Effect on survival after myocardial infarction of long-term treatment with phenytoin.

A prospective, randomised, open trial was performed in 150 patients to test for any beneficial effects on 2-year mortality of long-term antiarrhythmic therapy with phenytoin in patients with acute myocardial infarction. Patients were stratified according to age, sex, past history of myocardial infarction, and the presence of absence of electrical or mechanical complications in the course of acute infarction. They were then randomised to treatment or control groups (74 v. 76). The former received phenytoin in doses aimed at maintaining plasma phenytoin levels between 40 and 80 mumol/litre. All patients entered the study before discharge from the coronary care ward. Plasma phenytoin levels were in the therapeutic range in between 51 and 75 per cent of subjects at any follow up visit. There were 19 withdrawals from the treatment group, 10 of which were the result of side effects. There were 5 withdrawals from the control group. According to the original intention to treat, there were 18 deaths at 2 years in the treatment group and 14 deaths in the control group. There was no reduction in the incidence of instantaneous or sudden deaths. Deaths on treatment were not associated with a low phenytoin plasma level. Phenytoin treatment showed no beneficial effects on mortality and was associated with a high incidence of side effects.


Methods for assessment of the effect of drugs on cerebral blood flow in man.

The cerebral circulation is not an unimportant vascular bed but it is difficult one to investigate. Methodological difficulties are the main reasons for the paucity of clinical pharmacology studies to date. Of the methods currently available those of the McHenry (1964) and Wyper et al. (1976) appear to be the most useful to the clinical investigator. Increasing use of multiple detectors even with inhalation techniques may give evidence of regional drug effects upon the cerebral circulation. Improvements in methodology in the last few years, particularly the development of atraumatic methods are likely to act as an important spur to research in this field.


A controlled trial of labetalol (Trandate), propranolol and placebo in the management of mild to moderate hypertension.

1. Labetalol, a new drug combining alpha-and beta-adrenoceptor blocking properties, has been compared with placebo in a double-blind crossover study of a group of patients with mild to moderate essential hypertension (blood pressure 150/100 to 189/114 mmHg). 2. Labetalol and propranolol lowered blood pressure satisfactorily in the supine position, but labetalol reduced blood pressure more in the erect posture and following exercise and induced less bradycardia. Thus alpha- as well as beta-adrenoceptor blocking actions appear to contribute to blood pressure reduction. 3. Side effects attributable to labetalol were few. The effective dose ratio labetalol: propranolol was 2.5:1 (w/w). 4. Labetalol, a new form of hypotensive agent, merits further controlled assessment of  its usefulness in relation to existing drugs.


Responses of human and baboon arteries to prostaglandin endoperoxides and biologically generated and synthetic prostacyclin: their relevance to cerebral arterial spasm in man.

1 Isolated strips of human or baboon basilar, middle cerebral, vertebral or common carotid arteries were set up in an isolated organ bath or in a superfusion cascade system. 2 These arteries relaxed to prostacyclin but contracted to prostaglandin endoperoxide (PGH2). 3 Human and baboon isolated arteries also generated prostacyclin from exogenous endoperoxide (PGH2). 4 Human arteries generated prostacyclin 36 h post-mortem but not 40 h post-mortem. The biologically generated prostacyclin relaxed the basilar artery and overcame the contractile effects of PGH2. 5 Thromboxane A2-like activity generated during human platelet aggregation by arachidonic acid caused contractions of the human basilar artery. 6 Prostacyclin reversed contractions of human basilar arteries caused by an unidentified vasoconstrictor factor in cerebrospinal fluid obtained from patients with cerebral arterial vasospasm after subarachnoid haemorrhage following rupture of cerebral arterial aneurysms. 7. The above vasospasm may be due at least in part to disordered physiological control of the calibre of cerebral arteries caused by diminished synthesis of prostacyclin.


Cinnarizine in the treatment of chronic asthma.

1 Cinnarizine, an inhibitor of calcium ion transport across smooth muscle cell membrane, has been shown to exert an anti-asthmatic effect in patients with chronic asthma. 2 It is postulated that antagonism to calcium ion transport across the mast cell membrane may cause the compound to have a pharmacological effect similar to sodium cromoglycate. 3 Cinnarizine is orally active and its therapeutic effect is demonstrated in a double-blind, cross-over, placebo controlled study. 4 Patient benefit was shown by a significant improvement in peak flow rate. A non-significant trend towards a reduction in symptomatic bronchodilator usage and a decrease in asthma symptom score was also shown. 5 It is concluded that cinnarizine could well prove to be the first of a new family of anti-asthmatic drugs offering a protective effect when taken systemically.


Development of a model for transepidermal elimination.

Transepidermal elimination is a phenomenon which occurs spontaneously in certain skin disorders in which effete connective tissue or foreign materials is expelled via the epidermis to the exterior. In order to learn how this phenomenon is effected and what factors control it, we have attempted to produce an animal model for the process. When charcoal particles are deposited subepidermally in guinea-pig flank skin transepidermal elimination occurs within 4 days and we believe that this mimics the disorder seen in man. The hair follicles became hyperplastic and were intimately involved in the reaction, some of the particles actually being expelled via the follicular lumina. The reaction has some similarities to the wound healing response.


Absence of cutaneous IgA in coeliac disease without dermatitis herpetiformis.

Skin biopsies from 17 patients with coeliac disease, but without dermatitis herpetiformis, were examined by direct immunofluorescence. In none of them was IgA detectable. In contrast, IgA was present in dermal papillae and/or in relation to the dermo-epidermal junction in 80 out of 83 patients with dermatitis herpetiformis. These findings confirm that cutaneous deposition of IgA at these sites is a very characteristic feature of dermatitis herpetiformis and is unrelated to coeliac disease itself.


Ultrastructural demonstration of T cells in cutaneous tissue sections using specific anti-human T cell antiserum.

The presence of T lymphocytes in the situ in cutaneous infiltrates of contact dermatitis and lichen planus has been demonstrated by an immunoelectronmicroscope technique. A specific anti-human T cell antiserum labelled with peroxidase-conjugated anti-immunoglobulin serum was used to reveal the specific human T lymphocyte antigen (HTLA) which was visualized ultrastructurally. This technique shows that labelling is limited to the surface membrane and allows study of the in situ interrelationship of T cells.


Bullous pemphigoid induced by PUVA therapy. Is this the aetiology of the acral bullae produced during PUVA treatment?

Among 24 psoriatic patients treated with PUVA therapy, five developed bullae. Direct and indirect immunofluorescence studies supported the diagnosis of bullous pemphigoid in one patient. In the others, the blisters were a phototoxic acral eruption.


Industrial bronchitis.

For many years there has been much argument whether workers in the dusty trades are prone to chronic bronchitis. In 1966 the Medical Research Council issued a report of a Select Committee which concluded that occupationally induced bronchitis did not play a significant part in the aetiology of airways obstruction in dust-exposed men. Since then epidemiological studies have demonstrated that the prolonged inhalation of dust leads to an increase in prevalence of cough and sputum. Furthermore, new physiological techniques have demonstrated a slight decrement in ventilatory capacity as a result of industrial bronchitis, and which is related to lifetime dust exposure. Unlike bronchitis induced by cigarette smoke, the predominant effect of industrial bronchitis is on large rather than small airways and the condition is not accompanied by emphysema.


Neonatal head circumference and the treatment of maternal hypertension.

In a random controlled trial of methyldopa for the treatment of hypertension in pregnancy presenting before 28 weeks gestation, the newborn in the treated group had relatively smaller head circumferences. This difference persisted at two months of age when correction had been made for birth weight, gestation and sex, but was no longeer detectable at six or twelve months. Within the treated group no relationship was found between neonatal head circumference and the total amount or duration of methyldopa received during pregnancy. Comparison of treated and untreated groups according to the time of entry to the study showed that significant differences in neonatal head circumference were only present in patients who entered between 16 and 20 weeks gestation. It is possible that this could be a sensitive period for the interaction of fetal head growth and the onset of specific treatment in hypertensive pregnancy.


A comparison of four methods of ripening the unfavourable cervix.

A comparison was made between four methods of ripening the unfavourable cervix (extra-amniotic prostaglandin E2 gel, oral prostaglandin E2, intravaginal prostaglandin E2 and intravenous oxytocin) in a clinical trial involving 60 primigravidae. In all groups there was an improvement in cervical status. This was significantly greater in those patients who received extra-amniotic prostaglandin gel and they also showed significant decreases in the mean induction-delivery interval and in the incidence of Caesarean section.


Serum prolactin and gonadotrophin levels before and after luteinizing hormone-releasing hormone in the investigation of amenorrhoea.

An intravenous luteinizing hormone-releasing hormone (LRH) test was performed in 287 women with amenorrhoea. Prolactin, progesterone and oestrogens in serum were also measured. Twenty-four women with premature ovarian failure and 9 with gonadal dysgenesis had raised basal follicle-stimulating hormone (FSH) levels. Neither the basal luteinizing hormone (LH) level nor the gonadotrophin responses after LRH gave a better separation of this group of women with irreversible ovarian failure. Measurement of prolactin levels were valuable in that 15 of 42 patients with hyperprolactinaemia had a radiologically abnormal pituitary fossa, whereas pituitary fossa abnormalities were found in only 11 of 245 normoprolactinaemic women. It was thought that 181 women had functional amenorrhoea; 54 per cent of these women had developed amenorrhoea in relation to weight loss and 32 per cent in relation to discontinuation of oral contraceptives. A strong correlation was found between the body weight and the basal gonadotrophin levels. The basal LH levels were correlated with serum oestrogen levels, the basal FSH level and the LH response to LRH. Most of the patients with low basal LH values had developed amenorrhoea in relation to self-imposed weight-loss. The responses to LRH were often impaired in the underweight patients but became normal after weight gain. The polycystic ovary syndrome (PCO) could not be diagnosed by measuring either basal or LRH-stimulated gonatrophin levels. Single FSH and prolactin determinations in serum seemed to be the only indispensible hormone assays in the routine clinical evaluation of amenorrhoea.


