Changes in the distribution of intramembranous particles in hen erythrocytes during cell fusion induced by the bivalent-cation ionophore A23187.

Incubation of hen erythrocytes with Ca2+ and the bivalent-cation ionophore A23187 induced slight cell fusion in 1 h at 37 degrees C, and extensive fusion during a subsequent 15 min at 47 degrees C. Redistributions of intramembranous particles were observed, possibly involving interactions between Ca2+ and phospholipids, which are discussed in relation to molecular mechanimss of cell fusion.


Scanning electron microscopy in the evaluation of endothelial integrity of the fatty lesion in atherosclerosis.

The luminal surface of fatty lesions of atherosclerosis was viewed by scanning electron microscopy (SEM). Endothelial cells were outlined by staining intercellular junctions with silver and the aortas were fixed in situ at physiological pressure. When aortas were dehydrated by passage through organic solvents followed by critical point drying from liquid CO2, there was considerable disruption of the luminal surface and it was not possible to correctly interpret the morphological integrity of the endothelium. In contrast, simple air-drying of aortas, without solvent dehydration after fixation, allowed the integrity of the cell layer overlying the lesion to be evaluated. The success of this technique was attributed to the retention of arterial lipids during dehydration of the tissue.


Cell-mediated immunity in subacute sclerosing panencephalitis.

The cell immune response, in vivo and in vitro, was investigated in eight patients with subacute sclerosing panencephalitis. Although the disease had been diagnosed at different stages and despite the persistently high titers of humoral antibodies, a definite deficiency of cellular immunity, as indicated by skin reactions to common antigens and by migration inhibition factor to specific antigens, was found in all the patients. The cellular immune response of two patients treated with small amounts of transfer factor was temporarily restored by their clinical state did not improve.


Plasma potassium changes following suxamethonium and suxethonium in normal patients and in patients in renal failure.

Fifty-four healthy patients and 21 patients with chronic renal disease received either suxamethonium 100 mg or suxethonium 150 mg for neuromuscular blockade during endotracheal intubation. A significant increase in plasma potassium concentration was observed following the administration of suxamethonium. No increase occurred in patients who received suxethonium. It is concluded that suxethonium may be preferable to suxamethonium in patients who are at risk from an increase in the plasma potassium concentration.


Cerebral responses to the addition of nitrous oxide to halothane in man.

Cerebral responses to the substitution of 60% nitrous oxide for nitrogen during halothane anaesthesia (0.84%, end-tidal) were studied in four patients during surgery. The mean (+/-SEM) cerebral blood flow equivalent and internal jugular venous oxygen tension during halothane anaesthesia, 17+/-3 ml blood/ml oxygen and 41+/-2 mm Hg respectively, increased significantly to 45+/-3 ml blood/ml oxygen and 54+/-3 mm Hg following the introduction of nitrous oxide. On the withdrawal of nitrous oxide, the mean cerebral blood flow equivalent and internal jugular venous oxygen tension returned gradually to the control values. Cerebral perfusion pressure and blood-gas values, other than the internal jugular venous oxygen tension, did not change significantly. Marked slowing of the e.e.g. was observed following the addition of nitrous oxide to halothane, Upon the withdrawal of nitrous oxide the e.e.g. returned to the control pattern. These results indicate that cerebral blood flow was in excess of oxygen demand during nitrous oxide/halothane anesthesia in man.


The effects of droperidol and fentanyl on intracranial pressure and cerebral perfusion pressure in neurosurgical patients.

The effects of droperidol and fentanyl on the intracranial pressure (i.c.p.) and cerebral perfusion pressure (c.p.p.) were studied in eight anaesthetized normocapnic patients with intracranial space-occupying lesions. The infection of droperidol resulted in a small and not significant increase in i.c.p. from 24.0 to 27.2 mm Hg, while c.p.p. decreased from 75.9 mm Hg to 57.8 mm Hg, as a result of a decrease in systemic arterial pressure. The addition of fentanyl produced no change in i.c.p., but a further decrease in arterial pressure decreased c.p.p. from 60.4 mm Hg to 47.8 mm Hg. In four patients values of c.p.p. less than 40 mm Hg were obtained. C.p.p. was was increased by hyperventilation in all but one of these patients. It is concluded that droperidol and fentanyl should be used in patients with intracranial hypertension only if hypocapnia has been established and when the arterial pressure is normal or increased.


Studies on dimethyl tubocurarine in anaesthetized man.

Intravenous doses of 0.3 mg/kg dimethyl tubocurarine were required for consistent and adequate surgical relaxation in patients during nitrous oxide anaesthesia and produced virtually complete neuromuscular paralysis (96-100%) of the tetanic and twitch responses of the adductor pollicis muscle. The duration of action of the drug was prolonged--more than 3 h was required to attain 50% recovery from full neuromuscular paralysis. At this degree of recovery neostigmine was an effective antagonist. Determination of the tetanic tension ratios showed that tetanic fade developed during onset of paralysis and remained maximal until about 25% recovery of the tetanic contractions, when it disappeared raidly as recovery progressed. These doses caused no significant changes in arterial pressure and heart rate and this absence of cardiovascular side-effects is a desirable property of dimethyl tubocurarine. However, for many surgical procedures, an agent with a similar pharmacological profile but with a shorter action is desirable.


The effect of halothane and thiopentone on ventilatory responses mediated bythe peripheral chemoreceptors in man.

The activity and responsiveness of the peripheral ventilatory chemoreflex were assessed by the transient depression of ventilation following two breaths of oxygen in air-breathing subjects, and the differing times of onset of the ventilatory response to i.v. sodium bicarbonate in subjects breathing either air or oxygen. In patients premedicated with pethidine, in whom anaesthesia was induced with thiopentone, it was found that halothane, in an inspired concentration of 0.7-0.8%, reduced the activity and responsiveness of the peripheral ventilatory chemoreflex markedly. When halothane was discontinued and anaesthesia was maintained wtih intermittent injections of thiopentone (0.2 mg/kg/min) evidence of peripheral chemoreceptor activity and responsiveness returned.


The effect of positive end expiratory pressure on lung mechanics and arterial oxygenation after open heart surgery in young children.

The effect of positive end expiratory pressure (PEEP) during mechanical ventilation of the lungs was measured in 10 young children, aged 3 weeks to 30 months, who were being ventilated artificially after cardiopulmonary bypass surgery. Changes in the end-tidal lung volume were measured directly using a jacket plethysmograph, and were assumed to indicate change in functional residual capacity (FRC). Simultaneous changes in compliance and blood-gases were measured also. FRC was increased in all subjects, and in most this increase was accompanied by an increase in arterial oxygenation. There was no significant alteration in the mean values of dynamic or static compliance in the group, although dynamic compliance increased during PEEP in six patients. PEEP may be of value after cardiopulmonary bypass in young children when oxygenation is poor despite high inspired oxygen fractions. The possible mechanisms by which it increases PaO2 are discussed.


The effects of adding adrenaline to etidocaine and lignocaine in extradural anaesthesia II: Pharmacokinetics.

The addition of adrenaline to solutions of etidocaine or lignocaine for extradural administration resulted in significantly lower plasma koncentrations in both the arterial blood (14% decrease for etidocaine and 43% decrease for lignocaine) and venous blood (35% and 60% decreases for etidocaine and lignocaine respectively). The nett absorption over the first 4 h after administration, measured from the area under the plasma concentration-time curve, was decreased by the addition of adrenaline (18% and 30% decreases for etidocaine and lignocaine respectively). Both drugs were absorbed in a biexponential manner, having similar fast absorption rates but with etidocaine having a longer absorption half-life in the slow absorption phase. It is concluded that the addition of adrenaline reduces the fraction of the dose being absorbed during the first (fast) phase rather than influencing the absorption rate constants.


Hypocalcaemia during rapid blood transfusion in anaesthetized man.

In anaesthetized patients, administration of citrated whole blood for 5 min at controlled rates of 50, 100 and 150 ml/70 kg/min resulted in decreases in the calcium ion concentration (Ca2+) of 14, 31 and 41%, respectively. Ca2+ returned rapidly to the control values after termination of the transfusion. Reciprocal changes in serum citrate concentrations occurred, suggesting that the transient hypocalcaemia was a result of redistribution of citrate and hepatic or renal clearance from the vascular space. The total serum calcium concentration did not change significantly during rapid blood administration. Normal saline infusion at 100 ml/70 kg/min caused no change in Ca2+; however, plasma protein administration at this rate resulted in an 18% decrease in Ca2+, presumably as a consequence of the binding of calcium ions to anionic sites on plasma protein. Hypocalcaemia accompanying blood transfusion is a transient phenomenon, dependent on the total dose of citrate administered and the rate of infusion. Rational calcium replacement therapy during massive blood transfusion may now be based on direct Ca+ measurement.


Increased platelet aggregation responses to 5-hydroxytryptamine in patients taking chlorpromazine.

1 The aggregation response of platelets induced by 5-HT was greatly increased in psychiatric patients receiving chlorpromazine therapy when compared with normal volunteers and psychiatric patients not receiving chlorpromazine. 2 Platelet aggregation responses to ADP were normal during chlorpromazine therapy, but 5-HT induced aggregation was increased in rate and the typical transient reversible response was converted to an irreversible response in all subjects. This was usually indistinguishable from the ADP response. 3 When chlorpromazine therapy was stopped, plasma concentrations of chlorpromazine, monodesmethylchlorpromazine and chlorpromazine sulphoxide fell rapidly within one week, whereas 5-HT induced platelet aggregation responses became normal after three weeks. The enhanced responses returned when chlorpromazine therapy was re-instituted. 4 It is possible that platelet aggregation responses to 5-HT in vitro could prove to be a useful index of the pharmacological effect of chlorpromazine in vivo.


Resuspension of platelets: enhanced 5-hydroxytryptamine-induced aggregation in chlorpromazine treated patients due to changes in platelet properties.

1 Previous work showed that platelets from patients receiving chlorpromazine (CPZ) have an enhanced aggregation response to 5-HT. This was caused by an action of CPZ on the platelets, a factor borne by the plasma, or both. 2 Therefore we studied 5-HT induced aggregation of platelets from CPZ treated patients resuspended in normal plasma and from normal subjects resuspended in CPZ plasma. 3 Aggregation responses of normal platelets to 5-HT were not enhanced by resuspension either in normal plasma or in CPZ plasma. In addition the enhanced responses of platelets from CPZ treated patients did not revert to normal after resuspension in normal plasma. 4 It is concluded that the enhanced aggregation response to 5-HT seen after CPZ treatment is due to a change in the properties of the platelets rather than a factor in the plasma.


