thymus.


Enhancing effect of allopurinol on the induction of bladder cancer in rats by n-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide. The effects of allopurinol on the induction of bladder cancer by N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT), excretion of urinary tryptophan metabolites, hepatic nitroreductase activity, and the acid-soluble thiol content of liver and blood in weanling female Fischer rats were investigated. Four groups of rats were given normal diet or normal diet supplemented with 0.005% allopurinol, 0.188% FANFT, or 0.005% allopurinol-0.188% FANFT. Transitional cell carcinomas appeared in 3 of 30 rats (10%) at 15 weeks and in 7 of 44 rats (16%) at 20 weeks in the FANFT-treated group; the carcinomas appeared in 14 of 35 rats (40%) at 15 weeks and in 27 of 50 rats (54%) at 20 weeks in the FANFT-allopurinol-treated group. Growth rate was not affected by allopurinol and FANFT. Allopurinol alone caused no morphological change in the epithelial cells of the urinary bladder but decreased hepatic cytosol nitroreductase activity. FANFT alone had no effect on hepatic cytosol or microsomal nitroreductase activity but increased hepatic and blood acid-soluble thiol content. FANFT increased the urinary excretion of anthranilic acid glucuronide, kynurenine, acetylkynurenine, and 3-hydroxykynurenine and decreased indican and o-aminohippurate excretion. Allopurinol did not alter the effects of FANFT on the acid-soluble thiol content of liver and blood or the excretion of urinary tryptophan metabolites.


Cytometric analysis of neoplastic transformation of vertebrate cell populations. Two parameters of neoplastic transformation in spontaneously transforming fibroblasts were examined. One parameter, the predilection for rounding of cells at metaphase, was studied using time-lapse cinephotomicrography and fixed, stained preparations. Ten different cell lines were assayed, including established lines of hamster, rat, and mouse cells, and six rodent cultures of known neoplastic potential as determined by animal injection and tumor production. There was a close correlation between the assumption by cells of a spherical shape at metaphase and their ability to form tumors on injection in syngeneic hosts. The projected area of adherent cells 24 hr after plating over a coverglass was used to assay the transition of these rodent cell populations in culture from nonneoplastic to neoplastic. As the cell population became neoplastic, there was a significant decrease in the mean projected area of the cells. Furthermore, as the cell cultures became capable of producing tumors, the projected area profile of the population shifted proportionally to smaller area classes.


Effects of adriamycin on ultrastructure of nucleoli in the heart and liver cells of the rat. Ultrastructural studies of the effects of adriamycin on liver and cardiac cell nucleoli of the rat showed that nucleolar segregation occurred within 1 hr after an i.v. injection of a 40-mg/kg dose. Between 3 and 27 hr after this single dose, liver cell nucleoli progressively reverted to a normal ultrastructure. Nucleoli of rat myocardial cells did not recover but underwent further fragmentation, segragation, and conversion to ring-shaped structures. The ultrastructural alterations of myocardial cell nucleoli may represent an important aspect of adriamycin toxicity.


Effects of route and schedule of administration of high-molecular levan on the growth of AKR lymphoma. The route and schedule of treatment with high-molecular levan markedly influences its inhibitory effect on the growth of transplanted AKR lymphoma. Injections of levan into the site of the primary tumor were more effective in inhibiting tumor growth and preventing tumor-associated weight loss and mortality than were i.p. injections. Local levan injections inhibited metastatic spread only in mice treated from Days 0 and 2. Levan i.p. was more effective in inhibiting metastases in animals started on treatment 7 to 13 days after tumor inoculation than in animals in which levanization was started earlier. Local injection of levan before inoculation of tumor enhanced tumor growth and shortened life-span in comparison to nonlevanized animals. In mice treated with levan for a short period only, the inhibitory effect on tumor growth slowly vanished within 2 weeks. Some animals treated for 5 to 8 months remained completely free of tumor. The results indicate that the effect of levan on tumor development is  mainly topical and depends on the concentration of the polysaccharide in the site. The tumor growth period from 0 to 5 days appears to differ from the following period in the reaction to levan treatment. The nature of this difference is not clear, but possible explanations are discussed.