The effect of cervical encerclage on plasma prostaglandin concentrations during early human pregnancy.

In women having either cervical encerclage under general anaesthesia or a vaginal examination in the early second trimester of pregnancy, peripheral plasma levels of 13,14-dihydro-15-keto prostaglandin F (PGFM) were measured before and after each procedure. A significant rise in circulating levels of PGFM was found within minutes of completing cervical encerclage but not after vaginal examination or induction of general anaesthesia.


Microfilaments in the cells of the human trabecular meshwork.

In this present study the results are presented of a combined ultrastructural and immunofluorescent investigation of 'smooth muscle' elements within the cytoplasm of human trabecular meshwork cells. The cytoplasm of human meshwork cells both in vivo and in vitro is replete with 10 nm intermediate filaments and also contains smaller 6 nm filaments which are particularly prominent in the cell processes. By immunofluorescence using sera rich in antibodies to contractile proteins, particularly actin, cultured meshwork cells showed strong cytoplasmic fluorescence. On occasion the cytoplasmic fluorescence was diffuse, but more often recognisable bundless (stress fibres) or a loose fibrillar framework was found. The possible role of structural and contractile cellular proteins in trabecular function was discussed.


Piloplex, a new long-acting pilocarpine polymer salt. B: Comparative study of the visual effects of pilocarpine and Piloplex eye drops.

Induced accommodation and changes in vision (distance and near) were measured monocularly and binocularly in 9 young healthy volunteers in a double blind study after administering to them pilocarpine hydrochloride 4%, Piloplex 3.4, and saline eye drop instillations. Piloplex 3.4, a new long-acting pilocarpine polymer salt, and pilocarpine hydrochloride 4% (both contain equal amounts of pilocarpine-3.4%) induced changes in vision and accommodation. These changes were greater with pilocarpine hydrochloride than with Piloplex. The maximum changes occurred half an hour after instillation and the effect vanished after an additional period up to 3 hours. The changes were greater when measured monocularly than binocularly. Piloplex initiates a prolonged hypotensive effect which lasts for 12 hours. Patients with glaucoma are thus able to use Piloplex on a twice-daily schedule. Consequently, visual disturbances occur only once a day in contrast to pilocarpine hydrochloride given 4 times a day, which induces 3 visual disturbances during the day.


Evaluation of a 5% guanethidine and 0.5% adrenaline mixture (Ganda 5.05) and of a 3% guanethidine and 0.5% adrenaline mixture (Ganda 3.05) in the treatment of open-angle glaucoma.

A trial of a mixture of guanethidine 5% and adrenaline 0.5% (Ganda 5.05) and of guanethidine 3% and adrenaline 0.5% (Ganda 3.05) was conducted on 90 eyes in 53 patients with open-angle glaucoma or ocular hypertension. The cases fell into 5 groups: untreated cases, cases on pilocarpine 1%, on pilocarpine 2%, on pilocarpine 2 to 4% and adrenaline 1%, and on separate guanethidine 5% and adrenaline 1%. Baseline pressures and average pressures on the previous treatment were established. Substitution with Ganda 3.05 or 5.05 was started, and the patients attended 2 weeks, 1 month, 3 months, and 6 months from the start of the trial. Applanation tonometry was carried out at the same time of day. The pupil was measured, ptosis and superficial punctate corneal staining were looked for and evaluated, and the patients were questioned for symptoms of side effects and acceptability. All the eyes that had previously been treated with pilocarpine 1% or 2% presented significantly lower intraocular pressures on Ganda 3.05. The patients on pilocarpine 4% and adrenaline 1% also had lower intraocular pressures on Ganda 5.05, but the significance was less, and the patients on separate guanethidine and adrenaline had a small but not statistically significant drop in pressure. Ptosis and discomfort were evaluated on a subjective scale. Patient acceptability was good. The trial was interrupted in 5 cases for various reasons. Tachyphylaxis and tolerance to the mixtures were not observed in this series.


Ocular and cardiovascular effects of local and systemic pindolol.

Ocular and cardiovascular effects of topical and intravenous pindolol have been studied in a balanced cross-0ver double-blind trial in 6 healthy volunteers. When applied to 1 eye pindolol lowered intraocular pressure in both the treated and untreated eyes with only minimal reduction in resting pupil diameter and light reflex response. The concentration in plasma was much lower and inhibiton of exercise tachycardia about half that when the same dose was administered intravenously. The findings suggest that beta-adrenoceptor blocking drugs should not be used in the treatment of glaucoma in patients who also suffer from heart failure.


Characteristics of proline transport in normal and starved cells of Candida albicans.

1. There was no apparent correlation between the rate of respiration and rate of accumulation of proline in Candida albicans cells. 2. In contrast to normal cells, the respiration in the starved cells became completely cyanide insensitive. The starvation of cells in the presence of cycloheximide prevented the cells from becoming cyanide insensitive. The addition of Fe(III), however, accelerated the process. 3. Oxidizable substrates e.g. NADH, acetate and glucose, when added to cyanide-insensitive starved cells, exhibited 40--280% stimulation in respiration rate. However, this enhancement in oxidation by various substrates was not coupled to a simultaneous increase in the proline uptake or in intracellular ATP levels. 4. There was 6-fold stimulation in proline uptake when cyanide-insensitive cells were preincubated with 50 mM glucose. The preincubation of starved cells resulted in a partial restoration of cyanide sensitivity and increased intracellular ATP levels. The preincubation of starved cells with other oxidizable substrates resulted in a partial restoration of cyanide sensitivity but had no stimulatory effect on intracellular ATP levels and proline accumulation. 5. Both the enhanced uptake and ATP levels in glucose preincubated cells were found to be completely abolished by iodoacetate. 6. It is proposed that the increased proline uptake in cells preincubated with glucose was mainly due to the production of glycolytic energy.


Studies on the mode of action of hygromycin B, an inhibitor of translocation in eukaryotes.

Hygromycin B is an unusual aminoglycoside antibiotic active against both prokaryotic and eukaryotic cells. Hygromycin B at 0.38 mM concentration completely halts yeast cell growth in rich media, presumably by preventing protein synthesis by cytoplasmic ribosomes. Polypeptide synthesis in cell-free extracts from rabbit reticulocytes, wheat germ and yeast is strongly blocked by low concentrations of hygromycin B. The antibiotic inhibits peptide chain elongation by yeast polysomes by preventing elongation factor EF-2-dependent translocation, although it does not affect either the formation of the EF-2-GTP-ribosome complex or the EF-2- and ribosome-dependent GTP hydrolysis which takes place uncoupled from translocation. The inhibition of translocation by hygromycin B might result from the stabilization of peptidyl-tRNA bound to the ribosomal acceptor site, since the stability of [3H]Phe-tRNA-EF-1-poly(U)-ribosome and [3H]Phe-tRNA-poly(U)-ribosome complexes is increased in the presence of hygromycin B. The inhibition of polyphenylalanine synthesis by reticulocyte ribosomes and enzymic translocation of peptidyl-tRNA by yeast polysomes can be reversed by increasing concentrations of EF-2 suggesting a relationship between the binding sites of EF-2 and hygromycin B on the ribosome. Neither non-enzymic translocation, that takes place in the presence of high potassium concentrations, nor the peptide bondforming step are affected by hygromycin B.


Cross-linking and relaxation of supercoiled DNA by psoralen and light.

Photoreaction of 4,5',8-trimethylpsoralen with superhelical ColE1 and ColE1amp DNA was studied. Changes in mobilities in agarose gels, formation of interstrand cross-links, and DNA strand breaks were determined. Psoralen and light treatment removed negative superhelical turns, and extensive treatments failed to produce positive superhelical turns in covalently closed plasmid DNA. The rate of relaxation of superhelical turns by psoralen Photobinding appeared to be directly proportional to the number of superhelical turns remaining. A unique reaction mechanism is presented to explain these results. By this interpretation the initial rate of psoralen photobinding to superhelical DNA was estimated to be 3 times that for linear DNA, and the ratio of cross-linking to monofuctional adducts appears to be dependent on the superhelical conformation of the DNA. The estimated ratio of psoralen molecules bound to DNA strand breaks was 1.7 . 10(4):1, and 70% of this breakage is caused by the light alone.


Ultracentrifugation studies of yeast valyl-tRNA synthetase and of its interaction with tRNAVal.

Yeast valyl-tRNA synthetase and its complexes with yeast tRNAVal were investigated by means of analytical ultracentrifugation. A molecular weight of 125 700 +/- 1500 and a sedimentation coefficient (SO 20, w) of 6.3 +/- 0.3 were found for the native enzyme. When the enzyme (3--60 muM) was mixed with its cognate tRNA, several types of complex were observed, depending on the relative amounts of the two macromolecules. In the presence of equimolecular amounts of tRNA and enzyme, a complex formed by the association of one of each molecule was observed with a sedimentation coefficient of about 7.3 S. However, for tRNA/enzyme stoichiometries lower than one, beside the 1 : 1 complex, a complex of higher molecular weight was observed, with a sedimentation coefficient of about 10.0 S which fits with the association of two valyl-tRNA synthetase molecules with one tRNA molecule. This 2 : 1 complex was predominant from tRNA/enzyme stoichiometries lower than 0.3. It dissociated into the 1 : 1 complex upon addition of monovalent salts or MgCl2, suggesting the electrostatic nature of the interaction in this association. All these association and dissociation phenomena were detected over a large range of pH (6.0--7.5) and in various buffers.


Isolation and characterisation of 14-S RNA from spinach chloroplasts.