The effect of metoclopramide on the absorption of effervescent aspirin in migraine.

1 The absorption of effervescent aspirin was studied in two groups of patients during attacks of migraine. The first group received effervescent aspirin alone whilst the second group received intramuscular metoclopramide before effervescent aspirin. 2 After effervescent aspirin alone there was significant impairment in the rate of aspirin absorption during migraine attacks compared with the rate of aspirin absorption in normal volunteers and in the same patients when headache-free. 3 When metoclopramide was given before effervescent aspirin the rate of aspirin absorption during migraine attacks was not significantly different from that obtained in normal volunteers given effervescent aspirin alone or from that obtained in the patients themselves when given both metoclopramide and effervescent aspirin when headache-free. 4 It is concluded that the impairment of absorption of effervescent aspirin during migraine attacks is related to impaired gastro-intestinal motility with delayed gastric emptying and that this impaired motility can be overcome by parenteral metoclopramide.


Beta-adrenergic receptor blocking drugs, hypertension and plasma renin.

1 Propranolol and pindolol reduced both the blood pressure and plasma renin activity when given chronically to hypertensive patients. 2 There was no correlation between the fall in blood pressure and the fall in plasma renin activity. 3 Neither the basal nor the random plasma renin activity predicted the patients who would respond to beta-adrenergic receptor blocking drugs. 4 Oral propranolol reduced plasma renin activity but did not reduce blood pressure within 4 h of administration; oral pindolol reduced blood pressure but did not reduce plasma renin activity within 4 h of administration. 5 The reduction of blood pressure by propranolol and pindolol does not seem to be mediated by changes in plasma renin in most patients.


Interactions of orphenadrine and phenobarbitone with chlorpromazine: plasma concentrations and effects in man.

1 Two studies were carried out on acutely psychotic patients receiving chlorpromazine (100 mg) 8-hourly. 2 In the pilot study on five patients, plasma chlorpromazine concentrations fell over the course of 3 weeks of treatment and parallel changes were noted in the plasma half-life of antipyrine, salivation rate and handwriting length. 3 In the main study involving twelve patients treated for 15 weeks, the above findings were confirmed and were interpreted as indicating that chlorpromazine accelerated its own metabolism by inducing liver microsomal oxidising enzymes. No metabolites of chlorpromazine were detected in plasma. 4 The addition of phenobarbitone (50 mg) 8-hourly for 3 weeks, or orphenadrine (100 mg) 8-hourly for 3 weeks, resulted in a lowering of plasma chlorpromazine concentrations together with a further shortening of plasma antipyrine half-life. 5 Physiological effects of the additional treatments suggested that phenobarbitone lessens the effects of chlorpromazine by lowering body concentrations. However, orphenadrine acts more by virtue of its anticholinergic effects. 6 It was concluded that phenobarbitone and orphenadrine should not be prescribed routinely in patients receiving major tranquillisers. The need for the addition of orphenadrine should be assessed in each individual case.


Pharmacokinetics of N-demethyldiazepam in patients suffering from insomnia and treated with nortriptyline.

1 Hospitalized patients suffering from insomnia were treated with N-demethyldiazepam (30 mg p.o.) for 10 days, while continuing treatment with nortriptyline (75 or 100 mg daily). 2 Clinical evaluation performed by objective rating scales showed a positive therapeutic effect. No correlation was found however between drug plasma levels and clinical efficacy. A high inter-individual variability was observed. 3 In three cases the N-demethyldiazepam disappearance curve showed a biexponential decay, with a slow first component followed by a faster one. 4 Compared with diazepam the tested drug has a longer plasma apparent half-life and a lower relative clearance.


The effect of chloramphenicol and sulphaphenazole on the biotransformation of cyclophosphamide in man.

1 The metabolism of cyclophosphamide was investigated before and following treatment with chloramphenicol (five subject) and sulphaphenazole (seven subjects). 2 The biotransformation of cyclophosphamide was retarded after chloramphenicol treatment as indicated by prolongation in the mean serum half-life from 7.5 h to 11.5 h combined with decreased peak activity and integrated concentrations of metabolites. 3 In the sulphaphenazole treated subjects the rate of biotransformation of cyclophosphamide was significantly inhibited in two, unaltered in three and enhanced in two. The inconsistency of the results is discussed.


Assessment of drugs in schizophrenia. Diagnosis and patient selection.

It would seem that some set of operational definitions are required for the spectrum of psychotic patients often embraced by the rubric schizophrenia. The problems of acute (first-admission) and chronic patients have been described. At the present time 'relapse re-admissions' would seem to constitute the best population for drug evaluations. The importance of factors such as age, sex, ethnics, geography and length of history has been emphasized. Finally the importance of carefully excluding patients who would be at risk from pre-existing disease or hypersensitivity has been emphasized.


Haemodynamic effects of long-term oral labetalol.

1 The haemodynamic effects of labetalol have been studied in 11 hypertensive patients after 1 month of oral administration at a fixed dose of 800 mg/d. 2 Heart rate blood pressure and cardiac output were measured at rest and at two levels of exercise. 3 Labetalol significantly reduced heart rate and blood pressure at rest and after exercise but cardiac output remained unchanged. 4 The results suggest that labetalol is an effective anti-hypertensive agent acting by competitive adrenoreceptor blockade.


Double-blind trial of labetalol.

1 A new anti-hypertensive agent (labetalol) with alpha- and beta-adrenoreceptor-blocking properties has been assessed in 30 patients in a general practice. 2 Significant reduction in blood pressure and pulse rate occurred at a dose of 400 mg/d. A further significant reduction in blood pressure but not of pulse rate occurred at a dose of 800 mg/d. 3 After 4 weeks' treatment on the higher dose the mean fall in systolic and diastolic pressures compared with the pre-trial pressures was 36 and 24 mmHg, respectively. Side-effects were generally mild.


Experience with labetalol in hypertension.

1 The effect of labetalol has been assessed in an open clinical trial in 19 patients with sub-optimal control of blood pressure on other therapy, and in a placebo-controlled cross-over study. Most patients were taking a diuretic before the trial and they continued to do so during the trial. 2 In the open trial, labetalol successfully lowered blood pressure in most of the patients. Average daily dose was about 600 mg. The placebo-controlled trial confrimed the anti-hypertensive effect. 3 A clear postural hypotensive effect was seen. A few patients noted postural faintness as a side-effect, especially in a hot environment; but it was not troublesome. 4 There were few other side-effects. One patient experienced an increase in serum glumatic--oxalic transaminase (SGOT) and another an increasing anti-nuclear factor titre, but it is not yet certain that these were due to labetalol.


Controlled comparison of labetalol and propranolol in the management of severe hypertension.

1 Labetalol, a new hypotensive drug combining alpha- and beta-adrenoreceptor antagonist properties, has been compared with propranolol in the treatment of severe hypertension (blood pressure 190/115-249/139 mmHg) in a double-blind trial lasting 14 weeks. Additional diuretic therapy was given to both groups of patients. 2 Both drugs caused an effective reduction in blood pressure, bbut labetalol caused a greater fall in pressure in the standing position and after exercise. Two groups of nine patients have each completed the trial so far. Group average pressures for the last 3 weeks of treatment were: for labetalol 137/87 supine, 121/84 standing, and 117/78 mmHg after exercise; and for propranolol, 138/87 supine, 132/93 standing, and 133/94 mmHg after exercise. 3 Group average heart rates were lower in all three positions for those patients treated with propranolol compared with labetalol. 4 The average final dose ratio for labetalol: propranolol was 1.44:1 (w/w). 5 Labetalol initially induced a number of side-effects, predominantly related to alpha-adrenoreceptor blockade, which disappeared by the end of the trial. 6 Labetalol, in conjunction with diuretic therapy, was at least as effective as propranolol in lowering blood pressure in patients with severe hypertension.


Comparative anti-hypertensive effects of labetalol and the combination of oxprenolol and phentolamine.

1 The interaction of phentolamine with the beta-adrenoreceptor-blocking drug, oxprenolol, was studied in a controlled trial. 2 The combination was generally well tolerated. 3 Oxprenolol alone produced modest but significant reductions in supine, standing and post-exercise blood pressures. Small reductions were observed only with sustained phentolamine administration. 4 The combined effect of the two drugs seemed to be addictive only at the lower level of oxyprenolol dosage. 5 Labetalol produced significantly greater reductions in supine and standing blood pressure than combined oxprenolol-phentolamine. At a daily dose of 400 mg, postural hypotension was not observed, although transient symptoms were frequent.


Lung distensibility. The static pressure-volume curve of the lungs and its use in clinical assessment.

The main points of interest in the static expiratory PV curve are the changes in TLC (and to a lesser extent in RV), Pst(L) at standard volumes and particularly at TLC,and compliance (delta V/delta P) particularly close to FRC. More subtle changes in curvature may be present but have not as yet achieved any clinical or diagnostic significance. Although any presentation short of the whole PV curve inevitably conceals information a useful summary of the major changes can be obtained by considering only three points--the changes in TLC, Pst(L)max and compliance close to FRC (Fig. 22). In conditions associated with an increased TLC, four distinct patterns of change in the PV curve have been recognized resulting in different combinations of changes in Pst(L)max and compliance at FRC (Fig. 22, a, b, c, d). There are two main patterns of PV curve in restrictive lung disorders--one due to stiffening of the lung (Fig. 22, g, i, j) and the second due to extrapulmonary factors which prevent a normal distending pressure being applied to the pleural surface of the lung (Fig. 22, h). In practice it appears that lack of distending pressure leads to a secondary reduction in lung compliance. Nevertheless differences in Pst(L)max remain. The general patterns of abnormality may be summarized as follows: 1. Increases in TLC are almost always associated with a normal or increased compliance but Pst(L)max may be increased, normal or decreased. 2. Decreases in TLC are almost always associated with a decreased compliance but again Pst(L)max may be increased, normal or decreased. 3. When TLC is normal, it is theoretically possible that the whole PV curve may be displaced on the pressure axis and compliance may be altered (as in ageing) but such changes have not been identified in clinical practice.


Effect of blood pressure angiotensin II and aldosterone concentrations during treatment of severe hypertension with intravenous labetalol: comparison with propranolol.