Tumor induction by a single subcutaneous injection of sterigmatocystin in newborn mice. Sterigmatocystin, a mycotoxin produced by Aspergillus versicolor, Aspergillus sydowi, Aspergillus nidulans, and a species of Bipolaris, was given to newborn BALB/c X DBA/2F1 (hereafter referred to as CD2F1) mice by a single s.c. administration in 1% gelatin suspension. In an acute toxicity study, the maximum tolerated dose of sterigmatocystin was 5 mug/g body weight. In a chronic study, a single s.c. injection of 5, 1, or 0.5 mug/g body weight gave rise to high incidences of lung and liver adenomas when the animals were killed at the end of 1 year. The incidence of both tumors in mice at the dose of 5 mug/g body weight was statistically significant, and the incidences of lung tumor in female mice and of liver tumor in male mice at the dose of 1 mug/g body weight were also statistically significant, compared with tumors in control mice. Other tumors also were induced in treated mice (two malignant lymphomas and one adenoma of the submaxillary gland), in contrast to a zero incidence in vehicle control mice. These results confirm that a small quantity of sterigmatocystin induces tumors of lung and liver and that the dose of sterigmatocystin is related to the incidence of tumors in mice.


Vasoformative sarcomas arising from BALB/3T3 cells attached to solid substrates. The BALB/3T3 mouse embryo cell line, noted for its marked postconfluence inhibition of proliferation, anchorage dependence, and high serum requirement, and frequently studied as a prototype nontumorigenic "fibroblast" line that is compared with tumorigenic sublines transformed with various agents, produced tumors within 2 to 3 months when an average of 3 X 10(4) cells were implanted s.c. attached to 1- X 5- X 10-mm polycarbonate platelets. Plastic platelets alone produced no tumors after 1 year of observation. The tumors, as well as others arising from implants of BALB/3T3 cells attached to 3-mm glass beads, were given the histological diagnosis of "vasoformative saroma" because the tumor cells frequently formed vascular channels. The vasoformative pattern and the results of specific staining for reticulin and collagen support the likelihood that BALB/3T3 cells originated from endothelial cells rather than from fibroblasts. That the tumors were derived from BALB/3T3 cells and not host cells was proved when tumors arising in BALB/c X C57BL/6 F1 hybrids were shown to be transplantable to BALB/c but not to C57BL/6 mice. The cultured tumor cells showed loss of both postconfluence inhibition of proliferation and anchorage dependence. Evidence of the induction of endogenous oncornaviruses was obtained in only one of four tumors tested. These tumors also exhibited tumor-unique transplantation rejection antigens. We conclude that BALB/3T3 cells are preneoplastic and give rise to different spontaneously transformed clones bearing unique tumor rejection antigens when implanted in vivo attached to a solid substrate.


Modification of the response to actinomycin D-induced sublethal damage by simultaneous recovery from potentially lethal damage in mammalian cells. Exponentially growing cells and cells that were allowed to enter a plateau or nondividing state of growth by depleting the medium of growth-essential nutrients were used in these studies. Nondividing Chinese hamster ovary cells in vitro are less sensitive to actinomycin D (AMD) than are exponentially growing cells. The cells were tested for their ability to recover from AMD-induced potentially lethal damage (PLD) or sublethal damage. The proliferating and the nonproliferating cells do not recover from PLD or sublethal damage, and they experience further reductions in survival when they are maintained under the medium conditions of their respective growth states. However, when dividing cells are placed in conditions suboptimal for growth (incubated in depleted plateau medium after treatment with AMD), they did exhibit increased survival. The PLD recovery was so great under these conditions that it masked the true response of the cells to fractionated doses of AMD. When adjustments were made for the PLD recovery, the resulting data indicated a slight but measurable increase in survival. Since AMD inhibits cell progression in all stages of the cell cycle, this increase in survival observed with fractionated doses of AMD may be due to true recovery from sublethal damage, although the movement of cells into less sensitive stages of the cell cycle between treatments cannot be ruled out.