14-S RNA was purified from spinach chloroplasts. It has a molecular weight of 0.43 . 10(6) and the following nucleotide composition: 20% CMP, 23.9% AMP, 24.2% GMP and 31.9% UMP. The accumulation of 14-S RNA in chloroplasts of cotyledons of dark-grown plants is stimulated by light. Conditions are described for the isolation of 14-S RNA in the absence of appreciable fragmentation of chloroplast 23-S rRNA and the evidence that it represents a distinct type of chloroplast RNA is discussed. Translation of 14-S RNA in a protein synthesising system from Escherichia coli gives rise to two polypeptides with molecular weights of 13 200 and 12 600 and the possible role of 14-S RNA as a chloroplast messenger is discussed.


Guanine nucleotide metabolism in a mutant strain of Escherichia coli with a temperature sensitive lesion in rRNA synthesis.

We have described a mutant of Escherichia coli (designated 2S142) which shows specific inhibition of rRNA synthesis at 42 degrees C. ppGpp levels increase at the restrictive temperature, as expected. However, when the cells are returned to 30 degrees C, rRNA synthesis resumes before ppGpp levels have returned to normal. Furthermore, when ppGpp levels are decreased by the addition of tetracycline or choramphenicol, rRNA synthesis does not resume at 42 degrees C. Also, a derivative of 2S142 with a temperature-sensitive G factor (which cannot synthesize either protein or ppGpp at 42 degrees C) shows identical kinetics of rRNA shut-off at 42 degrees C as 2S142. Thus, the elevated ppGpp levels in this mutant do not appear to be directly responsible for the cessation of rRNA synthesis at 42 degrees C.


Modification of rat liver chromatin by N-methyl-N'-nitro-N-nitrosoguanidine or N-ethyl-N'-nitro-N-nitrosoguanidine and template activity for RNA synthesis by Escherichia coli RNA polymerase after reconstitution.

Rat liver chromatin was fractionated into DNA, histones and non-histone chromosomal proteins and each component was modified with N-methyl-l-N'-nitro-N-nitrosoguanidine of N-ethyl-N'-nitrosoguanidine. The radioactivity of 14C-labeled alkyl or guanidino moieties of both compounds bound significantly to both histones and non-histone chromosomal proteins and the binding of N-methyl-N'-nitro-N-nitrosoguanidine was higher than N-ethyl-N'-nitro-N-nitrosoguanidine. However the binding of both compounds to DNA was very low and its significance was hard to evaluate. All of the three components, one of which was modified, were reconstituted into chromatin, then, [3H]UMP incorporation into acid insoluble material using Escherichia coli RNA polymerase (EC 2.7.7.6) was measured. Only with the reconstituted chromatin containing histones modified either by N-methyl-N'-nitro-N-nitrosoguanidine or N-ethyl-N'-nitro-N-nitrosoguanidine, the template activity increased drastically; i.e., about 10 or 5 times higher than that with the unmodified reconstituted chromatin, respectively. However, any remarkable alteration in the electrophoretic pattern of protein fraction of the reconstituted chromatin could not be found. The results obtained in this study are discussed in the context that the modified histones could give rise to change in the mutual interaction of chromosomal components during the reconstitution of chromatin accompanied with the increase of chromatine template activity.


Effect of estrogen on gene expression in the chick oviduct. In vitro transcription of the ovalbumin gene.

Problems involved in using the Hg-nucleotide technique for in vitro chromatin transcription are 2-fold. First, Escherichia coli RNA polymerase can utilize endogenous RNA as template and synthesize complementary sequences which remain base-paired to the template, thereby allowing it to bind to the SH-Sepharose column and copurify with the newly synthesized Hg-RNA. Second, non-mercurated endogenous RNA can bind to the SH-Sepharose through aggregation with Hg-RNA and thus be retained in the final RNA preparation. These two problems associated with the Hg-nucleotide technique can be minimized by modifying the conditions for RNA synthesis and SH-Sepharose chromatography. Using the modified procedure the Hg-nucleotide and SH-Sepharose technique can remove more than 90% of endogenous RNA contaminants. In order to directly demonstrate that the mRNAov sequences detected in vitro result from de novo transcription of oviduct chromatin, experiments were carried out which show that the hybridizable RNA sequences  contain the Hg element and that the synthesis of these RNA sequences is sensitive to low concentrations of actinomycin D. These combined results strongly suggest that the majority of mRNAov sequences detected by hybridization to  cDNAov is indeed due to DNA-dependent RNA synthesis by E. coli RNA polymerase and not due to an artifact of endogenous RNA contamination. This observation was further supported by data obtained using a filter hybridization method which measures directly the mRNAov sequences present in [3H]RNA synthesized from chromatin. The 3H-labeled ovalbumin messenger RNA was assayed by hybridization to cloned pOV230 DNA containing the ovalbumin structural gene sequence. With this modified Hg-nucleotide-SH-Sepharose technique and filter hybridization technique, we have restudied the in vitro transcription of the ovalbumin gene from chromatins isolated at different stages of hormone-induced oviduct development. The results are in agreement with our previous findings which suggest that the primary regulation of ovalbumin synthesis by steroid  hormones occurs at the transcriptional level.


Non-histone chromatin proteins of B lymphocytes stimulated by lipopolysaccharide.

The synthesis of non-histone chromatin proteins and nucleoplasmic proteins has been followed during lipopolysaccharide-induced division and differentiation of murine B lymphocytes. Synthesis was measured by pulse labelling with [3H]leucine, extraction of proteins was under conditions designed to prevent proteolysis and analysis of labelled proteins was by polyacrylamide gel electrophoresis. The average specific activity of non-histone chromatin proteins increased 3-fold, to a maximum, after culture for 24 h with lipopolysaccharide. Comparison of the relative synthesis of individual proteins (stimulation index) reveals three distinct responses: (1) those in the largest group show low stimulation indices, generally less than two; (2) a group of four proteins have indices between 4 and 5; (3) two proteins (molecular weights 21 000 and 22 000) both show an index of 5 at 24 h rising to between 7 and 8 by 48 h when the average specific activity is falling, coinciding with the period of rapid differentiation to high rate IgM secretion. Additionaly at this time, a newly labelled protein (Mr = 36 500) appears in the nucleoplasm followed by a second protein (Mr = 63 000) appearing between 48 and 72 h. The patterns of change are consistent with an overall increase in non-histone chromatin proteins synthesis, necessary for cell division, with superimposed specific changes in synthesis of non-histone chromatin proteins which could be related to regulation of cell differentiation.


Methionyl-tRNA synthetase of Escherichia coli. A zinc metalloprotein.

The native dimeric form of methionyl-tRNA synthetase of Escherichia coli contains two zinc atoms per dimer, one per subunit. The bound zinc is retained upon trypsin modification which yields a monomer with one zinc atom. The enzymatic activity of both the dimeric forms is reversibly inhibited by 1,10-phenanthroline but not by its non-chelating analogues. In addition, the native enzyme binds two Mn2+ per dimer with a binding constant of approx. 70 micron but no binding is observed with the trypsin-modified monomer.


Studies on modification of tryptophan, methionine, tyrosine and arginine residues of human follicle-stimulating hormone and its subunits.

Based on the regeneration of the hormonal activity following recombination, the alpha and beta subunits of human follicle-stimulating hormone have been designated as 'functional' or 'nonfunctional'. Chemical modifications of the tryptophan, methionine, tyrosine and arginine residues of human follicle-stimulating hormone, luteinizing hormone, and the 'functional' human follicle-stimulating hormone alpha and beta subunits have indicated that the tryptophan in human follicle-stimulating hormone-beta and human luteinizing hormone-beta is essential for the biological activity. The iodination of human follicle-stimulating hormone-alpha did not interfere with the hormonal activity. The modification of arginine abolishes the biological activity of the hormones. The accessibility of tyrosine and methionine in human follicle-stimulating hormone-alpha, of arginine in both native hormones and subunits, and the non-availability of the tryptophan residues to 2-hydroxy 5-nitrobenzyl bromide suggest that the alpha subunit lies on the surface of the native molecule.


[Multiplicity of monoamine oxidase: inhibition of mitochondrial monoamine oxidase activity by isopropylhydrazide of D,L-serine].

Isopropylhydrazide of D,L-serine (IHS) inhibits by 50% (at 37 degrees for 10 min) deamination of serotonin or beta-phenylethylamine by monoamine oxidases from bovine brain stem mitochondrial membranes at the 2.6 X X 10(-5) M or 9 X 10(-5) M, respectively. In order to inhibit by 50% the deamination of tyramine under the same conditions a considerably lower (2.5 X X 10(-6) M) concentration of IHS is required. Kinetic studies of inhibition of enzymatic deamination of all the three biogenic monoamines by IHS showed that the irreversible blocking of the monoamine oxidase activity is preceeded by formation of dissociating enzyme-inhibitor complexes. Values of the dissociation constants of these complexes measured (at 37 degrees) with serotonin, phenylethylamine or tyramine as substrates for estimation of the residual monoamine oxidase activity are 0.47; 0.13 or 0.023 mM, respectively. Significant differences are also found between thermodynamic and activation parameters characterizing both  both steps of interaction between IHS and the monoamine oxidases of mitochondrial membranes in the experiments with serotonin, phenylethylamine or tyramine as substrates. The data obtained suggest the existence of different monoamine oxidases (or their active sites) catalyzing oxidative deamination of serotonin, phenylethylamine or tyramine in the fragments of mitochondrial membranes from bovine brain stem.


[Albumin mRNA from rat liver cells. Analysis of immunoadsorption of individual polyribosomes].