1 The actions of labetalol 1.0-2.0 mg/kg intravenously on blood pressure, heart rate, plasma angiotensin II and aldosterone concentrations have been studied in 20 recumbent hypertensive patients. 2 In all subjects there was a reduction in systolic and diastolic blood pressures within 5 min of completion of injection. 3 Severe hypotension was not seen but three patients who had a marked fall in blood pressure experienced side-effects. Postural hypotension was common at the end of the study. 4 Labetalol caused significant reduction in heart rate. 5 Labetalol induced significant lowering of plasma angiotensin II and in plasma aldosterone concentrations, which were most obvious when these were increased initially; overall there was a close correlation between concurrent measurements of angiotensin II and aldosterone concentrations. 6 In five patients a comparison was made against propranolol 10 mg intravenously. Labetalol was more effective in lowering blood pressure but less effective in reducing pulse rate of plasma angiotensin II concentration.


Lymphokines (MIF) in the serum of patients with sarcoidosis and cutaneous granuloma annulare.

Serum MIF activity was studied in ten patients with sarcoidosis, fourteen with granuloma annulare, four with necrobiosis lipoidica, and nine with various dermatological diseases. Positive MIF activity was found in the sera of nine of the ten patients with sarcoidosis and eleven of the fourteen patients with granuloma annulare. The delayed hypersensitivity tests were negative in all nine of the patients with sarcoidosis who had serum MIF activity and were positive in only three patients with cutaneous sarcoid lesions. One of four patients with necrobiosis lipoidica demonstrated minimal serum MIF activity. Data on serum lymphokine activity in sarcoidosis and granuloma annulare suggest that these two diseases are related to delayed hypersensitivity mechanisms.


Granuloma annulare: direct immunofluorescence study.

Direct immunofluorescence studies were carried out in eleven specimens of granuloma annulare. The results (in all cases) indicate the presence of focal deposition of fibrin, localized primarily in the intervascular portion of the dermis, corresponding to the granulomatous and necrobiotic areas. These findings implicate the cellular mechanism of delayed hypersensitivity with focal involvement of the clotting system in the development of granuloma annulare lesion.


Localized pemphigoid.

Of 144 patients with bullous or cicatricial pemphigoid, nine with localized pemphigoid were seen at the Mayo Clinic between 1968 and 1975. In two patients the disease had become generalized before presentation, and in one it had evolved into bullous pemphigoid, the only case with positive indirect immunofluorescence. One additional patient showed mucosal lesions. Direct immunofluorescence was performed in five patients and showed linear deposition of C3 and fibrin but not immunoglobulins in three; in two patients immunofluorescence was negative. Localized pemphigoid can be divided into two types: that with scarring plaque-like lesions usually occurring on the head and neck predominantly in males, and that with localized bullous lesions usually occurring on the lower part of the legs of females.


Bullous pemphigoid and psoriasis: does subclinical bullous pemphigoid exist?

Four cases are presented in which probable bullous pemphigoid and probable psoriasis co-existed. One case represents a simple coincidence of the two diseases. In the other three cases, the bullous lesions were transient and seemed to be induced by elements of the Goeckerman regimen. The hypothesis is presented that these three cases represent a subclinical form of bullous pemphigoid induced by the irritant effects of coal tar, ultraviolet light, the reduced barrier function of a pre-existing dermatosis, or a combination of these factors.


Bullous dermatosis of chronic renal failure.

Four of five patients with chronic renal failure developed tense bullae on the backs of their hands and fingers. In one of these patients the bullae were more widespread. In the fifth patient the lesions were limited to the fingers. The clinical picture resembled that of porphyria cutanea tarda, but porphyrin studies were consistently within normal limits. Histological studies showed subepidermal pressure bullae in each patient. In one of the patients immunohistology showed greenish fluorescence at the bases of the dermal papillae.


Identification of human mononuclear cells by anti-monocyte serum.

An antiserum to human leukaemic monocytes was raised in rabbits. Adsorption with human thymocytes and chronic lymphatic leukaemia cells rendered the antiserum specific for cells of the human myeloid and monocytic cell series. Additional adsorption with granulocytes left a weak, but specific, activity for monocytes. The cells in normal peripheral blood with which the antiserum reacts by an immunoperoxidase technique have typical monocyte morphology. Cells with typical lymphocyte morphology do not react with the antiserum, but a few cells with intermediate morphology were also stained by the antiserum. Using the antiserum as a marker, the precursor cells of the monocytic and myeloid cell series were traced in the bone marrow.


Indium chloride scintigraphy: an index of severity in patients with aplastic anaemia.

[111In]Indium chloride scans of the bone marrow were performed in 22 patients with idiopathic aplastic anaemia before therapeutic intervention. In 20 patients there was a marked reduction in the uptake of indium chloride by the marrow, and in two patients the uptake of indium chloride was normal. Nineteen of the 20 patients with reduced marrow uptake of indium chloride, who underwent bone marrow transplantation, died or failed to improve. One of the two patients with normal scans improved and the other has remained stable. Four patients studied both pre and post successful bone marrow transplantation had minimal marrow uptake before transplantation and normal scans after transplantation. Failure of indium-111 choride marrow uptake correlates with poor prognosis in idiopathic aplastic anaemia.


The heterogeneity of erythrocyte antigen distribution in human normal phenotypes: an immunoelectron microscopy study.

A and A1 antigen were detected on human blood erythrocytes by immunoelectron microscopy using peroxidase-conjugated antibodies. Cells were obtained from various normal A subgroups, including rare weak A phenotypes and infant (cord blood) samples. Erythrocytes were fixed prior to incubation with specific reagents. The detection of surface antigens was carried out by an indirect method involving anti-A and anti-A1 antibodies and conjugated anti-immunoglobulin antibodies. The surface labelling was seen as a diffuse dense layer. Haemperoxidase-like activity resulted in a faint background which did not interfere at the level of ultrathin sections, with surface staining due to exogeneous peroxidase. The most significant finding was the existence, in a given sample, of several populations of cells as revealed by their antibody-binding capacity. The distribution of the various populations varied from one sample to another according to its subgroup. The progressive weakening of phenotype expression which characterizes the various subgroups from A1 to A weak was paralleled by a decreasing number of "antigen rich" cells, which were still detectable in weak phenotypes as a minor population. This study confirms that a given normal phenotype in fact represents a mixture of antigenically different populations of erythrocytes.


Differential expression of cell surface binding sites for cholera toxin in acute and chronic leukaemias.

Binding of purified cholera toxin to cell surface receptors has been visualized by an indirect immunofluorescence procedure. Normal nucleated cells from blood, bone marrow and lymphoid tissues, express these receptors with the possible exception of erythroid precursors. Cells from patients with chronic lymphoid or myeloid leukaemias have a normal receptor expression but acute leukaemic cells showed a marked deficiency in cholera toxin binding. Insertion of purified Gm ganglioside into membranes of acute leukaemic cells provided cellular binding sites for the toxin.


The effect of oral contraceptives on plasma cortisol and cortisol binding capacity throughout the menstrual cycle in normal women.

Plasma cortisol and cortisol binding capacity (CBC) was measured throughout the menstrual cycle in 25 healthy women aged between 20 and 35 years. Ten women were taking an oral contraceptive containing 50 mug oestrogen and progestogen ("combined pill"), one patient took a progestogen-only contraceptive and 14 served as controls. Despite the low oestrogen content of the combined pill the level of plasma cortison and CBC were signifcantly higher than in controls at all stages of the cycle, and the cortisol/CBC ratio greater than unity during most of it. Levels in the women taking a progestogen-only contraceptive were indistinguishable from controls. The plasma cortisol and CBC levels were significantly higher in the last week of the cycle than in the first in the combined pill group, but there was no significant variation in the controls.


Infertility and bilharziasis of the female genital tract.

In a prospective study, 138 African patients with infertility and 42 patients with incomplete abortion were examined for evidence of bilharziasis. There appeared to be a significant association between primary infertility and bilharziasis due to Schistosoma haematobium.


The effects of the fenoterol hydrobromide (Partusisten) aerosol on uterine activity and the cardiovascular system.

Fenoterol hydrobromide (Partusisten) aerosol was used in a placebo-controlled, double-blind, double crossover trial to study its effects on oxytocin-induced uterine contractions and the maternal cardiovascular system. The active and placebo aerosols were each administered a total of ten times or twice each in five patients who were in established oxytocin-induced labour with intact membranes. Five puffs, of 200 mug each, of the active aerosol reduced uterine activity by more than half; the effect on the cardiovascular system was to cause a mild tachycardia and to widen the pulse pressure.


Location of the protein hormones of the placenta by the immunoperoxidase technique.

An attempt has been made to locate human placental lactogen (HPL) and human chorionic gonadotrophin (HCG) in cells within the human term placenta. Using immunoperoxidase techniques on frozen sections of tissue, no difference could be found between specific antisera and control sera. A theory is put forward that it may not be possible to visualize these hormones in the placenta using present immunoperoxidase techniques.


The colposuspension operation for urinary incontinence.

Forty-two patients with urinary incontinence and related symptoms were treated by colposuspension. Two patients were lost to follow-up and the results of surgery in the remaining 40 patients are presented and discussed. The place of vaginal surgery in women with urinary symptoms is also discussed.


Mycotic keratitis in Nigeria. A study of 21 cases.

A clinical and mycological study of 21 cases of mycotic keratitis, a clinical entity not yet reported from Nigeria or West Africa, showed that Fusarium solani was the predominant aetiological agent. It was isolated from 12 cases. Four of the remaining nine cases were caused by Aspergillus fumigatus, one by A. flavus, two by Penicillium citrinum, and one each by P. expansum and Penicillium sp. All the 12 isolates of F. solani grew well at 37 degrees C and survived at 40 degrees C. Two cases, one due to F. solani and the other to A. fumigatus, were accompanied by panophthalmitis.


Keratometric changes after cataract extraction.

One hundred cases of uncomplicated cataract with clear corneae had cataract extraction and were examined at 20 days and six weeks after operation. Keratometry was done preoperatively and keratometry and retinoscopy at each of the follow-up visits. The study was undertaken to evaluate the type (whether pre- or postplaced) and the number of sutures (one, three, or five) that produce the least postoperative astigmatism. We found that the type of suture was immaterial but that three sutures produced the least astigmatism.


Cryptopleurine--an inhibitor of translocation.