Immune response of BALB/c X DBA/2F1 mice to a tumor allograft during pyran copolymer-induced tumor enhancement. The immune response of BALB/c x DBA/2 F1 mice to a transplantable Moloney leukemia virus-induced tumor allograft (MBL-2) was studied to determine the mechanism of pyran copolymer-induced tumor enhancement. The relative levels of humoral, lymphocyte, and macrophage response were followed chronologically by in vitro cytotoxic microassays using 51Cr-labeled target cells. Although pyran increased the titer of humoral cytotoxic antibody, levels of humoral factors capable of abrogating lymphocytoxicity were not enhanced. Furthermore, splenic lymphocyte-mediated cytotoxicity, although slightly diminished in pyran-treated mice, was not significantly affected. Macrophages harvested from allograft-bearing animals exhibited marked tumoricidal activity, which was augmented by pyran treatment. This macrophage-associated activity was specific for MBL-2 cells and not attributable to cytotoxins elaborated into the culture medium. Pyran slightly activated macrophages from nonsensitized mice to become cytotoxic for MBL-2 cells; activation was not T-cell dependent. However, strikingly fewer macrophages infiltrated the allograft in pyran-treated animals as judged by both histopathology and direct measurement. The defect in the migration or deposit of macrophages at the allograft site may have contributed to tumor enhancement.


Kinetic alterations induced by 5-fluorouracil in bone marrow, intestinal mucosa, and tumor. In order to provide a more rational approach to the design of chemotherapeutic programs involving 5-fluorouracil (5-FU), the kinetic effect of this drug on bone marrow, gastrointestinal epithelium, and tumorous tissues in mice was monitored by determining the rate of incorporation of labeled deoxyuridine (UdR) into DNA following doses of 15, 50, and 100 mg/kg i.p: In BALB/c x DBA/2 F1 (hereafter called CD2F1) mice bearing P1534 ascites tumor, the magnitude and duration of suppression of [3H]UdR incorporation into DNA were directly related to drug dose for both the normal and tumorous tissues. However, the impact of 5-FU was clearly greater upon the P1534 ascites tumor than on either normal tissue, so that, after a dose of 100 mg/kg, the tumor did not initiate recovery until Day 5, while duodenal mucosa and bone marrow returned to 50% of control levels of UdR incorporation at 2 and 3 days, respectively. In order to determine whether the duration of suppression of [3H]UdR incorporation, such as observed in the P1534 ascites tumor, correlated with sensitivity of Ehrlich ascites tumor to 5-FU, the effect of 50 and 100 mg/kg of 5-FU i.p. on this less 5-FU-sensitive tumor was determined. No difference was observed in the time course of suppression and recovery between Ehrlich ascites tumor and the duodenal mucosa and bone marrow of the G.P. Swiss host. In non-tumor-bearing CD2F1 mice, the toxicity of a 2nd dose of 100 mg/kg 5-FU i.p. was found to be predictably related to the kinetic effect of a 1st dose of 100 mg/kg on [3H]UdR incorporation in the bone marrow and duodenal mucosa. In the time period from 12 hr through Day 6, when [3H]UdR incorporation into DNA rose to a maximum, administration of a 2nd dose of 5-FU was associated with 95% lethality. In contrast, during the period between 0 and 12 hr that preceded recovery of UdR incorporation, a 2nd dose resulted in less than 40% lethality, and, in the period from Day 6 through Day 8, when UdR incorporation was declining, lethality of a 2nd dose also declined to less than 25% lethality. In addition, the antitumor effectiveness of a 2nd dose of 5-FU, 100 mg/kg i.p., was predictably related to the differential effect of the 1st dose of 100 mg/kg on [3H]UdR incorporation in the P1534 tumor as compared to the host bone marrow and duodenal mucosa. Administration of a 2nd dose of 5-FU was maximally effective on Day 7, at a time when [3H]UdR incorporation was declining in the host target tissues and increasing to a maximum in the P1534 ascites tumor. Thus, the present study has demonstrated that [3H]UdR incorporation into DNA may be used to monitor the differential effect of 5-FU on tumor versus normal proliferating host tissues and, where such differences exist, to provide an in vivo method of value in designing a schedule of optimum therapeutic benefit.


Depression of host versus graft immunity and stimulation of tumor growth following partial hepatectomy. Tumor growth was measured in inbred (C3H and C3Hf) and hybrid (C3H X C57BL/6F1 and BALB/c X C3HF1) mice after partial hepatectomy, sham hepatectomy, and hind limb amputation. Epithelial tumors (mammary carcinoma) and mesenchymal tumors (methylcholanthrene-induced and spontaneous tissue culture) were used in order to determine the tissue specificity of the tumor growth stimulation. Immunological parameters were defined by: (a) utilization of tumors that were either antigenic or nonantigenic, and (b) measurement of the host versus graft response in partially hepatectomized mice. Partial hepatectomy and the control operations were done on the same day as the tumor cell inoculation. All tumors, regardless of their tissue type or antigenicity, grew significantly better in partially hepatectomized mice as compared with the control mice. There was  a significant positive correlation between the magnitude of tumor growth stimulation and the degree of antigenicity of the tumor. Last, skin allograft survival was significantly prolonged in the hepatectomized mice. The results suggest that : (a) the stimulation of tumor growth in hepatectomized mice is not tissue specific as previously reported; and (b) the antigenicity of a tumor is not necessary for growth stimulation after partial hepatectomy.