The analysis of albumin polyribosomes immunoadsorption is carried out using "sandwich" immunoadsorbents prepared on the basis of two aminobenzylcelluloses: commercial (paraaminobenzylcellulose) and synthesised (methaaminobenzyloxymethylcellulose). A method is worked out which is good for the estimation of the adaptibility of different aminobenzylcellulose preparations as an insoluble basis for the immunoadsorbent. Major properties of the "sandwich" sorbent (the accessible capacity and specificity) and the percent of isolated individual polyribosomes are found to be interrelated and determined by conditions of the immunoadsorption reaction. The increase of polyribosomes and sorbent concentrations and their ratio in the incubation medium results in the increase of the sorbent accessible capacity and the decrease in the inspecific adsorption but at the same time the percent of adsorbed polyribosomes decrease too. The "excess" of adsorbent with respect to polyribosomes, participating in the binding reaction, is necessary for the quantitative isolation of individual polyribosomes.


Agar in control of hyperbilirubinemia of full-term newborn infants with erythrocyte G-6-PD deficiency.

40 full-term newborn infants with erythrocyte glucose-6-phosphate dehydrogenase (G-6-PD) deficiency were used for a study concerning the effectiveness of agar per os in preventing severe hyperbilirubinemia. 20 randomly selected neonates were given agar (1 g/kg/day) orally in 4 daily doses from their 1st to their 5th day of life. 20 infants were not treated and served as controls. Three exchange transfusions were performed in the experimental as well as in the control group. According to these results, agar does not seem to be effective in preventing severe hyperbilirubinemia, which frequently occurs in newborn infants with erythrocyte G-6-PD deficiency.


The protective effect of endogenous interferon in mouse malaria, as demonstrated by the use of anti-interferon globulins.

Both death rate and percentage of parasitized erythrocytes in mice infected with Plasmodium berghei were enhanced by injections of anti-interferon globulins. As, in the same time, parasite-induced interferon was neutralized by these globulins, it can be concluded that endogenous interferon plays an important inhibiting role during parasitic diseases, such as malaria, as it has been previously demonstrated in many virus infections.


Double-strand breaks from single photochemical events in DNA containing 5-bromouracil.

Ultraviolet irradiation of Escherichia coli cells with a low level of 5-bromouracil incorporated produces DNA double-strand breaks by single photochemical events, one such break per 100 single-strand breaks, the latter assayed in alkali-denatured DNA. About 2.5--4 double-strand breaks are produced per "lethal hit," compared with about 6 double-strand breaks per lethal hit induced by gamma rays. These results are consistent with the hypothesis that an unrepaired DNA double-strand break is a major lethal event in both cases. The increase in sensitivity to ultraviolet (measured by colony-forming ability) seems linear in the number of bromouracils incorporated (0--20% of the thymines), and the linear relationship is much the same for incorporation in one or in both strands of the DNA double helix.


Aortic collagen biosynthesis during renal hypertension, pregnancy and hypertension during pregnancy in the rat.

The synthesis and deposition of collagen and other proteins has been measured in the aorta of the rat by radiolabeling in short term organ culture. Under these conditions, the synthesis of collagen and other proteins is linear for at least five hours. Autoradiography demonstrates a labeling in all cell layers and protein deposition in the extracellular matrix. Hypertension was induced by renal ischemia using Goldblatt's technique. The synthesis and deposition of collagen was stimulated in aorta from the first week of hypertension and in pregnancy, and was even more increased in hypertensive pregnant animals. Reserpine suppresses the rise in blood pressure in operated animals and prevents these modifications.


Strand breaks and alkali-labile bonds induced by ultraviolet light in DNA with 5-bromouracil in vivo.

Supercircular gamma phage DNA with 10 bromouracils/100 thymine bases, irradiated with 313 nm light in Tris buffer and sedimented on alkaline and neutral gradients, showed 4.6 alkali-labile bonds per true single-strand break, in agreement with Hewitt and Marburger (1975 Photochem. Photobiol. 21:413). The same DNA irradiated in Escherichia coli host cells showed about the same number of breaks in alkaline gradients for equal fluence, but only 0.5 alkali-labile bond per true break. Similarly, E. coli DNA with bromouracil irradiated in the cells showed only 10--20% more breaks when denatured with 0.1 M NaOH than under neutral conditions with 9 M sodium perchlorate at 50 degrees C. These results show that true single-strand breaks occur more frequently than alkali-labile bonds after ultraviolet irradiation of DNA containing bromouracil in cells.


Magnetic anisotropy of egg lecithin membranes.

Magnetic realignment and rotational diffusion of cylindrical egg lecithin vesicles were measured under a phase contrast microscope. The anisotropy of magnetic susceptibility times membrane thickness was calculated from the data for several thin-walled vesicles. The resulting values were assigned to discrete numbers of bilayers. The difference between the susceptibilities parallel and perpendicular to the long axes of the lecithin molecules is deduced to be X parallel - X perpendicular = -(0.28 +/- 0.02) . 10(-8) cgs at 23 degrees C, if a bilayer thickness of 60 A is assumed.


Prostacyclin (prostaglandin I2, PGI2) inhibits platelet adhesion and thrombus formation on subendothelium.

Prostaglandin I2 (prostacyclin, PGI2), a substance synthesized in the wall of blood vessels, has been previously shown to inhibit the aggregation of platelets in stirred platelet-rich plasma. We used a method in which segments of deendothelialized rabbit aorta are perfused at arterial shear rates with human blood and found that both platelet adhesion and thrombus formation on subendothelium was inhibited in blood containing 10 nM PGI2. PGI2 appears to reduce adhesion by inhibiting platelet spreading. These findings suggest that PGI2 could regulate the deposition of platelets on vascular surfaces.


Human IgA1 half-molecules: clinical and immunologic features in a patient with multiple myeloma.

Abnormal IgA1 half-molecules consisting of one heavy and one light chain were found in a patient (N.N.) with typical multiple myeloma. The serum and the urine of this patient contained both 7.0S and 3.9S IgA myeloma proteins. The IgA half-molecules (3.9S) were found to have a molecular weight of 59,000 daltons and were composed of one alpha1 chain of about 40,000 daltons and one light chain of 22,000 daltons. Furthermore, enzymatic degradation suggested that the alpha chain of the N.N. half-molecules had a large deletion in its Fc portion. We suggest that its heavy and light chains were probably bound noncovalently, since the interchains connecting the heavy and light chains of these IgA half-molecules were easily dissociated with 1% SDS and 8 M urea. Cytologic studies identified at least two types of myeloma cells, and it is possible that half-molecule IgA production might result from mutation among the myeloma cells producing whole-molecule IgA.


Hb F production of endogenous colonies of polycythemia vera.

Fetal hemoglobin was studied in endogenous colonies produced in plasma clot and methylcellulose cultures of circulating progenitors from patients with polycythemia vera (PV). Analysis of globin chain synthesis showed that gamma chains constituted from 13% to 42% of the non-alpha chains produced in cultured cells, whereas from 27% to over 50% of the endogenous colonies contained Hb F, as indicated by the fluorescent antibody probe. Since the endogenous colonies in PV cultures originate from the abnormal PV clone, the findings provide direct evidence that a single pluripotent stem cell can have committed progeny that differ in their expressions of the Hb F production program.


The use of lymphocyte phosphoglucomutase as a genetic marker in bone marrow transplant recipients.

Lymphocyte phosphoglucomutase can be used as a genetic marker to document successful engraftment in bone marrow transplant recipients. Two patients who underwent marrow transplantation as a treatment for acute leukemia showed a change into donor-type isozyme pattern.


Biological and clinical relevance of human macrophage migration inhibitory factor (MIF).

The first isolation and characterization of a lymphokine, the human MIF (MSF), and the availability of a specific MIF (MSF)-antibody provide an opportunity to establish a new parameter for detection of cellular immune mechanisms in various clinical situations. The precise estimation of quantitative amounts of MIF by sensitive immunochemical techniques could gain new insights into the pathogenesis of a number of clinical disorders. In addition, the exact detection of quantitative amounts of MIF in body fluids could have prognostic value.


Effects of modulators of arachidonic acid metabolism on the synthesis and release of slow-reacting substance of anaphylaxis.

1 Slow-reacting substance of anaphylaxis (SRS-A) was released in the peritoneum of passively sensitized rats challenged with ovalbumin and from rat isolated peritoneal cells stimulated with calcium ionophore A23187. 2 Both monocytes (macrophages) and mast cells appear to be involved in the synthesis and release of SRS-A. 3 The immunological release of SRS-A in vivo is enhanced by indomethacin and inhibited by dexamethasone, mepacrine, 1-phenyl-3-pyrazolidone (1-P-3-P), and methylimidazole. 4 SRS-A release induced by A23187 in vitro is inhibited by dexamethasone, indomethacin 1-P-3-P, eicosatetraynoic acid (ETA) and 15-hydroperoxy arachidonic acid. The inhibition induced by dexamethasone, indomethacin and 1-P-3-P is reduced by an increase in the calcium concentration from 1 mM to 5 mM, whereas the inhibition induced by ETA is increased. 5 The results suggest that a lipoxygenase is important in the synthesis and release by SRS-A.


Effects of a combined oestrogen-progestin preparation on gastric acid and pepsin secretion, serum gastrin concentration and biliary secretion of bile acids, phospholipids, and cholesterol in the cat.

1 Daily ethinyloestradiol (50 mug) and norethisterone acetate (1 mg) treatment (Minovlar) was investigated on gastric acid and pepsin secretion, and fasting serum gastrin concentration in six conscious female cats prepared with chronic gastric fistulae. The effect on biliary secretion of bile acids, phospholipids, and cholesterol was investigated in three conscious female cats prepared with chronic gastric and intestinal fistulae, and cholecystectomy.2 Treatment for 49 days did not alter the gastric acid or pepsin response to either intravenous pentagastrin infusions or a food stimulus. The fasting serum gastrin concentration remained unaltered throughout the study.3 Treatment for 18 days did not alter the percentage concentration of cholesterol in the bile, but reduced the percentage concentration of phospholipid. This was mirrored by a rise in the percentage concentration of bile acids in the bile. These trends were quickly reversed on cessation of treatment.4 There was no sign of cholestasis associated with the treatment. Intestinal flow remained constant throughout the study, there was no lithocholic acid or other abnormal bile acids detectable in any samples, and there was no change in serum aspartate aminotransferase concentration.5 The results suggest that in female cats, treatment with a combined oestrogen-progestin preparation does not exert any beneficial effects on the aetiology of peptic ulceration through the reduction of acid or pepsin secretion, or the lowering of serum gastrin concentration. The preparation shows a tendency to produce more lithogenic bile, and this may partly explain the greater incidence of gall stones in women on the contraceptive pill.