Ribosomes from a cryptopleurine-resistant mutant of yeast were analyzed to determine the subunit localization of the resistance alteration. As was previously reported (Grant, P., Sanchez, L., and Jiminez, A. (1974), J. Bacteriol. 120, 1308), in vitro resistance of polyphenylalanine synthesis to cryptopleurine was conferred by 40S subunits from the mutant. Binding studies with sensitive ribosomes were carried out in order to identify the subunit binding site for cryptopleurine. Over the range of concentrations which inhibited polyphenylalanine synthesis, binding was proportional to concentration, so that a unique binding site could not be detected. Furthermore, binding to isolated subunits was about fourfold greater than to 80S ribosomes, suggesting that non-specific binding was sensitive to the condition of the particles. Model systems were developed in order to determine which step of the elongation cycle was inhibited by cryptopleurine. Elongation factor 1 dependent binding of Phe-tRNA to ribosomes was not inhibited by cryptopleurine concentrations, which inhibited polyphenylalanine synthesis. The initial rate of N-acetylphenylalanylpuromycin formation was inhibited when 10(-5) M cryptopleurine was added prior to translocation, but not when added after. Little inhibition was observed in either case when mutant ribosomes were used. These results suggest that cryptopleurine primarily inhibited translocation.


A cycloheximide sensitivity factor from yeast required for N-acetylphenylalanylpuromycin formation.

A protein (factor P) has been isolated from yeast, which was required for sensitivity to cycloheximide of a partially purified polyphenylalanine synthesis system. In the absence of factor P, 10(-3) M cycloheximide was required for 50% inhibition of polyphenylalanine synthesis, while in its presence, 10(-6) M gave 50% inhibition. Coincident with cycloheximide sensitivity was an activity required for EF-2 dependent N-acetylphenylalanylpuromycin (N-AcPhePuro) formation. Transfer of N-AcPhe to puromycin from the tRNA bound in the presence of 26 mM MgCl2 required factor P, as well as EF-2. Studies with antibody against EF-2 demonstrated that P factor was not required during the EF-2 translocation step but for some subsequent step.


Reactivity of ribosomal sulfhydryl groups in 30S ribosomal subunits of Escherichia coli and 30S-IF-3 complexes.

The reaction of 30S subunits with the SH group reagent N-ethylmaleimide (NEM) causes the loss of approximately 60% of their synethetic activity, but has little or no effect on the ribosomal binding of initiation factor IF-3. The ribosomal binding of this factor, on the other hand, was found to significantly influence the rate and the extent to which several 30S ribosomal proteins react with radioactively labeled NEM when the reaction kinetics of individual ribosomal proteins toward NEM were compared in 30S and 30S-IF-3 complexes. Of the nine 30S ribosomal proteins which react with NEM, proteins S1, S11, S12, and S18 were found to have lower reactivities, while proteins S4 and S21 displayed higher reactivity in the presence of IF-3. The reactivity of proteins S8, S13, and S17, on the other hand, appeared to be influenced little or not at all by the presence of the factor. These results are interpreted as supporting evidence for the premise that the binding of IF-3 results in a conformational change of the 30S subunit.


Menaquinone biosynthesis: conversion of o-succinylbenzoic acid to 1,4-dihydroxy-2-naphthoic acid and menaquinones by Escherichia coli extracts.

Two intermediates in the biosynthetic pathway to bacterial menaquinones are o-succinylbenzoic acid and 1,4-dihydroxy-2-naphthoic acid. Cell-free extracts have been prepared from Escherichia coli which catalyze the conversion of labeled o-succinylbenzoic acid to the naphthoic acid and also to menaquinones. The naphthoate synthetase has been partially purified and found to require coenzyme A and ATP. The synthetase has an approximate molecular weight of 45 000. The conversion of o-succinylbenzoic acid to menaquinones is stimulated by the presence of farnesyl pyrophosphate. The major menaquinone produced is then MK-3 with the farnesyl side chain.


beta-Galactosidase alpha complementation: properties of the complemented enzyme and mechanism of the complementation reaction.

Intracistronic alpha complementation involving Escherichia coli beta-galactosidase occurs between the cyanogen bromide peptide CB2, derived from residues 3-92 of beta-galactosidase (Langley, K.E., Fowler, A.V., and Zabin, I. (1975), J. Biol. Chem. 250, 2587), and the defective beta-galactosidase from the Z-deletion mutant strain M15. The M15 protein, a dimer, lacks residues 11-41 of beta-galactosidase (Langley, K.E., Villarejo, M.R., Fowler, A.V., Zamenhof, P.J., and Zabin, I. (1975), Proc. Natl. Acad. Sci. U.S.A. 72, 1254). The complemented enzyme formed from purified components has a molecular weight of 533 000+/-25 000, is therefore tetrameric, and has a probable stoichiometry of 1 CB2:1 M15 monomer. The complemented enzyme has the same Km for substrate as wild type enzyme, but is less stable to heat or urea treatment. The overall equilibrium constant for the complementation reaction is approximately 1-2 X 10(9) M-1. Initial velocity studies indicate saturation kinetics when either component is fixed and limiting, with an apparent Kd of about 10(-6) M. A first-order rate constant of 0.05-0.1 min-1 was estimated. The kinetics favor a model of rapid complex formation, followed by slow conformational change, as the mechanism of activation. Ultraviolet difference spectroscopy indicated an increased absorbance in the 290-300 nm region as a result of the complementation reaction. The kinetics of the increase suggest that two processes, one rapid and the other slower, could be responsible. The temperature dependence of complementation (Ea approximately 24 000 cal) is also consistent with the rate-determining step being a conformational change.


Separation of transfer ribonucleic acids on polystyrene anion exchangers.

The transfer RNA separation by chromatography on strong-base polystyrene exchange materials is examined and compared with the widely used reversed-phase chromatography. Results indicate important differences in some transfer RNA (tRNA) elution patterns by the anion-exchange chromatography, as compared with the reversed-phase chromatography. Transfer RNAs containing hydrophobic groups are adsorbed more strongly. The anion exchanger has twice the number of theoretical plates. Single peaks of tRNA2Glu and tRNA1Phe obtained from the reversed-phase column give multiple peaks only polystyrene anion-exchange chromatography. All six leucine tRNAs (Escherichia coli) and differences in tRNA populations synthesized during early and late stages of the dividing lymphocytes from normal human blood can be characterized by the anion-exchange chromatography. Different separation profiles are obtained by two separation systems for tyrosine tRNAs from mouse liver and mouse-plasma-cell tumor. The results indicate that, in contrast to the reversed-phase chromatography, strong-base-polystyrene anion-exchange chromatography is capable of separating tRNAs with minor structlral differences.


Equilibrium between two forms of the lac carrier protein in energized and nonenergized membrane vesicles from Escherichia coli.

p-Nitrophenyl alpha-D-galactopyranoside is a competitive inhibitor of lactose transport in membrane vesicles prepared from  Escherichia coli ML 308-225 (Ki congruent to 6.6 muM) but is not accumulated by the vesicles. Binding of p-nitrophenyl alpha-D-[6-3H]galactopyranoside to membrane vesicles has been measured by flow dialysis. In the presence of D-lactate, ligand binds to the vesicles with a KD of about 6 muM, and a total of 2.3 nmol per mg of membrane protein is bound at saturation. In the absence of D-lactate, a small amount of binding can be detected (approximately 0.2 nmol per mg of membrane protein) with a similar affinity constant (KD congruent to 9 muM). Binding inthe presence or absence of D-lactate is dependent upon a functional lac y gene product and upon the structural integrity of the vesicle membrane and is reversed by p-hydroxymercuribenzenesulfonate. Agents such as 2,4-dinitrophenol, carbonyl cyanide m-chlorophenylhydrazone, and valinomycin, alone or in combination, abolish D-lactate-dependent binding but do not affect binding in the absence of electron donors. The results confirm previous observations that the bulk of the lac carrier protein is unable to bind ligand unless the membrane is energized. and they also corroborate observations that a small amount of binding occurs in the absence of energy coupling. The findings are discussed in terms of a model in which the lac carrier protein exists in a state of dynamic equilibrium between two forms: (i) a low affinity, cryptic form which predominates in the absence of energy coupling; and (ii) a high affinity form, accessible from the external surface of the membrane, which predominates in the presence of an electrochemical gradient of protons (interior negative and alkaline).


Physical characteristics of protein-deficient derived from the 50S ribosomal subunit.

Sedimentation coefficients, diffusion coefficients, density increments, extinction coefficients, and molecular weights were determined for selected core particles from the 50S Escherichia coli ribosomal subunits. These core particles were found to be loosened structures containing less than half the protein present on the intact 50S subunits and totally lacking peptidyl transferase and polyphenylalanine synthesis activity unless reconstituted. The results of this study confirm that a loosening of the subunit takes place upon the removal of protein, but this loosening is not necessarily due to the loss of the protein.


Study on the surface polypeptides isolated from human lymphoblastoid ce-ls by ionic shock.

Ionic shock treatment in the presence of 10% glycerol is an efficient and selective method for extracting cell surface components from Raji cells and effecting the solubilization of up to 22 polypeptides. The majority of these shock-released polypeptides are accessible to lactoperoxidase radioiodination. Sera from rabbits immunized against these soluble extracts are reactive against Raji cell surface as indicated by indirect membrane immunofluorescence and agglutination assays.


[Fatty acid composition and phospholipid pattern in auxotrophs for unsaturated fatty acids (author's transl)].

The relationship between fatty acid composition and phospholipid pattern has been studied in Escherichia coli auxotrophs for unsaturated fatty acids. 1. The presence of a regulatory mechanism which enables the organism to maintain a given fluidity of the lipids has been corroborated using exogenous fatty acids which cause dramatic changes in fatty acid composition. 2. The fatty composition of phosphatidic acid is different from that of the other classes of phospholipids. 3. Changes in fatty acid composition are concomittant with the alteration of the phospholipid pattern. The ratio of phosphatidylglycerol to diphosphatidylglycerol is particularly sensitive to the physical characteristics of the exogenous unsaturated fatty acid. The relative increase in diphosphatidylglycerol is associated with membrane alterations.


Preparation of Escherichia coli pyruvate oxidase utilizing a thiamine pyrophosphate affinity column.