Effects of prednisolone on the differentiation of mouse lung adenomas in culture. Explants of pulmonary adenomas, induced in mice by urethan, were cultured with or without prednisolone for 72 hr. After this time, the cuboidal epithelial cells of the tumor contained many more lamellar bodies than the nonsteroid control cultures. Incorporation of labeled palmitic acid into saturated phosphatidylcholine was not significantly increased in these short-term steroid-treated cultures. The results indicate that steroid enhances the maturation of cultured pulmonary adenoma cells into cells morphologically indistinguishable from type II alveolar epithelial cells of normal lung.


Increased growth rate of a benzo(a)pyrene-induced transplantable tumor in bursectomized chickens. A benzo(a)pyrene-induced transplantable tumor was used to study the effects of bursectomy and thymectomy on tumor development in recipients that were isogenic with the donor for the major histocompatibility locus. Surgical bursectomies and thymectomies were done in different groups of chickens on the day of hatching, with no adjunct treatment. The following day, the experimental and control animals were given 10(3) live tumor cells s.c. Tumor frequency and size were followed weekly. With the experimental protocol used, thymectomy did not significantly affect tumor frequency or growth. Bursectomy did not affect tumor frequency, but the transplanted tumors grew significantly more rapidly in bursectomized than in control animals. Thus, bursectomy and, consequently, the antibody-forming system influence not only virus-induced tumors, but also the development of a chemical carcinogen-induced transplantable tumor line. The data underscore the host-protective role of the antibody-forming system in oncogenesis.


Physicochemical alterations in the conformation of rat liver chromatin induced by carcinogens in vivo. Administration of methylating carcinogens such as methyl methanesulfonate (120 mg/kg), dimethylnitrosamine (5 mg/kg), or methylnitrosourea (80 mg/kg) to rats resulted in an increased ellipticity in circular dichroism spectra and in an enhanced ability to bind ethidium bromide in the liver chromatin. Although shearing of the chromatin preparations increased both the ellipticity and number of binding sites for ethidium bromide, the carcinogen-induced effects were noticeable whether or not chromatin was sheared. Although the doses of the 3 carcinogens used in these studies are equivalent in their ability to induce strand breaks in liver DNA at 4 hr, their effects on the induction of conformational changes in liver chromatin are different. For example, methyl methanesulfonate induced the minimum conformational changes in liver chromatin at 4 hr, whereas methylnitrosourea induced the maximum changes at 4 hr. Methyl methanesulfonate and dimethylnitrosamine, on the other hand, induced maximum changes at 3 days. The conformational changes induced by methyl methanesulfonate and methylnitrosourea, and not by dimethylnitrosamine, tend to be repaired by 14 days.


Test for carcinogenicity of metallic compounds by the pulmonary tumor response in strain A mice. The production of lung adenomas in strain A mice following multiple i.p. injections of 13 metallic compounds was investigated. A significant increase in the average number of lung tumors per mouse was noted following the administration of lead subacetate, manganous sulfate, molybdenum trioxide, and nickelous acetate. These four compounds can be considered as weakly carcinogenic for lung tumors in strain A mice.


In vitro labeling and gold activation autoradiography for determination of labeling index and DNA synthesis times of solid tumors. In vitro labeling and gold activation autoradiography were used to determine the [3H]thymidine ([3H]TdR)-labeling indices and DNA synthesis times for C3H/He spontaneous mammary tumors. Three variations of the [3H]TdR, [14C]thymidine ([14C]TdR) double-labeling method, together with double-emulsion autoradiography, were used to determine the DNA synthesis times (TS). Tumors labeled totally in vivo (in vivo-in vivo method) and tumors labeled with [3H]TdR in vivo and subsequently labeled with [14C]TdR in vitro showed similar TS values. DNA synthesis times for tumors determined totally in vitro by double labeling (in vitro-in vitro method) were significantly longer than those observed in vivo; however, identical samples subjected to Hypaque-Ficoll gradient separation after double labeling showed TS's similar to those found in vivo. Furthermore, the interval between [3H]TdR and [14C]TdR administration had no effect on TS estimates in vitro. Gold activation autoradiography was used in the present experiments to reduce autoradiographic exposure times. This method, together with in vitro labeling, permits [3H]TdR labeling index and TS determinations after 6-hr and 7-day exposures, respectively.