The extraneuronal accumulation of isoprenaline in trachea and atria of guinea-pig and cat: a fluorescence histochemical study.

1 The Falck-Hillarp histochemical technique was used to locate extraneuronal sites of accumulation of isoprenaline in trachea and atria from guinea-pig and cat. With a tissue exposure time to formaldehyde gas of 3 h, isoprenaline was located as green fluorescence.2 Quantitative microphotometry was used to measure fluorescence intensity within cells in the trachealis smooth muscle and the atrial myocardium of both species.3 After incubation of tissues in 50 muM isoprenaline, specific fluorescence was seen in trachealis smooth muscle of both species and in the atrial myocardium of cat but not guinea-pig. In both species, fluorescence was also seen in the chondroblasts of the tracheal cartilage and in blood vessels in all tissues.4 In trachealis smooth muscle of both species and in cat atrial myocardium, fluorescence brightness, resulting from incubation of tissues in 50 muM isoprenaline was significantly increased by 200 muM beta-thujaplicin, an inhibitor of catechol-O-methyl transferase (COMT). In the presence of beta-thujaplicin, fluorescence was not visible in guinea-pig atrial myocardium with 50 muM isoprenaline, although fluorescence brightness measured in myocardial cells was now greater than that in corresponding controls.5 The fluorescence intensity seen in cat and guinea-pig trachealis smooth muscle cells and in cat atrial myocardial cells after incubation in 50 muM isoprenaline was decreased significantly in the presence of phenoxybenzamine (100 muM). In guinea-pig atria, phenoxybenzamine had no effect on myocardial fluorescence. Fluorescence intensity was also decreased if the incubation with isoprenaline was carried out at 0 degrees C or if the post-incubation washing temperature was 37 degrees C instead of 0 to 2 degrees C.6 The results demonstrate that the fluorescence histochemical technique can be used to locate isoprenaline in tissues. They also indicate that guinea-pig and cat trachealis smooth muscle cells and cat atrial myocardial cells can accumulate isoprenaline (a) by a mechanism sensitive to phenoxybenzamine and (b) into sites in which COMT plays a functional role in inactivating isoprenaline at the concentration used in these histochemical experiments (50 muM). In contrast, the guinea-pig atrial myocardial cells may have a minimal capacity to accumulate isoprenaline by a phenoxybenzamine-sensitive uptake mechanism.


A controlled comparison of simulated and real ECT.

Two groups of 16 patients with depressive psychosis took part in a controlled evaluation of electro-convulsive therapy (ECT). One group received six brief pulse unilateral shocks under conventional anaesthesia and muscle relaxation; the second group underwent the same procedure without receiving shocks. Outcome was assessed by a separate investigator using the Hamilton Rating Scale for Depression under double-blind conditions. The results showed that this form of ECT was only superior to the control treatment for one item in the scale, a finding which could have occurred by chance. The results suggest that the ECT pre-treatment procedure has an important therapeutic effect. This casts some doubt on current views of the effectiveness of electro-convulsive therapy in general, and of brief pulse unilateral ECT in particular.


Cyproheptadine in anorexia nervosa.

In three hospitals 81 female patients satisfying rigorous diagnostic criteria for anorexia nervosa were randomly allocated to one of four treatment combinations of cyproheptadine and placebo with behaviour therapy and no behaviour therapy. Cyproheptadine was found to be effective in inducing weight gain in a subgroup of anorexia nervosa patients who (a) had a history of birth delivery complications, (b) had lost 41-52 per cent weight from norm and (c) had a history of prior outpatient treatment failure. This subgroup may represent a more severe form of anorexia nervosa.


Limitations of the single sample tracer method for determining glomerular filtration rate.

Apparent volumes of distribution of 51Cr-EDTA and 99Tcm-DTPA were calculated three hours after intravenous injection and correlated with simultaneous estimation of GFR. A reasonable correlation between these values was obtained for those patients with GFR greater than 50 ml/min (standard error of estimate using 51Cr-EDTA 7.6 ml/min and 99Tcm-DTPA 13.7 ml/min). Theoretical reasons are given why a single sample technique will not be accurate over a wide range of renal function.


Possible effect of time on renal allograft rejection.

The change in plasma creatinine concentrations from decreasing values after successful renal transplantation to increasing values after the onset of rejection occurs as a sudden event. Twenty-two such episodes in 16 renal allograft recipients were studied by extrapolating sequential measurements of plasma creatinine concentrations to see when the change occurred. Seventeen of the episodes occurred between 2300 and 1100 and the rest at other times. This difference was significant. The results suggest that rejection is more common at night and apparently has a circadian rhythm, being likely to first influence creatinine clearance at around 0600.


Bacteriological quality control in human milk-banking.

The bacteriological quality of pooled human milk donated to the Oxford milk bank was analysed and the effects on bacteriology of sterilisation of the milk-collecting vessels in the home with hypochlorite solution and of Holder pasteurisation in a purpose-built human-milk pasteuriser were studied. Collecting milk in hypochlorite-sterilised vessels resulted in a significantly lower bacterial count of both pathogens and species of unlikely pathogenicity before pasteurisation and significantly increased the chance of pasteurisation giving a sterile product. Potentially pathogenic organisms grown in untreated milk were Escherichia coli, Staphylococcus aureus, and group B beta-haemolytic streptococci. Seven species of organisms of unlikely pathogenicity were also identified. Pasteurisation eliminated all potential pathogens from milk but did not reliably remove any of the species of unlikely pathogens. Banked human milk may be contaminated with bacteria which are known to be capable of producing lipases, proteases, and decarboxylases. Accurate pasteurisation, together with attention to the sterility of the collecting vessels, results in a bacteriologically safe product that retains many of the protective properties of raw milk.


Value of examining buffy coats for intragranulocytic micro-organisms in patients with fever.

To determine the significance of the presence of intragranulocytic micro-organisms in the blood buffy coat in patients with suspected infection, buffy coat examination and blood cultures were simultaneously performed in 455 consecutive patients with fever. There was no general correlation between the finding of intragranulocytic micro-organisms in the buffy coat and positive blood cultures. Patients with persistent bacteraemia and sterile blood cultures were, however, shown to have persistently positive buffy coat findings on repeated examination. These patients, who had culture-negative endocarditis or catheter-associated infections, had sterile blood cultures because of antibiotic treatment. Repeated positive findings in the buffy coat may therefore be valuable in detecting patients with persistent bacteraemia, but sporadic findings of micro-organisms in the buffy coats of acutely ill patients seem to have little diagnostic value.


Cardiac transplantation in 150 patients at Stanford University.

A total of 150 patients underwent cardiac transplantation at Stanford between January 1968 and August 1978. Sixty-two patients were alive at the end of this period, when the expected one-year survival rate was 70%, with an attrition rate of roughly 5% thereafter. Changes in postoperative managment resulting from continuing research contributed to improved survival rates. The results are equal or superior to those for recipients of renal transplants from unrelated donors. Major centres that have suitably trained staff and appropriate facilities for cardiac surgery should perhaps reconsider cardiac transplantation as an alternative to death in certain patients.


Malaria in Birmingham and a London teaching hospital.

During the past five years the incidence of imported malaria increased among patients seen in East Birmingham Hospital and in St Thomas's Hospital, London. Plasmodium vivax was the predominant species in Birmingham, and was almost always acquired by Asian immigrants visiting the Indian subcontinent. In St Thomas's P falciparum was most commonly imported, usually by African immigrants visiting Nigeria and Ghana. Two patients (one Irish, one Japanese) died of falciparum malaria after visiting tropical Africa. In both hospitals the immigrant patients had seldom taken prophylactic drugs, and the few who had, ceased to do so on arrival in the UK and sometimes before leaving the malarious country. Apparently immigrants who visit their homeland do not consult their general practitioners before travelling, are given inappropriate advice, or do not take appropriate advice when given. Since the incidence of imported falciparum malaria in the UK is rising, the following points should be considered: the infection may be lethal, particularly in patients lacking immunity; it can mimic other diseases, which may lead to delayed diagnosis; severe disease may be associated with few parasites on a blood film, and even if the result is negative further tests should be performed; clinicians and hospital pharmacists should be aware of the need to keep permanent stocks of parenteral chloroquine and quinine preparations.


Randomised controlled trial of vancomycin for pseudomembranous colitis and postoperative diarrhoea.

The efficacy of vancomycin in pseudomembranous colitis was assessed in a prospective randomised controlled trial. Forty-four patients with postoperative diarrhoea were allocated to five days' treatment with either 125 mg vancomycin six-hourly or a placebo. Sixteen patients had high titres of the neutralised faecal toxin characteristic of pseudomembranous colitis; nine received vancomycin and seven placebo. At the end of treatment faecal toxins were present in one patient given vancomycin compared with five of the controls. Vancomycin caused the disappearance of Clostridum difficile from the stool in all except one patient, whereas toxicogenic strains of Cl difficile persisted in all but one of the controls. Histological evidence of psuedomembranous colitis had disappeared by the end of treatment in six out of seven patients given vancomycin compared with only one out of seven patients given vancomycin compared with only one out of five patients given placebo. In patients with faecal toxins bowel habit had returned to normal in seven of the vancomycin group compared with only one of the controls, but there was no significant difference in clinical response among patients without faecaal toxins. The results suggest that vancomycin eliminates toxin-producing Cl difficile from the colon and is associated with rapid clinical and histological improvement in patients with pseudomembranous colitis.