An improved procedure is reported for the purification of Escherichia coli pyruvate oxidase (pyruvate:ferricytochome b1 oxidoreductase, EC 1.2.2.2), a peripheral membrane flavo-enzyme, which is much more reproducible and requires considerably less time than the original purification scheme. The key element in this protocol is a new Sepharose-based affinity resin designed for the isolation of thiamine pyrophosphate-requiring enzymes. The synthesis, partial characterization, and use of two such affinity resins is described. Pyruvate oxidase is a pure, homogenous protein as it is eluted from the affinity resin. The enzyme is a tetramer with a subunit molecular weight of approx. 60 000. The subunits appear to be identical. The isoelectric point of pyruvate oxidase is 5.6.


Analysis of the kinetics of photoinduced crosslinkage of the protein and RNA components of the Escherichia coli 50 S ribosomal subunit.

The kinetics of photoinduced crosslinkage of the protein and ribosomal RNA components of the Escherichia coli 50 S ribosomal subunit were determined by two-dimensional gel electrophoresis. One group of ribosomal proteins was found to be reactive in the crosslinkage reaction at doses of 254 nm radiation of 2.9 X 10(3) ergs-mm-2. A second group of 50-S ribosomal proteins was found to be unreactive in the crosslinkage reaction at doses of 254 nm radiation less than 8.6 X 10(6) ergs-mm-2, but were reactive at higher doses.


Rifampicin binding as a probe for subunit interactions in Escherchia coli RNA polymerase.

The binding of the inhibitor rifampicin to RNA polymerase (alpha2betabeta') and its deficient subunit mixtures was investigated. The ability of beta to bind stoichiometric amounts of rifampicin was restored by formation of the alpha2beta subassembly. beta,beta' alpha, betabeta' and alpha2beta' were unable to bind rifampicin. RNA polymerase denatured with 6 M guanidine hydrochloride and dialysed against a renaturing buffer at 0degrees C ("renatured inactive enzyme") bound stoichiometric amounts of rifampicin but had lost the ability of bind dna. compared with native RNA polymerase "renatured inactive" enzyme possessed a markedly different tertiary structure as judged by limited proteolysis.


Structure, synthesis, and post-transcriptional modification of ribosomal ribonucleic acid in Bdellovibrio bacteriovorus.

The structure, synthesis, and post-transcriptional modifications of 23-S and 16-S ribosomal RNAs (rRNAs) have been studied in the facultatively parasitic bacterium, Bdellovibrio bacteriovorus. The mature 23-S and 16-S type of rRNAs in Bdellovibrio are larger than the analogous molecules in Escherichia coli by at least 1.0 - 10(5) and 0.5 - 10(5) daltons, respectively, and have a conformation different from E. coli rRNAs as judged by relative electrophoretic mobilities in polyacrylamide gels with and without denaturing conditions. Studies on the kinetics of synthesis and maturation of ribosomal RNA in Bdellovibrio show that precursor forms analogous to p23-S and p16-S in E. coli are synthesized. In addition, some earlier precursor rRNAs in Bdellovibrio are seen that appear analogous to the 25S and 17.5-S pre-rRNAs that have only been observed in the RNAase III deficient mutant of E. coli strain AB301-105 (Nikolaev, Birenbaum, M. and Schlessinger, D. (1975) Biocheim, Biophys. Acta 395, 478-489). These early precursor stages have not been observed in other procaryotic species, including E. coli that have normal levels of RNAase III. The results from the Bdellovibrio system provide that the 25-s and 17.5-S pre-rRNAs are normal stages of rRNA modification and are part of a multiple step maturation process, and therefore are not aberrations associated with the RNase III deficient mutation.


The synthesis and enzymatic polymerization of 5-thio- and 5-methylmercurithio-pyrimidine nucleotides.

The 5-thio and 5-methylmercurithio derivatives of UTP, dUTP and dCTP have been synthesized and tested as substrates for nucleic acid polymerases. The 5-thio-nucleotides were polymerized inefficiently by both RNA polymerase and DNA polymerase I of Escherichia coli. The 5-methylmercurithio derivatives of dUTP and dCTP were, however, utilized by DNA polymerase I, an enzyme insensitive to mercurial compounds, although they were potent inhibitors of all other polymerases tested. While polymers containing the 5-thio substituent possess structural abnormalities, most likely interstrand disulfide bridges, polymers containing 5-methylmercurithio groups appear normal. The latter polynucleotides are readily separated from non-sulfated polymers by chromatography on mercuriagarose.


General and specific effects of amino acid starvation on the formation of undermodified Escherichia coli phenylalanine tRNA.

The heterogeneity of undermodified phenylalanine tRNA produced in relaxed control E. coli during amino acid starvation was investigated. Examination of the RPC-5 elution profiles of tRNAPhe prepared from non-starved cells and cells starved of a variety of amino acids, including some known to be involved in the formation of modified bases revealed that: (1) only one species of fully modified tRNAPhe appears to occur in cells grown in enriched medium; (2) at least two chromatographically unique isoacceptor species are observed in addition to the normal tRNAPhe in starved cells; (3) the unique, undermodified species of tRNAPhe from leucine-starved cells, known to be deficient in dihydrouridine, pseudouridine, 2-thiomethyl-N6-(delta2-isopentenyl) adenosine and 3-(3-amino-3-carboxypropyl) uridine, co-elute with the unique species produced in cells starved of histidine or arginine or treated with puromycin or chloramphenicol; (4) additional unique species of tRNAPhe can be detected in methyl- and sulfur-deficient tRNA from methionine- and cysteine-starved cells; (5) analysis of phenoxyacetylated tRNA revealed that the chromatographically unique and normal species from starved cells contain subspecies deficient in 3-(3-amino-3-carboxypropyl) uridine; and (6) using phenoxyacetylation as a means of effecting the resolution of undermodified subspecies, a total of at least ten chromatographically unique subspecies of rRNAPhe were detected in an organism that appears to posses only one gene for tRNAPhe. Taken together, the results support the view that there are both general and specific effects of amino acid starvation on the post-transcriptional modification of tRNA.


Effects of an EMG biofeedback relaxation program on the control of diabetes: a case study.

It was hypothesized that EMG biofeedback relaxation training, applied to a diabetic patient, would result in a decreased level of insulin with fewer episodes of ketoacidosis. A 20-year-old female, diabetic since age nine, kept daily records of insulin doses and rated herself on an emotionality and a diabetic scale. A full-semester baseline was taken. This was followed by a semester-long training program during which the patient practiced relaxing her frontalis muscle with a portable EMG feedback unit which produced a geiger-counter-like click feedback. A cassette-tape series was used along with the portable EMG. The patient was encouraged to practice twice each day and to attempt to maintain a relaxed state even when not in the practice situation. The daily use of the portable unit was terminated at the end of the semester. In addition, the patient ceased practicing twice daily with the cassette tape. Daily insulin averaged 85 units for the six-week baseline and 59 for the final six weeks of the training period. Moreover, at the end of the training period the average dose had reached 43 units. During the training period the patient rated herself as decreasing in emotionality and in diabetic fluctuations.


Tension headaches: what form of therapy is most effective?

In order to assess the effectiveness of verbal relaxation instructions and EMG relaxation training on the relief of tension headaches, 18 medically documented tension-headache sufferers were randomly assigned to one of three conditions: (1) Jacobson-Wolpe autogenic-relaxation training, (2) EMG relaxation training, or (3) EMG relaxation training combined with Jacobson-Wolpe autogenic instructions. The two EMG-assisted groups showed significantly better results, compared to the verbal group, in terms of reduction of headache activity and the rate at which reduction took place. Thus, EMG-assisted relaxation procedures seem the treatment method of choice for relief of tension headaches.


Biofeedback and progressive relaxation treatment of sleep-onset insomnia: a controlled, all-night investigation.

Previous research suggests that self-defined insomniacs are distinguished from normals by high levels of anxiety and physiological arousal, which might be mitigated by muscle relaxation. This study assessed the relative effects of frontal EMG biofeedback, progressive relaxation, and a placebo set of "relaxation" exercises on the sleep of 18 onset insomniacs. Each subject was trained in one of these three methods for six half-hour sessions and slept in the laboratory for two consecutive nights before and after training. The experimental groups demonstrated significant decreases in physiological activity during training while changes in the control group were minimal. Reductions in sleep-onset time were: biofeedback group, 29.66 minutes; progressive relaxation group, 22.92 minutes; control group, 2.79 minutes. The experimental groups improved significantly (p less than 905) more than the control group, but did not differ from each other. No significant relationships between physiological levels and sleep-onset time were found, which suggests that muscle relaxation alone was not responsible for subjects' improvements. Since 20 minutes of daily practice were required to achieve an approximate 30-minute decrease in sleep-onset time, the practical utility of the methods is questioned.


Studies on the immunogical effects of BCG and its components: theoritical and therapeutic implications.

Bacillus Calmette-Guérin (BCG) has had significant antitumor effects in rodents and in man. In mice, BCG is capable of increasing lymphocyte-mediated immunity to allogenic leukemia cells and of creating "pseudoimmune" cytotoxic lymphocytes when administered alone. Lymphocyte-activating factor (LAF) produced by macrophages, a T-lymphocyte mitogenic substance, is increased significantly by the administration of BCG and may play a role in the evocation of cytotoxic cells. BCG acts as a mitogen for splenic and thymic cells "in vitro". Macrophages were important regulators in the response, since a critical concentration (at least 0.25 % to 0.5 %) were required for T-cell stimulation and a high concentration (approximately 5 to 10 %) inhibited splenic lymphocytes. LAF was increased 20-fold above baseline "in vitro" by BCG. Methanol extraction residue (MER) stimulated thymic and splenic cells as well as did whole BCG. A delipidated cell fraction (DMC) stimulated only splenic non-T lymphocytes. BCG has antagonized several sorts of immunosuppression and may be useful clinically in this regard. There is also the possibility of antagonizing the myelosuppressive toxic effects of chemotherapeutic agents through the stimulation of the macrophage-made granulocyte colony stimulating factor. Finally, if BCG can induce cytotoxic lymphocytes (or macrophages) in vitro, it may be possible to use these in clinical adoptive immunotherapy. BCG is a useful prototype of an immunological adjuvant with antitumor activity. Elucidation of its actions on lymphoid cells and its effects in therapeutic situations can serve as a model for future investigations with other substances with similar potential.


The computer-assisted morphological classification of acute leukemia. I. Preliminary results.

The morphology of leukemic lymphoblasts, as seen in the initial routine bone marrow aspirate, of 40 children with ALL has been evaluated using a computer-assisted automated microscope. A statistic, derived from a  combination of the number of macrolymphoblasts and the percentage of cells with no cytoplasm was found to predict response to standard therapy and to identify patients with a high probability of long-term remission as well as a group with a relatively poor response to therapy.