Identification of the products of mitochondrial transcription in the walker corcinosarcoma by the use of actinomycin D and ethidium bromide. The major RNA species present in the purified mitochondrial fraction of the Walker carcinoma were investigated in order to determine which of them are located in the mitochondria and coded by the organelle DNA. The subcellular distribution of these RNA's and the in vivo sensitivity of the transcription process to selective inhibitors were examined. Among the different species separated by polyacrylamide gel electrophoresis, only the 21 and 16 Se RNA's were found exclusively in the purified mitochondria, approximately Se being the S value estimated from the relative electrophoretic mobility of the RNA. A bifid peak observed in the 16-15 Se region was shown to be an artifact caused by the ribonuclease inhibitor, naphthalene disulfonate. Ethidium bromide at high doses inhibited the incorporation in vivo of 32P into 21, 16, and 4 Se RNA, but the nuclear transcription of cytoplasmic RNA was also inhibited to the same extent. No significant effect was observed at lower doses. In contrast, actinomycin D exerted a differential inhibition of the synthesis of 28 and 18 Se RNA from both the cytoplasmic and the mitochondrial fractions, practically without affecting the transcription of the 21 and 16 Se species. The incorporation of 32P into mitochondrial 4 Se RNA was also considerably more resistant to the drug than the synthesis of the cytoplasmic tRNA. It is concluded that the 21, 16, and Se RNA's are the only major discrete species transcribed from mitochondrial DNA present in the Walker carcinoma.


Tumorigenic activity of lead chromate.

Tumorigenic activity of lead chromate. Lead chromate was investigated for its carcinogenic potential in both rats and mice. Results show that this compound is a very potent carcinogen in rats when administered i.m. Sixty-four % of the animals treated developed malignant tumors at the injection site. Three renal carcinomas were also found after i.m. treatment with lead chromate. Since lead powder is a comparatively weak carcinogen in rats, whether given p.o. or i.m., it is suggested that the combination of lead and chromium (also weak carcinogen) accounts for the high carcinogenic activity of lead chromate in rats. Swiss albino female mice could not tolerate the same high dose level as did the rats; at the lower dose administered to the mice, no tumors were detected.


Dynamics of leukemic and normal stem cells in leukemic RFM mice. RFM mice spontaneously develop a myelogenous leukemia that is transplantable into nonleukemic RFM mice. On transplantation, hemopoietic stem cells from leukemic mice (L-CFU-S) will seed in the spleen and grow as discrete colonies, as will hemopoietic stem cells from normal mice (N-CFU-S). As the leukemic cells used in these experiments have 39 chromosomes and normal murine cells have 40, it has been possible to estimate the numbers of N-CFU-S and L-CFU-S in RFM mice at weekly intervals after these mice had been given i.v. injections of 10(6) leukemic spleen cells (spleen cells from preterminal leukemic mice). At each study time, splenic weights, peripheral blood counts, and nucleated cell counts and colony forming units (CFU-S) of marrow, spleen, and blood were assayed. The karyotypes of dividing cells from and the histology of the resultant spleen colonies were also studied. Two weeks after the injection of leukemic spleen cells, the number of CFU-S in the marrow had increased to 3 to 10 times normal, that in the spleen to 100 times normal, and that in the blood was markedly increased. Three weeks after injection, the number of CFU-S in the marrow fell from the peak level at 2 weeks, the number in the spleen rose modestly, and the number in the blood continued to be markedly increased. A normal distribution of erythroid, myeloid, and megakaryocytic colonies was obtained from CFU-S assayed 1 week after injection of leukemic spleen cells, but from CFU-S assayed 2 or 3 weeks after injection of leukemic spleen cells, the colonies formed were comprised almost exclusively of myeloid cells. From spleen colonies formed from marrow or spleen cells obtained 1 week after the injection of leukemic spleen cells, all karyotypes contained 40 chromosomes, whereas from spleen colonies formed from marrow or spleen cells obtained 2 or 3 weeks after injection of spleen cells, almost all karyotypes contained 39 chromosomes. In contrast, most of the karyotypes found in spleen colonies formed from the injection of blood cells even 3 weeks after injection of leukemic spleen cells contained 40 chromosomes. All colonies containing cells with 39 chromosomes, leukemic colonies, contained only myeloid cells. We conclude that L-CFU-S differentiate only into the myeloid series. Early in the course of the disease there is an increase in both N-CFU-S and L-CFU-S in the spleen and marrow. As the disease progresses, the numbers of N-CFU-S in both spleen and marrow decline and, during the final week of the illness, the number of L-CFU-S in the marrow declines. The CFU-S in the peripheral blood are predominantly of normal type, even late in the disease when N-CFU-S are rare in the spleen and marrow.