Treatment and survival on advanced breast cancer.

The final results of a clinical trial comparing endocrine with cytotoxic drug treatment for advanced breast cancer were analysed. Although cytotoxic treatment gave a significantly higher response rate with a remission duration comparable to that obtained with endocrine treatment, the sequence in which the two treatments were given did not appear to influence survival--except possibly in women with rapidly progressing disease, when cytotoxic treatment is preferred.


New approach to treatment of recent stroke.

Ninety-one patients with acute stroke participated in a double-blind, placebo-controlled trial of naftidrofuryl. Treatment was allocated at random and given over 12 weeks, neurological and neurophysical scores being obtained before treatment and at weeks 2, 4, 8, and 12. Both treatment groups greatly improved over the 12 weeks, but the naftidrofuryl-treated patients made greater neurological progress. Of the patients eventually discharged, those given naftidrofuryl spent only half as long in hospital as the controls. Deaths attributable to stroke were significantly fewer in the active-treatment group.


Treponema pallidum immune adherence test for serodiagnosis of syphilis. 1: An improved method of the TPIA test.

Clinical investigations using the Treponema pallidum immune adherence (TPIA) test as reported by Nelson (1953) were performed only by Olansky et al in 1954. In this paper an improved method of the TPIA test is described. The test was simple to perform and was highly specific for antitreponemal antibody. Furthermore, as it was possible to use heated T. pallidum as antigen the antigen suspension could be preserved for long periods. The TPIA test for syphilis was evaluated by examining the sera from 40 healthy subjects and from 166 patients with syphilis.


Treponema pallidum immune adherence test for serodiagnosis of syphilis. 2: Comparison with glass plate, TPHA, and FTA-ABS tests.

Using sera from 340 patients with syphilis the Treponema pallidum immune adherence (TPIA) test was compared with the glass plate, T. pallidum haemagglutination (TPHA), and fluorescent treponemal antibody-absorbed (FTA-ABS) tests. The results of the TPIA test agreed with those of the glass plate, TPHA, and FTA-ABS tests in 65%, 82%, and 73% of cases respectively. In the quantitative TPIA test no significant correlation with the other tests was observed, and it is, therefore, concluded that the TPIA test has highly individual characteristics. From gel filtration the particular feature of the test was its high sensitivity to the IgM antibody. The TPIA test thus appears to be suitable for estimating antibody in the early stages of the disease.


Treponema pallidum immune adherence test for serodiagnosis of syphilis. 3: Clinical significance and evaluation of treatment.

Antibody titres were measured in patients with clinical syphilis, and the effect of treatment on the results of the Treponema pallidum immune adherence (TPIA) test is reported. In the Treponema pallidum haemagglutination (TPHA) test little change in antibody titre occurred after treatment while in the fluorescent treponemal antibody-absorbed (FTA-ABS) test only a slight decrease occurred. The decrease in the antibody titre in the TPIA test was similar to that in the glass plate test, but the findings were different. From the results of gel filtration of sera obtained from patients with syphilis at different intervals after treatment it was apparent that the decrease in antibody titre after treatment mainly concerned the IgM antibody; thus, because of the high sensitivity of the TPIA test to IgM antibody the test is useful in evaluating the effect of treatment.


Rapid diagnosis of chlamydial infection of the cervix.

A rapid serodiagnostic test for the presumptive diagnosis of chlamydial infection of the cervix has been developed. The method used in based on the modified micro-immunofluorescence test using pooled chlamydial antigens and the detection of different immunoglobulin classes of chlamydial antibody in sera and cervical secretions. The presence of IgG chlamydial antibody at a level of 1/64, or IgM antibody at a level of 1/8 or greater, or both in sera and IgG or IgA antibody at a level of 1/8 or more or both in cervical secretions was closely associated with the isolation of Chlamydia trachomatis and non-specific genital infection. In general, serodiagnosis was three to nine times more sensitive than cultural methods, and the detection of IgG chlamydial antibody in cervical secretions alone provided the most sensitive of the serological tests. This sensitive, low-cost, rapid, and simple serodiagnostic test for the presumptive diagnosis of chlamydial infection of the cervix, coupled with transportation of specimens by post, offers advantages over conventional isolation techniques for the routine diagnosis and management of chlamydial genital infections.


Diagnostic, treatment, and reporting criteria for non-specific genital infection in sexually transmitted disease clinics in England and Wales. 1: Diagnosis.

The current methods of diagnosis for non-specific genital infection (NSGI) in clinics in England and Wales are described. In most clinics (92%) microscopical findings were used by consultants to establish the diagnosis of non-specific urethritis (NSU) in male patients. However, the microscopical criteria that they used in reaching a diagnosis varied between clinics. The most commonly applied criterion was that of less than five leucocytes per high power field. NSGI in female patients and non-specific proctitis in passive homosexuals were recognised as distincy in 60% of clinics and the latter in 57%. Among those who recognised these conditions the diagnostic criteria varied. The establishment of acceptable and uniform criteria for diagnosis are discussed.


The concept of a sensorimotor cortex: its early history, with especial emphasis on two early experimental contributions by W. Bechterew.

A brief account has been given of the events during the last quarter of the nineteenth century which led to the concept of the sensorimotor cortex. An early review by Frederick Mott has served as a useful guide. Particular attention has been paid to two early startlingly prescient papers by W. Bechterew. Since, however, Bechterew changed his views in subsequent publications, he forfeited a major claim for priority, and his early papers were forgotten.


Immunocytochemical localization of dopamine-beta-hydroxylase in rat locus coeruleus and hypothalamus.

Dopamine-beta-hydroxylase (DBH), the enzyme that converts dopamine to norepinephrine, has been localized in light and electron microscopic preparations of rat brain by an immunocytochemical method using a peroxidase--anti-peroxidase Fab complex. In light microscopic preparations, DBH-specific reaction product was observed in somata and proximal processes of neurons in the locus coeruleus and subcoeruleus as well as within distal axons of the principal adrenergic fiber system. DBH-specific reaction product was also observed within small (1--2 micrometer), punctate structures in the interstitial nucleus of the stria terminalis and the para- and periventricular nuclei of the hypothalamus. Electron microscopic results demonstrated on association of DBH-specific reaction product with the Golgi apparatus of neuronal somata in the locus coeruleus and subcoeruleus. DBH-positive reaction product was also seen in association with small (35-55 nm) agranular synaptic vesicles and large (80--100 nm), probable granular vesicles within axonal varicosities and terminals in the interstitial nucleus of the stria terminalis. Occasionally, DBH-containing axonal varicosities and terminals were observed to form synapse-like junctions with dendritic profiles, but most of the observed DBH-positive axonal structures did not establish identifiable synaptic relationships.


A proposed cellular mechanism for calcium transport in the intestinal epithelial cell.

Intracellular transport of calcium from the apical to the basal-lateral region of the intestinal epithelial cell was investigated in duodenum from normal fed, fasted, and calcium-loaded rats. The process was followed with time using electron microscopy with potassium pyroantimonate to precipitate calcium. The observations made were subjected to morphometric analysis. The specificity of the method was demonstrated in the villus cell by resistance to microincineration and by absence of deposits following exposure to EGTA. Using this method calcium was seen in cells from calcium-fed rats at the microvillus border, in the Golgi zone, and within the internal compartments of the mitochondria. In cells from fasted rats calcium was not seen. Mitochondria were found largely at the apex of the cell and were free of detectable calcium. By 5 min, in the cells of fasted rats given a calcium load, the calcium had reached the Golgi apparatus and the inner mitochondrial compartment. After 15 min mitochondria were heavily loaded with calcium and had moved to the basal region of the cell. These observations suggest that mitochondria play an important role in absorption of calcium and appear to transport this ion from the apex to the basal region of the cell where entry into the capillaries takes place.


[Non-cardiac acute fulminating pulmonary edema occurring at the end of anesthesia].

We have presented a case of acute fulminating pulmonary oedema, not cardiogenic in origin, occurring at the end of anaesthesia. This precise diagnosis was made with the help of measurement of the capillary wedge pressure and cardiac output as well as analysis of the alevolar oedema fluid. The haemodynamic values and the gas exchange, as measured under various ventilatory patterns, confirms the usefulness of continuous positive pressure ventilation (CPPV). In the genesis of acute pulmonary oedema as described, the role of hypoxia and acute obstruction of the upper airways as presented by the patient is discussed in the light of the present proposed theories.


Bis-intercalative binding to DNA of novel bis(10-methyl)acridinium chlorides and its dependence on chain length of linker.

The synthesis of a series of novel bis(10-methyl)acridinium compounds (both unsubstituted and the 6-chloro-2-methoxy substituted) linked by methylene bridges of lengths from (CH2)4 to (CH2)12 and in one case by spermine is described. Their ability to bind to duplex DNA was compared by their relative inhibition of E. coli DNA polymerase catalyzed DNA synthesis. It was determined that they function as DNA template inhibitors and do not affect the DNA polymerase directly. Their ability to function as bis-intercalators was assessed by a novel and convenient topoisomerase fluorescent assay. It was concluded that whereas the (CH2)4-linked compounds act only as monofunctional intercalators because of steric constraints the (CH2)6-, (CH2)8-, and (CH2)10-linked substituted bisacridinium compounds, as well as the (CH2)10- and (CH2)12- unsubstituted analogues, function as bis-intercalators with DNA.


The viscosity and lipid composition of the plasma membrane of multiple drug resistant and sensitive yeast strains.