[GABA-ergic mechanism of action of diazepam on cerebral cortex neurons].

The effect of diazepam, depakin, thiosemicarbazide (TSC) and bicucullin on the spontaneous and incuced (by stimulation of the sciatic nerve) activity of the cells of the sensory-motor cortex was studied on the unanesthetized immobilized (curarized) albino rats. Diazepam proved to intensify the inhibitory phenomena, i.e. decreased the frequency of spontaneous discharges and prolonged the postactivation depression of the neuron impulsation (the inhibitory pause. Depakin increasing the GABA content in the brain acted similarly. The GABA-ergic receptors blocker, bicucullin and TSC, decreasing its content, on the contrary, weakened the inhibitory phenomena. There was also revealed a reversible antagonism between diazepam and bicucullin in their effect on the neuronal activity. The data obtained confirmed the supposition put forward formerly on the capacity of diazepam to intensify the effect of GABA on the neurons of the cerebral cortex.


[Use of cross-reacting microbial antigens reversal of tolerance acquired for mouse transplantation antigens].

Immunization of CC57BR mice tolerant to the C. albicans antigens and the affiliated transplantation antigens of C3H mice, with group A streptococcus (type I) containing the antigens cross reacting with the donor tissues, abrogated the tolerance to the C3H mice alloantigens. This was expressed in the shortened survival of the skin allotransplant and in increased rate of elimination from the lymph nodes and the spleen of labeled 51Cr donor lymphoid cells. The immune response to the C. albicans antigens was not restored.


[Independent integration of genes controlling the invasive properties and streptomycin resistance of enteropathogenic escherichiae 0124].

Recombinations which lost their invasive properties were obtained by crossing E. coli K12Hfr AB313 with entropathogenic escherichia belonging to serological group 0124. The loss of invasive properties by these recombinations was not connected with the acquisition of genes controlling the streptomycin resistance.


[Relationship between tumor antigens and histocompatibility antigens at the cell membrane].

The authors outline schematically the major histocompatibility complex as well as the relationship between tumor neo-antigen and pre-existing antigens at the cell surface. They note that these two antigenic systems are not independent and in particular, the major histocompatibility system (H-2) co-segregates with the tumour antigens. Considering the complexity of the H=2 system (a H-2 allele does not correspond to any single transplantation antigen but to a combination of several antigens units simultaneously present), the authors recall Boyse's hypothesis, revised by Haywood and Mc Khann, which propose that tumour antigens are a rearrangement of H-2 substructures. Moreover, it is possible that the relationship between H-2 and resistance to cancer may be attributed directly to the action of genes controlling the immune response (Ir region) which could also intervene in recognition of these tumour antigens. Finally, recent results obtained seem to show that some tumour antigens cross-react with H-2 fractions. This fact prevents the mice bearing these types of alleles from being immunized against the cross-reacting tumours. If this result could be transposed to the human situation, it would explain the frequency of certain types of tumours in defined H-LA groups and could allow the prediction of high-risk groups.


Problems in the use of in vitro assays for monitoring cell-mediated immunity in cancer patients.

Considerable controversy has evolved about the relevance of microcytotoxicity assays (both visual methods and isotope release assays) in the studies of cell-mediated immunity in human tumor systems. A stage has been reached, where technical aspects of these assays, capable of influencing the results, need to be considered: tumor target cell selection (the use of cells from short term cultures or cell lines), lymphocyte isolation procedures (composition of the effector cell populations) and selection of baseline controls for expressing the results, are recongnized as potential variables influencing results of in vitro cell-mediated cytotoxicity assays.


Bacteriological studies of contamination of hydrophilic contact lenses with Staphylococcus epidermitis.

An attempt was made to contaminate hydrophilic contact lens material with Staphylococcus epidermitis. This study was initiated as a result of some clinical investigations which had revealed the presence of coagulase-negative staphylococcal infection in eyes fitted with this type of contact lens. It was though particularly relevant in view of the known association of some strains of this organism with other plastic prostheses. In vitro cultures proved unsuccessful in demonstrating the ability of this bacterium to metabolize the lens material.


Diencephalic stimulation and the effects of ECT in endogenous depression.

We compared conventional bilateral ECT treatment electrode placement with simultaneous unilateral electrode placement to both sides of the head (dominant/nondominant unilateral ECT) in 20 patients with endogenous depression. Under double-blind random assignment conditions we found that six bilateral ECT were significantly more effective than six dominant/nondominant unilateral ECT in reducing depression rating scale scores. In the light of data from intracerebral ECT current distribution studies we interpret our findings to support our previously advanced hypothesis that diencephalic stimulation is requisite for the therapeutic benefit of bilateral ECT in endogenous depression.


Isolation and preservation of islets from the mouse, rat, guinea-pig and human pancreas.

Viable islets of Langerhans have been isolated from the mouse, rat, guinea-pig and human pancreas using a free hand microdissection procedure. Viablity has been assessed by light microscopy of sections stained with Gomori's aldehyde fuchsin and by measuring the insulin release from islets in vitro in response to a glucose stimulus. Ten pieces of human cadaver pancreas have been studied. Islets were isolated from 6 and in 5 cases were shown to respond to a glucose stimulus in vitro. Five pieces of human pancreas removed at operation have been studied. Islets were isolated in all cases but only 2 showed a response to a glucose stimulus. Isolated animal islets have been subjected to three preservation systems and their viability following storage noted. 1. Simple cold storage in Hank's balanced salt solution at 4 degrees C. At 15 hours 100% survival was noted. This dropped to 10% at 48 hours. There were no survivors at 72 hours. 2. Subzero storage. In group I (freezing rate 1 degree C/min) histological survival was 35% and functional survival 20%. In group II (freezing rate 5 degrees C/min with a 24-hour culture period after rewarming) histological survival was approximately 87% and functional survival 75%. 3. Organ culture. Islets from the guinea-pig, rat and mouse showed minimal morphological damage when cultured for 21 days in a simple organ culture system. At 28 days histological survival was approximately 30%. We were unable to correlate histological and functional survival in this group.


Endotoxin, bile salts and renal function in obstructive jaundice.

It is generally agreed that there is an increased incidence of postoperative renal failure in patients with obstructive jaundice. The proposition that this may be due to endotoxin, and that the endotoxin may be absorbed from the patient's own bowel flora, has been investigated. The study was conducted in patients and in animals. Twenty-four patients with obstructive jaundice were studied. Sixteen had endotoxin in the portal blood at operation and 13 of these had peripheral endotoxaemia at the time of operation or developed it during the 72 hours following operation. Only 3 of the patients had a proved possible site of infective origin for the endotoxaemia, the remainder absorbing the endotoxin from their own gastro-intestinal organisms. Portal endotoxaemia occurred when the serum bilirubin was 8-5 mg/100 ml or greater. There was a highly significant decrease in mean endogenous creatinine clearance postoperatively in patients with endotoxaemia, and expression of this as a percentage postoperative fall compared with preoperative levels enhanced the significance. Jaundiced patients without endotoxaemia had no significant fall in endogenous creatinine clearance. Matched non-jaundiced control patients did not develop portal or peripheral endotoxaemia, and there was no significant fall in postoperative creatinine clearance. Experiments using animals showed that the absence of bile salts in the intestinal tract in obstructive jaundice allows endotoxin to be absorbed, and that this absorption can be prevented by the oral administration of bile salts. The therapeutic implications in patients are discussed.


Layer closure of laparotomy wounds with absorbable and non-absorbable suture materials.

Opinions vary regarding the best suture material for abdominal wound closure. Some surgeons favour non-absorbable sutures while others use absorbable materials, but there is little objective clinical information regarding the relative merits of the different suture materials. In a randomized prospective clinical trial the synthetic absorbable polyglactin and polyglycolic acid sutures were compared with the non-absorbable polypropylene suture in the closure of 161 laparotomy wounds. A layered closure technique was used in all cases. The total incidence of wound dehiscence and herniation was 8-1 per cent, and there was no significant difference between polylycolic acid (9-6 per cent), polyglactin (5-8 per cent) and polypropylene (8-8 per cent). Dehiscence and herniation were significantly more common in wounds closed by surgeons in training. It was concluded that none of the suture materials provided satisfactory freedom from wound complications. It is possible that these sutures may prove satisfactory with an alternative technique of abdominal wound closure, but this should be the subject of further clinical studies.


Streptokinase in acute myocardial infarction: a controlled multicentre study in the United Kingdom.

In a multicentre trial of streptokinase in acute myocardial infarction 302 patients received an intravenous infusion of 2 500 000 IU of streptokinase over 24 hours, while 293 patients served as controls. Neither group received anticoagulants unless indicated by thromboembolic complications. No significant difference in mortality was evident during inpatient treatment nor at six-week or six-month follow-up. The inpatient death rate was 12-6% in the streptokinase group and 13-7% among controls. There was no significant difference in the peak levels or pattern of enzyme increase. The incidence of cardiac failure and reinfarction was similar in the two groups, but major arrhythmias were less common in those on streptokinase (P less than 0-05). In the streptokinase group there were 36 minor and six more serious haemorrhagic complications. Gastrointestinal haemorrhage may have contributed to the death of one patient in each group. There were 18 thromboembolic complications in the streptokinase group and 38 among the controls. Pathological examination of the hearts of 25 patients who had taken streptokinase and 24 controls showed no striking differences between the groups, but haemorrhagic infarcts were found in three patients who had received streptokinase. An infusion of streptokinase within 24 hours of the onset of acute myocardial infarction does not significantly affect the mortality or course of the illness up to six months.


Preoperative skin preparation: clinical evaluation of depilatory cream.

Preoperative hair removal by a depilatory cream was compared with routine shaving. Although the incidence of wound infection was similar in both groups, cream depilation was found to be better. It was effective, atraumatic, non-toxic, and could be self-administered. Furthermore, it could be used safely on granulating wounds and did not support bacterial growth. Depilation was associated with a significant reduction in skin surface bacteria and proved to be cheaper than shaving.


Chemoimmunotherapy of advanced breast cancer: prolongation of remission and survival with BCG.