Effects of manganese on carcinogenicity and metabolism of nickel subsulfide. Nickel subsulfide, Ni3S2, alone or combined with manganese or chromium dusts, was administered i.m. to Fischer rats to study the effects of the metals upon Ni3S2 induction of sarcomas at the injection site. The incidence of sarcomas within 2 years after injection of Ni3S2 (1.2 mg) plus manganese (1.0 mg) was 7%, versus 77% in rats that received only Ni3S2 (1.2 mg), and 80% in rats that received Ni3S2 (1.2 mg) plus chromium (1.0 mg) (p less than 0.005). No local sarcomas occurred in rats that received the injection vehicle, or in rats that received manganese or chromium without Ni3S2. Admixture of manganese diminished the solubility of 63Ni3S2 in rat serum, serum ultrafiltrate, or water, in vitro. Admixture of manganese with 63Ni3S2 did not affect the mobilization or excretion of 63Ni in vivo, nor did it alter the acute pathological reactions to Ni3S2. 63Ni concentrations in ultrafiltrates of supernatant fractions of homogenates of injection sites averaged 2.8 (S. D. +/- 0.7) ng/ml at 5 to 6 months after injection of 63Ni3S2 (1.2 mg) plus manganese (1.0 mg), versus 5.4 +/- 2.0 ng/ml after injection of only 63Ni3S2 (1.2 mg) (p less than 0.02). This study demonstrates that admixture of manganese dust and Ni3S2 inhibits Ni3S2 tumorigenesis in rats, and reveals that manganese dust affects the subcellular distribution of 63Ni derived from 63Ni3S2, without influencing 63Ni kinetics as estimated by compartmental analysis.


The effect of modifiers of microsomal enzymes on chemical oncogenesis in cultures of C3H mouse cell lines. Two cell lines, both derived from the C3H mouse and each having different responses (oncogenic and cytotoxic) to polycyclic aromatic hydrocarbon oncogens, were studied with respect to their drug-metabolizing enzymes. The 10T1/2CL8 cells (a C3H mouse embryo fibroblastic cell line) were much more effective in converting 3-methylcholanthrene (3-MC) to 3-MC water-soluble metabolites, 3-MC phenols, and 3-MC-bound cellular macromolecules than were CVP3SC6 cells (a new line of C3H mouse adult ventral prostate fibroblasts). Basal aryl hydrocarbon hydroxylase activity was higher in 10T1/2CL8 cells than in CVP3SC6 cells, while the reverse was found for epoxide hydrase activity (using 3-methylcholanthrene-11, 12-oxide as substrate. 3-MC or benz(a)anthracene induced epoxide hydrase activity in both cell lines to about the same extent. 3-MC did not induce aryl hydrocarbon hydroxylase activity in CVP3SC6 cells. Aryl hydrocarbon hydroxylase activity was markedly induced in both cell lines by benz(a)anthracene and was slightly induced in 10T1/2CL8 cells by 3-MC. In a chemical oncogenesis cell culture system, transformation of 10T1/2CL8 cells mediated by 3-MC could be increased two- to threefold by treating the cell cultures with: either benz(a)anthracene, styrene oxide, cyclohexene oxide, or 1,2,3,4-tetrahydrona=phthalene-1,2-oxide; or with cyclohexene or 1,2-dihydrona-phthalene, alkene precursors of cyclohexene oxide and 1,2,3,4-tetrahydronaphthalene-1,2-oxide, respectively. When 10T1/2CL8 cells were treated with a combination of  benz(a)anthracene and cyclohexene, 3-MC-mediated transformation was increased 7.8-fold. CVP3SC6 cells that were not transformed by 3-MC or other hydrocarbon oncogens were transformed by a combined treatment with benz(a)anthracene, 1,2-dihydronaphthalene, and 3-MC.