Four different plasma membrane preparations were isolated from multiple drug resistant and sensitive isolates of two isogenic groups of Saccharomyces cerevisiae strains: zymolyase ghosts, concanavalin A ghosts, pH 4 nonaggregated vesicles, and sucrose-gradient purified vesicles. The viscosities of these preparations were determined by the use of a fluorescence polarization technique with 1,6-diphenyl-1,3,5-hexatriene. The viscosities of all four membrane preparations within an isogenic set were the same for resistant and sensitive strains. A comparison of the viscosity of zymolyase ghost liposomes showed that zymolyase ghost (glyco) proteins of resistant and sensitive strains had the same effect on viscosity. There was no difference between resistant and sensitive isolates in the mole concentration of the following lipid classes extracted from zymolyase ghosts: phospholipid, sterol, sterol ester, triglyceride, diglyceride, and free fatty acid. The fatty acid distribution of esterified and free fatty acids and the distribution of nine phospholipids was the same in zymolyase ghosts from sensitive and resistant strains. It was concluded that multiple drug resistance does not result from an alteration in plasma membrane viscosity or lipid composition.


The high-risk cardiac patient undergoing general surgery.

The challenge of the high-risk cardiac patient undergoing general surgery will be met only when an aggressive approach is taken to avoid an unfavourable balance between the oxygen supply and demand of the myocardium. In the past this challenge has been accepted in the operating room but postoperative care has been less than adequate. The intelligent use of potent, effective pharmacologic agents and intensive monitoring of myocardial performance intra- and postoperatively have greatly reduced morbidity and mortality in patients with ischemic heart disease undergoing aortocoronary bypass procedures; they can achieve similar results in such patients who must undergo general surgery.


Abdominal trauma with special reference to hepatic trauma.

Of 882 patients admitted over a 6-year period to the adult trauma service at the Health Sciences Centre in Winnipeg, 325 (37%) suffered from abdominal trauma; 241 (74%) sustained a blunt injury and 84 (26%) a penetrating wound. Laparotomy is indicated when there is evidence of intra-abdominal hemorrhage, perforation of a viscus or penetration of the peritoneum. Peritoneal lavage is extremely useful in the diagnosis of intra-abdominal bleeding; it was used in 79 cases among which were four false-positive and two false-negative results. Fifty-six cases of hepatic injury were treated; 43 of these were caused by blunt trauma. Temporary packing and drainage sufficed as treatment in many of these patients but the placing of suture-ligatures at exposed bleeding points is often called for. Resection of hepatic tissue was required in 10 cases and ligation of the right hepatic artery in 2. Of the seven deaths in the series only one was attributed to the liver injury itself.


Management of patients with lung contusion.

Pulmonary contusion represents a serious complication of chest trauma. Its management should include adequate oxygen therapy and ventilatory support. Monitoring of the patient's condition should be by repeated measurement of arterial blood gases; use of the Swan-Ganz catheter is seldom necessary. Broad-spectrum antibiotics are generally indicated but cortisone, cardiotonic drugs and diuretics are not usually required.


Gastrointestinal complications of renal transplantation. 1. The upper gastrointestinal tract.

In 95 consecutive cases of cavaderic renal transplantation followed up for 1 to 83 months (mean 23.1 months) 17 complications developed in the upper gastrointestinal tract of 15 patients; these included duodenal ulcer in 12 and gastric ulcer, esophagitis, hemorrhagic gastritis, small-bowel obstruction and small-bowel perforation in 1 each. The occurrence of a complication was not related to the patient's age, sex, blood group or use of cigarettes or alcohol, the duration of hemodialysis before transplantation, the tissue match or the number of infusions of immunosuppressive medication. One patient died of the complication. The peptic ulcers that developed after transplantation were successfully managed conservatively in 69% of cases. Since surgical treatment in patients whose immune response has been suppressed is associated with an increased frequency of complications such as disruption of suture lines, it is preferable to reserve it for those in whom complications develop that are unresponsive to conservative measures.


Gastrointestinal complications of renal transplantation. 2. The colon.

In 95 consecutive cases of cadaveric renal transplantation followed up for 1 to 83 months (mean 23.1 months) seven colonic complications developed in seven patients; these included ischemic colitis in three, colonic perforation in two, fecal impaction in one and appendicitis in one. Except for appendicitis all the complications occurred within 2.5 months of transplantation and were not related to the patient's age, sex, blood group, or use of cigarettes or alcohol, the duration of hemodialysis before transplantation, the tissue match or the number of infusions of immunosuppressive medication. Two patients died, but not of the complication. In the management of free colonic perforation prompt resection or exteriorization, with avoidance of intraperitoneal suture lines, and continuous postoperative peritoneal lavage may be lifesaving. Early surgical intervention and creation of a colostomy in one of the cases of ischemic colitis proved helpful.


Percutaneous needle aspiration biopsy.

This paper reviews the history, technique, applications, advantages, disadvantages and complications of percutaneous needle aspiration biopsy. This technique, particularly when performed with a time needle (21-gauge or less), is a relatively painless, inexpensive and safe method of obtaining a pathologic diagnosis, and it can often be carried out at the bedside or in the outpatient department. It complements other methods and may obviate, but never precludes, subsequent excisional biopsy. Its advantages are insufficiently recognized. Considerable expertise of the cytopathologist and close cooperation with the clinician are necessary for consistent results.


Hypercalcemia and cancer: an update.

Hypercalcemia is a not infrequent complication of cancer that every physician should be aware of. It is a significant factor in the morbidity and mortality of cancer patients. Almost invariably hypercalcemia is associated with accelerated resorption of bone, which is thought to be mediated by humoral or metabolic factors. Three such factors, parathyroid hormone, E2 prostaglandins and osteoclast activating factor, have been strongly implicated in the pathogenesis of hypercalcemia in cancer patients. Other mechanisms for the hypercalcemia may exist. Accurate diagnosis of the disorder is important in therapy, and current research into the various mechanisms for hypercalcemia in cancer patients may well lead to new modes of therapy that are more specific and perhaps less toxic.


Liposarcomas of the head and neck: a review of the literature and addition of four cases.

Liposarcomas of the head and neck are rare. There have been 25 previously reported patients in the literature. Four patients with head and neck liposarcomas, recently treated by the Head and Neck Surgery Service, Walter Reed Army Medical Center, are presented. Less than half of reported patients were noted to be living without evidence of disease. Prognosis generally corresponds to the cell type of the tumor. Intraoral, cheek and orbital tumors seem to have a worse prognosis compared to neck tumors. Wide local excision remains the treatment of choice. Advances in surgical techniques have allowed adequate therapy for most head and neck liposarcomas. Advanced lesions should be managed by conservative surgery and radical radiation therapy.


Postirradiation sarcoma of bone: a perspective.

Ten cases of irradiation induced sarcoma of bone which fulfilled Cahan's criteria were seen in a twenty year period at the Princess Margaret Hospital. The overall incidence of this complication is 0.035% of all irradiated five year survivors. Combining our data with three other large series presented in the literature, a dose complication curve could be deduced. On the basis of the human and animal data reviewed, it was concluded that the risk of radiation induced sarcoma is so low in the dose range of modern radiotherapeutic practise that it does not represent a contraindication to the use of radiation therapy.


Immunoblastic lymphosarcoma: a light, immunofluorescence, and electron microscopic study.

Immunoblastic lymphosarcoma (ILS) is a newly recognized malignant lymphoreticular neoplasm and is included in the recent W.H.O. classification of lymphomas. This report concerns six cases of ILS studied by light, immunofluorescence (IF), and electron microscopy (EM). Four patients were female and all except one were over 50 years of age. Four patients had some immunological abnormality. Light microscopy showed a monomorphic population of immunoblasts with pyroninophilic cytoplasm and variable plasmacytoid differentiation. Intracytoplasmic IgG was demonstrated by IF in four cases, and IgA in one. Large lymphoid cells with varying proportions of polysomes, rough endoplasmic reticulum, and Golgi apparatus were seen by EM in four cases. Mean survival was 4.8 months in five cases; death in four was due to disseminated ILS. We concluded that our cases of ILS are of B cell origin, are often associated with immunological abnormalities, and carry a poor prognosis. Immunofluorescence and EM are helpful in its diagnosis.


Renal metastases from well differentiated follicular thyroid carcinoma: a case report with light and electron microscopic findings.

The second case of metastatic follicular carcinoma of the thyroid presenting during life as bilateral renal masses is described.  Metastases appeared 18 years after resection of the primary and on clinical evaluation were confined to the kidneys. Although most malignant tumors which present clinically in the kidney are primary there, a review of the literature indicates that the possibility of metastatic tumor must be kept in mind, particularly when the renal tumor is bilateral.


Acute ("malignant") myelosclerosis.

This study is based upon an analysis of the hematologic and pathologic material from seven patients with acute myelosclerosis, as well as a review of the literature of 49 cases reported under this designation, or one of its synonyms. Patients with this disease characteristically present with pancytopenia, minimal or absent anisocytosis and poikilocytosis, and a fibrotic bone marrow showing hyperplasia and immaturity of all three cell lines, with particular prominence of megakaryocytes and their precursors. In addition, clinical splenomegaly is almost always absent, and the disease has a rapidly fatal course. We consider only one-fourth of the cases reported in the literature to have the clinical and hematologic features consistent with the diagnosis of acute myelosclerosis; the remainder represent a variety of myeloproliferative disorders, including chronic myelosclerosis with an accelerated terminal phase, acute myeloblastic leukemia with bone marrow fibrosis, myeloproliferative diseases that cannot be subclassified, and cases in which the data are insufficient for analysis. Using strict clinical and hematological criteria, acute myelosclerosis can be separated from other myeloproliferative disorders as a distinct clinicopathologic entity.


A comparative study of a BCNU containing 4-drug program versus MOPP versus 3-drug combinations in advanced Hodgkin's disease: a cooperative study by the Cancer and Leukemia Group B.