Forty-five patients with disseminated breast cancer were given a trial of combination chemotherapy consisting of fluorouracil, adriamycin, and cyclophosphamide (FAC) and immunotherapy with BCG given by scarification. The results were compared with those in a comparable group of 44 patients treated with FAC alone immediately before the chemoimmunotherapy study. The remission rates (73% and 76% for FAC and FAC-BCG respectively) were similar in both studies. The durations of remission for patients on FAC-BCG (medium 12 months) were longer than remissions achieved for patients given FAC alone (median 8 months) (P = 0.068). The most notable effect of BCG was on survival. Thus 21 out of 34 patients achieving remission on FAC-BCG were alive at the time of the last follow-up examination (median over 22 months) compared with 11 out of 32 patients achieving remission on FAC (median 15 months) (P = 0.01). Twenty-six of the 45 patients given FAC-BCG were alive at the time of the last follow-up examination (median over 22 months) compared with 12 of the 44 patients given FAC (median 15 months) (P = 0.005). Although the apparent benefit of BCG could be explained by a maldistribution of some prognostic factors, the data suggest that further trial of chemoimmunotherapy of breast cancer should be carried out.


Reduced respiratory responses to carbon dioxide after propranolol: a central action.

In a double-blind study in six subjects propranolol significantly reduced the respiratory sensitivity to carbon dioxide rebreathing. This effect seems to have been due to beta-adrenergic blockade, since it was not seen with D-propranolol. In two subjects increasing doses of propranolol caused progressive reductions in respiratory sensitivity to values below normal and similar to those of patients with ventilatory failure. These changes are probably due to a central action of propranolol.


A new look at the serology of treponemal disease.

The serology of treponemal disease has become simpler and more rational in recent years, mainly as a result of the widespread adoption of specific antibody tests and the use of monospecific fluorescent antibody procedures which give information about the immunoglobulin class of antibodies. A set of tests which has proved particularly useful in routine diagnosis is the following: quantitative TPHA test, quantitative VDRL test, and monospecific (IgG and IgM) FTA-ABS tests. This combination is especially valuable in the assessment of new patients with positive results to serological tests and in the management of patients with treated syphilis.


Immunity in syphilis. Studies in active immunity.

Support for the concept of the development of immunity during the course of syphilis is avaiable in the literature. In experimental syphilis in rabbits, some immunity is present approximately 3 weeks after infection with Treponema pallidum. Resistance to re-infection increases to a maximum at approximately 3 months after infection. Termination of this state by penicillin treatment within this 3-month period may enable re-infection to be accomplished. Attempts to reproduce this state of immunity experimentally by injection of T. pallidum itself, or protein derivatives, or ultrasonic disintegrates obtained from T. pallidum or non-pathogenic treponemes, have been unsuccessful. However, promising results in rabbits have resulted from injecting T. pallidum suspensions attenuated by storage at 4 degrees C, penicillin, or gamma irradiation, and also by suspensions preserved by glutaraldehyde. In the present study, partial resistance to intratesticular challenge in rabbits with T. pallidum has been obtained by immunization with a variety of non-pathogenic treponemes, as exemplified by the strains Nichols, Kazan 2, 4, 5, and 8, Treponema minutum, Treponema ambigua, Treponema refringens and Treponema microdentium. Success is attributed to the processing of immunizing antigens at 4 degrees C and storage until use at -20 degrees C. Attempts to attenuate T. pallidum by immunological means, namely, passage through a limited number of immunized rabbits, were unsuccessful.


A blind controlled trial of dextran 40 in the treatment of ischaemic stroke.

A blind controlled trial of dextran 40 in ischaemic stroke is reported. In the treated group mortality in the acute stage in patients with severe strokes was significantly reduced but survivors were severely disabled and six months later no significant benefit could be detected. In less severe strokes no effect of treatment was found. It is concluded that the action of dextran 40 in reducing acute mortality probably lay in preventing or reversing cerebral swelling but no evidence of the expected action in reducing the area of infarction following improved blood flow was found. The importance of extending assessment beyond the two weeks customary in therapeutic trials in stroke is emphasized.


Somatic sensory cortex of hyrax (Procavia).

Somatic sensory (SI) cortex was mapped in hyraxes by micromapping methods developed and used earlier in slow lorises and llamas. The somatotopic pattern of organization was similar in its general features to that found in other mammals. The perioral and intraoral representations are relatively large in SI of hyrax and exhibit considerable distortion. Sulci are reliable landmarks demarcating boundaries of SI in hyrax as they are in other mammals.


The effect of grinding of human dentine examined by X-ray diffraction, infrared spectroscopy, and electron microscopy.

Analysis of human dentine by infrared spectrophotometry suggests that ball-grinding may result in damage of the apatite crystallites. The present study includes further assessments of this effect by X-ray diffraction, infrared spectroscopy, and electron microscopy. Each of three coarse-ground dentine samples (Group I) was divided into three portions of 30 mg. One of these portions was ball-ground for approximately 1 min (Group II), the second portion for 6 min (Group III), and the third portion for 23 min (Group IV). The 002 reflection showed line broadening, most marked from Group II to III. Electron microscopy showed a gradual change in crystallite appearance with increased grinding, most pronounced from Group II to III. These observations indicate that by prolonged grinding a limit is approached where no further changes in the crystallites occur. Electron microscopy also indicated that fracture of the crystallites might have occurred. This was probably accompanied by strains in the lattice. The infrared spectra indicated that no breakdown of the apatite structure had occurred during the entire grinding.


Physostigmine as an adjuvent to neuroleptanaesthesia in neurosurgical procedures.

The use of physostigmine electively to reverse the effects of droperidal and diazepam has permitted an optimum level of neuroleptanaesthesia in neurosurgical operations where co-operation of the patient is required during part of the procedure. The patient can be put to sleep or readily awakened to be fully co-operative depending on the needs of the surgeon. In this series of seven anaesthetics there were no side effects from the small doses of physostigmine employed. Bradycardia and salivation were not a problem. One child vomited once. Atropine was not necessary. Since a narcotic antagonist is not needed, a reasonable degree of analgesia can be maintained in these patients while they are awake. The latent time for the effect of physostigmine was two to four minutes and the effect of an intravenous dose lasted from 35 to 45 minutes. With physostigmine, these patients wake up gently as though from normal sleep. If neurological assessment is required post-operatively, drowsiness due to drugs can be reversed by giving more physostigmine and the level of consciousness can then be assessed.


Anaesthesia time/dose curves IX: the use of hydromorphone in surgical anaesthesia and postoperative pain relief in comparison to morphine.

Seventy unselected subjects were anaesthetized for major surgical operations with intravenous hydromorphone, nitrous oxide and muscle relaxants as required. The results were compared with earlier observations made with morphine under similar conditions on 44 other subjects from the same hospital population. Hydromorphone was found to be 8.5 times as potent as morphine in terms of the median surgical loading dose, and its median therapeutic half-life was 4.07 hours against morphine's 5.28 hours. It proved to be superior to morphine in its greater consistency of action. The mid-90 per cent variance of its loading dose was 30% less and the same variance of its therapeutic half-life was 65% less than that of morphine. All the differences were statistically significant or highly significant. Observations concerning the fraction of the surgical loading dose of hydromorphone required by the patients to prevent post-operative discomfort also revealed great consistency. This is taken to indicate that psychological factors play only a subordinate role in modulating the intensity of postoperative pain.


Amino acid sequence of penicillopepsin. II. Isolation and characterization of thermolytic peptides.

The determination of the amino acid sequences of 70 peptides obtained from a thermolytic digest of penicillopepsin (EC 3.4.23.7) is described. Fifty-six unique sequences ranging from 2 to 13 amino acids were compiled. Among these was a heptapeptide whose sequence is nearly identical with that of the epoxide-reactive active site peptide of porcine pepsin (EC 3.4.23.1). Considering unrecognized overlaps, a minimum of 272 and a maximum of 293 unique amino acids have been obtained. They account for about 90% of the amino acids of the enzyme.


Application of step-wise discriminant analysis and Bayesian classification procedure in determining prognosis of acute myocardial infarction.

A retrospective study was carried out to assess the feasibility of computer-assisted prognostication by discriminant analysis and the Bayesian classification procedure based on clinical information collected on patients with acute myocardial infarction. The overall accuracy was 94.2% in predicting hospital death but the prediction of late death after discharge was less accurate. It was found that not all of the 44 variables used for analysis were necessary to reach the same level of predictive accuracy--16 to 20 variables would result in almost the identical prediction. The Bayesian classification procedure was applied to estimate probabilities of individual patients belonging to the different prognostic categories.


Long-term therapy of essential tremor with propranolol.

In a double-blind crossover study 12 patients with essential tremor were treated with propranolol and a placebo; 8 improved with propranolol and 3 with the placebo; the degree of improvement with propranolol was greater. In a similar study with diazepam 5 of 12 improved with diazepam and 4 of 12 with the placebo; the degree of improvement was less than that achieved with propranolol. Response in 21 patients to treatment with propranolol for 2 to 4 years was excellent in 4, good in 4 and fair in 10; the condition of 1 was unchanged and that of 2, worse. Excellent response was maintained for as long as 4 years, but response tended to deteriorate with time if initially it was less than excellent. Response decreased with increasing age. No patient 60 years of age or older had an excellent response, and the four with an excellent response were under age 55, three being under age 35; all four had had their tremor less than 12 years. Patients with essential tremor should be given a 3-month trial of propranolol at 120 mg/d; if no significant response is seen the dose should be decreased, then the drug discontinued.


Adjuvant chemotherapy in the surgical treatment of large bowel cancer.

In two large controlled cooperative trials, 1118 patients undergoing surgical resection for large bowel cancer were randomized to be treated by surgery alone or to receive 5-fluorouracil during postoperative followup. The two studies differed slightly in the selection of patients for entry and in the dose regimens of 5-fluorouracil. Following examination of the removed specimen, "curative" and palliative stratifications based on evidence of residual disease were made, so that the two trials actually consisted of five separate patient groups, two groups of patients who had "curative" resections and the other three smaller groups of patients who had palliative resections with residual disease or tumor at a margin of the resected specimen. In all five groups, survival was slightly better in patients receiving chemotherapy although no single difference is statistically significant. Any suggestion that these slight advantages represent a true therapeutic 5-FU effect must be tempered by previous negative adjuvant studies by this group as well as others. Currently the group is conducting a trial using a combination of methyl-CCNU and 5-fluorouracil and has recently added a nonspecific immunostimulant, methanol extraction residue of BCG (MER), to one subgroup.


Fluorouracil, methyl-CCNU and vincristine in cancer of the colon.