Clinical evaluation of ftorafur (pyrimidine-deoxyribose n1-2'-furanidyl-5-fluorouracil). Ftorafur, a possible sustained-release formulation of 5-fluorouracil, was administered to 27 patients with metastatic cancers. The majority of patients had adenocarcinoma, most of which (60%) arose from the gastrointestinal tract. Ftorafur was given i.v. at doses ranging from 1 to 3 g/sq m/day for 5 days, repeated every 2 to 3 weeks. Gastrointestinal (68%) and neurological (17%) toxicities were the most common side effects encountered in this study and became dose limiting at doses greater than 2 g/sq m/day for 5 days. Myelosuppression (7%) was infrequent. Other toxicities included weakness (20%), chills and fever (8%), and phlebitis (1%). Of 24 evaluable patients, 4 (17%) responded (1 complete and 3 partial remissions). Responses were seen in 1 of 8 carcinomas of the lung, 1 of 5 carcinomas of the stomach, 1 of 3 carcinomas of the colon, and 1 of 1 carcinoma of the jejunum. The duration of response ranged from 4 to 58 weeks. The results of this study resemble somewhat those obtained with the laborious 5-day continuous i.v. infusion of 5-fluorouracil. Daily doses of 2 g/sq m for 5 days, repeated every 3 weeks, produced significant antitumor effect and tolerable toxicity.


Stimulation or inhibition of immune resistance against metastatic or local growth of a C3H mammary carcinoma. The effects of syngeneic tumor vaccine preparations on resistance against s.c. and pulmonary implants of a spontaneous C3H mouse mammary carcinoma have been compared during progressive primary tumor growth, after surgical cure, and during declining immune resistance following complete tumor removal. Concomitant antitumor immunity reached a higher level of effectiveness in the lungs than in s.c. tissue. The concomitant immune resistance in the lungs also remained effective longer than did s.c. resistance, which was the first to decline under the increasing antigen burden of a growing tumor implant. Resistance to challenge recovered quickly after the complete removal of a large tumor burden, but recovery was impeded if killed tumor tissue was reinjected too soon after tumor removal. If injections of killed tumor tissue were given 7 days after tumor removal, the completed recovery was not affected. If injections of killed tumor tissue were given 30 days after tumor removal, immune resistance to challenge, at this time much declined, was stimulated.


Inhibitory effect of Capsella bursa-pastoris extract on growth of Ehrlich solid tumor in mice. The treatment of ICR mice with i.p. injections (0.14 g/kg/day) of the extract of Capsella bursa-pastoris herb (Cruciferae) caused 50 to 80% inhibition of the solid growth of Ehrlich tumor cells that had been inoculated into the s.c. tissue of the animals. The tumor lumps in the treated mice showed multifocal necroses and the infiltartion of host fibrous tissue cells. Experiments were also performed to isolate and identify the active component for the antitumor action, and an acidic substance was isolated in crystalline form from the herb extract. This acidic substance was identified as fumaric acid and was effective in inhibiting the growth of Ehrlich solid tumor at a dose of 10 mg/kg/day. The 50% lethal dose (i.p.) of this acid was 266 mg/kg.


Demonstration of fibrin in early stages of experimental metastases. Tumor emboli were produced in lungs of Sprague-Dawley rats by i.v. injection of Walker 256 tumor cells into the tail vein. Tissues were examined by electron microscopy at periods from 30 sec to 72 hr after tumor injection. Two methods of conventional staining were used, in addition to immunoperoxidase techniques, with antifibrin antibodies produced in rabbits. Tumor cells accompanied by a platelet mass were seen in pulmonary arterioles at the earliest time period (30 sec). By conventional staining, small amounts of fibrin were detected within the platelet clumps by 5 min after inoculation. Periodicity indicating stable fibrin was not seen by this technique until 15 to 45 min. When peroxidase-labeled antibody was applied to tissue, sections showed fibrin-positive material at 30 sec, and periodicity of fibrin was detected by 5 min. Fibrin reached a maximum by both techniques at about 1 hr and disappeared, along with the platelets, at about 9 hr. When fibrinolysin was injected prior to the tumor cell inoculation, platelets and fibrin were either absent or present only in traces, and no stable fibrin was detected. These observations show that fibrin occurs very early in small amounts in association with tumor cell emboli, and is removed while the cells are still intravascular.