In a randomized study 91 patients with metastatic cancer of the colon were treated with either fluorouracil alone or a combination of fluorouracil, methyl-CCNU, and vincristine. Of those treated with the three-drug combination, 37% responded, and of those treated with fluorouracil alone, 22% responded. Median response duration for patients treated with the combination method was 5.0 months and for those with fluorouracil alone, 3.8 months. Toxic effects are also compared. From this study it is concluded that a three-drug regimen provides a definite advantage.


Primary splenic malignant lymphoma, histiocytic type, with sclerosis: report of a case with long-term survival.

A 61-year-old woman with malignant lymphoma, histiocytic type, with slerosis (sclerosing reticulum cell sarcoma) primary in the spleen is described. She has survived clinically free of malignant lymphoma for 11 years after splenectomy therapy alone. This may be the first case of primary splenic malignant lymphoma, histiocytic type, with sclerosis, ever reported in the medical literature.


Hydroxyldaunomycin (Adriamycin) combination chemotherapy in malignant lymphoma.

Combination chemotherapy with CHOP (cyclophosphamide, Adriamycin, vincristine, and prednisone) and HOP (Adrimycin, vincristine, and prednisone, was used as treatment for patients with pathologically staged, advanced non-Hodgkin's lymphoma. Among 204 evaluable patients treated on CHOP there were 71% complete remissions with 92% overall responses. Among the 216 evaluable patients on HOP there were 61% complete remissions and 88% responses. Complete remission rates among patients with histiocytic lymphoma were comparable to those of patients with lymphocytic disease. Patients with nodular lymphoma had higher rates of complete remission than their counterparts with diffuse lymphoma. This was noted with both CHOP (78% vs. 67%) and HOP (67% vs. 60%) induction therapy. Rapid responses were common, as more than 14% of complete remissions and 66% of overall responses were achieved with the first course of treatment. Patients in complete remission have been maintained with either cyclophosphamide, vincristine, and prednisone (COP) or arabinosyl cytosine, vincristine, and prednisone (OAP). After 1 year, 86% of patients on COP and 80% on OAP are projected to be free of disease.


Observations on occult malignant lymphomas in the skin.

Six patients with malignant cutaneous lymphomas are described to show the diversified clinical patterning that may occur. None of the lesions, which ranged from solid nodules, through inflammatory infiltrative sesions, to gangrene, could be identified as malignant lymphoma at the time these patients sought medical attention. Histologic diagnosis of the lymphoma came much later. Pain was a particularly prominent symptom in three of the six patients. The clinician should discard the dictum that malignant lymphoma is a painless process and should not neglect the consideration of malignant lymphoma because of the presence of pain. The skin lesions described provide a model for recognition of extranodal lymphomas elsewhere in the body. Lymphoma should be considered in the presence of unexplained chronic inflammatory processes that affect any organ containing lymphoreticular or epithelial structures.


Comparison of anaphylactic reactions to asparaginase derived from Escherichia coli and from Erwinia cultures.

A retrospective study was undertaken comparing the frequency and severity of anaphylactic reactions to E. coli-derived and Erwinia-derived asparaginase given intravenously on a weekly dosage schedule. Both drugs were found to produce life-threatening hypersensivity reactions with the chance of reaction per dose administered being almost identical--8% for each dose administered. Eleven of 31 patients (35%) experienced anaphylactic reactions, 9/27 (33%) with E. coli and 3/10 (30%) with Erwinia asparaginase, with one patient suffering anaphylaxis to both preparations. A marked increase in the percentage of patients having reactions occurred after the fourth dose of either preparation, with the incidence per dose increasing from 3.3% with the first dose to 32% on the fifth and subsequent doses. Rationale for an antibody-mediated allergic reaction is presented to explain the data.


Cyclophosphamide, vincristine and procarbazine in the treatment of malignant melanoma.

Patients with advanced malignant melanoma were randomly assigned to treatment with cylcophosphamide and vincristine (CV) or cyclophosphamide, vincristine, and procarbazine (CVP). The objective response rates to each regimen (CV 30%, CVP 33%) were the same. Patients with disease confined to skin, subcutaneous tissue nodes, lung, or pleura had a significantly superior response rate to that in those with disease involving liver, bone or brain. The survival of responders was superior to that of nonresponders. The efficacy and moderate toxicity of CV suggests that further work with this combination in malignant melanoma is indicated.


Randomized prospective trial of DTIC (NSC-45388) alone versus BCNU (NSC-409962) plus vincristine (NSC-67574) in the treatment of metastatic malignant melanoma.

Fifty patients with metastatic malignant melanoma were randomized to treatment with either DTIC (2 mg/kg/day X 10 iv) or the combination of BCNU (150 mg/m2 iv) plus vincristine (VCR) (2 mg/m2 iv on Day 1 only). Treatment failures were crossed over to the alternate therapy. Primary, secondary, and cumulative response rates to DTIC were 29%, 9%, and 22%, respectively. Primary, secondary, and cumulative response rates to BCNU plus VCR were 23%, 29%, and 25%, respectively. Five of 26 patients (19%) experienced objective regression from secondary therapy after failure to respond to primary therapy. DTIC produced gastrointestinal and hematologic toxic effects; BCNU plus VCR produced gastrointestinal, hematologic, and neurologic toxic effects. VCR administered at a dose of 2 mg/m2 resulted in excessive neurologic toxic effects in 12 of 21 patients; a maximum VCR dose of 2 mg/injection was well tolerated by 15 subsequent patients without an adverse effect upon response rate. An analysis of tumor burden and organ involvement in responders and nonresponders suggests that DTIC is the first-choice treatment for patients with limited tumor burdens and nonvisceral metastases; BCNU plus VCR is the first-choice treatment for patients with extensive tumor burdens and visceral-predominant disease. However, failure to respond to primary therapy does not preclude response to secondary therapy with the alternate regimen.


Bioassay of plant extracts for anticancer activity.

Crude plant products were first tested in the Drug Research and Development Program, Division of Cancer Treatment, NCI, in 1956. The first materials were voluntarily submitted and later materials were acquired through various contracts. A review is given of the tumor systems which have been used for the evaluation of these plant products and of in vitro assays utilized to expedite fractionation and isolation.


Types of anticancer agents isolated from plants.

The active compounds which have been isolated from plants and tested in the chemotherapy program of the National Cancer Institute since the inception of the plant program (as part of the Cancer Chemotherapy National Service Center) are listed, classified into types, and discussed in terms of their activity in experimental tumor systems. The tumor systems include the most important ones comprising the regular screen at different times and also the slow-growing tumors, B16 melanoma and Lewis lung carcinoma (new). The structure-antitumor activity relationships bring out the desirability for further investigation of certain types of compounds as possibilities for clinical trial. Notes on the current pharmacologic anc clinical status of certain compounds are also presented.


Distribution of anticancer activity in higher plants.

The systematic distribution in higher plants of general anticancer activity, high-interest anticancer activity, and high-interest compounds is presented. Special emphasis is placed on those taxonomic groups offering, by virtue of their high-interest activity and/or compounds, the greatest potential as sources of useful anticancer agents.


Novel plant-derived tumor inhibitors and their mechanisms of action.

The activity-directed isolation of new tumor inhibitors of plant origin has yielded many novel compounds with significant growth-inhibitory properties. A large proportion of the new compounds contain highly electrophilic functionalities and chemical and biochemical studies are yielding a growing body of evidence to support the view that these compounds may act by selective alkylation of growth-regulatory biologic macromolecules. The selectivity may result from many factors, among which are transport of the tumor inhibitor into the cell and the chemical nature and steric environment of the specific nucleophile to be alkylated. Model studies support the hypothesis that the inhibition of tumor growth by the new agents may be attributable to selective alkylation of key enzymes which control cell division.


The genus Cephalotaxus: source of homoharringtonine and related anticancer alkaloids.

Various phases of research on Ciphalotaxus alkaloids are summarized, including their discovery, nature of antitumor activity, isolation, structural characterization, synthesis, biogenesis, and mechanism of physiologic action.


Oncogenic and tumor-promoting Spermatophytes and Pteridophytes and their active principles.

A survey and discussion are presented of plants classified as Spermatophyta and Pteridophyta, extracts of which have been shown to be oncogenic or tumor-promoting in animals. The active oncogenic and tumor-promoting principles, where known, have been identified. They represent tannins; pyrrolizidine, indole, tropolone, quinoline, purine, and benzophenanthridine alkaloids; nitroso compounds; triterpene glycosides; lignans; isoflavans; allyl benzenoids; simple (nu-pyrenes; and carbocyclic hydroxy acids. A total of 28 compounds of known structure have been identified as oncogens and several phorbol esters as tumor-promoters. Plants known to contain any of the 28 oncogens (excluding shikimic acid and caffeine) have been tabulated; they represent at least 454 species, 110 genera, and 34 families of Spermatophyta and Pteridophyta.


Plant folklore: a tool for predicting sources of antitumor activity?

The National Cancer Institute's record of "active plants" (extracts which showed a significant inhibitory effect in experimental tumor systems) was compared with plants reported in folklore to have medicinal or poisonous properties. The occurrence of active plants was found to be higher in plants reported in folk literature than in plants collected at random, suggesting a correlation between plants used in folklore and those with anticancer activity.


Immediate hypotensive after-effects of posterior hypothalamic lesions in awake rats with spontaneous, renal, or Doca hypertension.

If the posterior hypothalamus contributes to elevate blood pressure in hypertension by increasing sympathetic vasomotor activity, then lesions of the posterior hypothalamus should lower blood pressure more in hypertensive than in normotensive rats. To test this hypothesis without complications caused by anaesthesia, aortic pressures were recorded from indwelling catheters in awake rats before and after selective hypothalamic destruction. In normotensive rats rats, bilateral lesions of the medial areas of the posterior hypothalamus always lowered blood pressure while those in the anterior hypothalamus slightly increased it. Heart rate responses varied widely and did not seem to contribute to the blood pressure changes. Posterior hypothalamic lesions of approximately the same size had significantly greater hypotensive after-effects in renal and spontaneously hypentensive rats than in normotensive or Doca hypentensive ones. These results imply that sympathetic overactivity emanating from posterior hypothalamic centres contributes to the blood pressure elevation in spontaneous or chronic renal hypentension but not in Doca hypertension. However, because of inherent weaknesses in the 'lesion method' and the complexity of blood pressure regulation in awake animals, other explanations are possible.