A prospective randomized trial by CALGB examined the relative value of four chemotherapy regimens in 537 patients with stage III B and IV Hodgkin's disease. A new combination BOPP, derived by substitution of BCNU for nitrogen mustard in the MOPP regimen, was compared to MOPP and to two 3-drug regimens, derived by removing the procarbazine in BOPP (BOP) or removing the alkylating agent (OPP). The 4-drug programs gave significantly higher frequency of complete remissions (BOPP 67%, MOPP 63%) than the 3-drug regimens (BOP 40%, OPP 42%), and significantly longer duration of remission and survival. BOPP had a therapeutic activity equal to MOPP, and was accompanied by less toxicity. After 6 cycles of induction chemotherapy, responding patients, both CR and PR, were continued on maintenance chemotherapy for 3 years. No significant difference in relapse rate was demonstrated following maintenance treatment with either vinblastine, chlorambucil, or chlorambucil plus monthly vincristine + prednisone doses. Nor could a reinforcement phase late in the maintenance program be shown to influence the relapse rate. The median survival for all patients entered on the 4-drug programs was 5 years, while the median has not yet been reached at 6 years for those patients, who obtained CR.


Malignant lymphoma simulating leukemic reticuloendotheliosis: a clinicopathologic study of ten cases.

We have studied ten patients with a lymphoproliferative disorder characterized by massive splenomegaly; minimal lymphadenopathy; varying degrees of blood cytopenias; circulating atypical lymphoid cells frequently with "hairy" cytoplasm; monoclonal serum paraprotein; and tartrate-resistant acid phosphatase reactivity in the tumor cells of five patients tested. Although the clinical and laboratory features in most cases prompted a clinical diagnosis of "hairy cell leukemia" (HCL), histologic, ultrastructural and immunohistologic studies of multiple organs revealed distinctive features recognizably different from leukemic reticuloendotheliosis (LRE). Because this B lymphocyte proliferation may be mistaken for LRE in cases where careful histologic study is not performed, it may be responsible in part for the conflicting data in attempts to characterize the cell of origin of the latter disease. Clinical and experimental data in HCL must be questioned if they do not include histopathologic confirmation of the diagnosis.


Immunoblastic sarcoma: a clinical description.

We reviewed the clinical records of 33 patients with Immunoblastic Sarcoma in order to further describe this disease clinically. Several common features were found. Thirty percent of the patients had a history of a prior immune disease or lymphoproliferative malignancy. Forty-four percent of the patients tested had a diffuse hypergammaglobulinemia. Lymphopenia (less than 1,000/mm3) was found in 45%, and anemia occurred in 73%. At initial presentation, 30% of the cases were clinically staged as either stage I or II, whereas 70% were found to be stage III or IV. Forty-nine percent of the patients had systemic symptoms at presentation. The median survival was 14 months. Advanced stage of disease, lymphopenia, and presence of systemic symptomatology were associated with significantly decreased survival times (p less than .05). We conclude that IBS is a clinical entity often associated with prior immune disease and/or diffuse hypergammaglobulinemia.


The triad of gastric epithelioid leiomyosarcoma, functioning extra-adrenal paraganglioma, and pulmonary chondroma.

The triad of gastric epithelioid leiomyosarcoma, functioning extra-adrenal paraganglioma, and pulmonary chondroma was found in two young female Mayo Clinic patients, and two of these tumors occurred in another two. Review of the world literature disclosed nine patients who probably had two or more of the three neoplasms. We are also aware of two other patients with two of the three tumors. The association of gastric epitheloid leiomyosarcoma, functioning extra-adrenal paraganglioma, and pulmonary chondroma may well constitute a syndrome because of 1) the improbability of the coincidental occurrence of this triad of tumors, 2) the multicentricity of the tumors in the organs or system affected, and 3) the development of the tumors at a young age. Because two of the components of the triad are potentially lethal, it is important that patients less than 35 years of age who have any one of the three tumors be examined periodically to search for the others.


Low dose chemotherapy of metastatic breast cancer with cyclophosphamide, adriamycin, methotrexate, 5-fluorouracil (CAMF) versus sequential cyclophosphamide, methotrexate, 5-fluorouracil (CMF) and adriamycin.

Seventy-eight advanced breast cancer patients with hormone-resistant disease or visceral metastases were randomized to receive either of two low dose regimens consisting of cyclophosphamide (C), methotrexate (M), 5-fluorouracil (F), and Adriamycin (A) as their initial chemotherapy. One group was treated with CAMF, and the other with CMF until progression, followed by A (CMF leads to A). C was given at 50 mg/m2, po, days 1-14; M at 20 mg/m2, F at 300 mg/m2, and A at 20 mg/m2, iv, days 1 and 8 of each 28-day cycle. The response rates for CAMF vs. CMF did not differ significantly (complete and partial responses-62% vs. 49%; stabilizations-23% vs. 31%). Responses by site of metasis, median times to progression and median survivals were similar for both groups. Poor and good risk partial responders had similar survivals. Twelve percent of CMF patients treated with Adriamycin at the time of progression had partial responses with an associated improved survival. Since CMF is as effective as CAMF, but has less toxicity, low dose therapy with CMF is more acceptable than CAMF as an initial chemotherapy regimen for metastatic breast cancer. Adriamycin may be reserved for subsequent regression induction.


A randomized study of two different schedules of methyl CCNU, 5-FU and vincristine for metastatic colorectal carcinoma.

Sixty-seven patients with measurable colorectal carcinoma were randomized to receive two different schedules of Methyl CCNU, 5-FU and Vincristine (MOF). The treatment schedule consisted of Methyl CCNU 150 mg/m2 po q 70 days, 5-FU 300 mg/m2 iv for 5 consecutive days q 35 days, and Vincristine 1 mg/m2 IV q 35 days. The same total dose was used in each arm; in MOF A, the Methyl CCNU was given on day 1, while in MOF B, the Methyl CCNU was divided and given over 5 consecutive days. In MOF A, there was a 10% partial response rate and a 10% minor response rate. In MOF B, the partial response rate was 12% and the minor response rate was 21%. This difference is not statistically significant, but the patients in MOF B experienced less gastrointestinal toxicity (p less than .001).


Chemotherapy versus chemoimmunotherapy in advanced adenocarcinoma of the colon and rectum: a prospective randomized study.

The combination of vincristine, methyl-CCNU, and methotrexate with or without MER-BCG achieved a 2% complete response (CR) and a 11% partial response (PR) with a median duration of 25-29 weeks in 124 evaluable patients with advanced adenocarcinoma of the colon and rectum. Responses were seen in previously untreated patients and in patients refractory to 5-fluorouracil. The median survival of these objective responders (CR + PR) was 57 weeks. The addition of MER-BCG did not appear to influence response rate or duration of survival and was accompanied by significant toxicity. Response was significantly correlated with performance status, sex, and disease free interval and survival with alkaline phosphatase and performance status. Patients with advanced colorectal carcinoma should be stratified according to these variables.


Committee on Growth, 1945-1956: another noble experiment.

The events of the creation, activities, and termination of the Committee on Growth of the National Research Council as the advisory body on research to the American Cancer Society, 1945 to 1956, are historical and instructive. It remains to be demonstrated that programmed research support is more effective than the less structured approaches represented by the Committee on Growth.


Banding studies of chromosomal abnormalities in patients with acute lymphocytic leukemia.

Karyotypes were analyzed by routine Giemsa and quinacrine fluorescence for 16 patients with acute lymphocytic leukemia [ten adults (18 to 51 years) and six children (3 to 15 years)]. Four patients had received previous therapy, but all 16 had active disease when they were first studied. Eight patients (five untreated) had a normal karyotype initially; however, three of these developed a chromosomal abnormality during relapse. Eight patients had a chromosomal abnormality in their initial samples. Each of the 11 patients had different abnormalities. All chromosomes except Nos. 3, 5, 15, 16, and Y were involved in the various aneuploidies. One patient had a Ph1 chromosome due to a translocation with No. 21: t(21;22)(q22;q11). A patient with B-cell acute lymphocytic leukemia had a 14q+ marker in addition to other abnormalities. The median survival of patients with initially normal karyotypes may be longer than that of patients whose karyotypes are abnormal initially.


Primary neoplastic transformation in vivo of xenogeneic skin grafts on nude mice.

Rabbit skin was infected with Shope papilloma virus and grafted orthotopically to nude mice. Typical Shope rabbit papillomas developed in most of the grafts, but similarly treated nude mouse skin grafts were not altered. These results demonstrated that xenogeneic tissues retained susceptibility to oncogenic agents after transplantation to nude mice. The results also implied that it should be feasible to study carcinogenesis of human tissues in this system.


Historical background and aspects of the mechanism of leukocyte adherence inhibition.

The development of leukocyte adherence inhibition is traced from early experiments with murine tumors to recent applications in human cancer and in cell-mediated immunity in general. Arbitrary experimental conditions (serum in culture medium, preincubation, and tumor extracts at a single concentration) were initially set up and permitted detection of specific inhibition of adherence of sensitized leukocytes reacting with antigen of the tumor extracts. Blocking serum completely or partially restored adherence. It was readily demonstrated that leukocyte adherence inhibition was mediated by a soluble lymphokine-like factor [leukocyte adherence inhibition factor (LAIF)]. Recent studies suggest that serum is important in enabling detection of LAIF;  this may explain the inability of some other workers to confirm our findings. Serum appears to protect LAIF from enzymatic destruction in mixtures containing tumor extracts. Complex cell interactions are involved in LAIF production. With mouse cells in vitro, macrophages are required for production by both T- and B-lymphocytes, and there is evidence for suppressor cells also.


Evaluation of the microplate leukocyte adherence inhibition test and its reproducibility, sensitivity, and relationship to other tests of cellular immunity.

The microplate version of the leukocyte adherence inhibition (LAI) test was evaluated in murine and human studies. It was used in parallel with the microcytotoxicity assay, lymphotoxin assay, leukocyte migration inhibition, and lymphocyte stimulation tests in a transplantable murine tumor model, and it compared favorably with these established techniques for the detection of cellular immunity. The LAI test detected both primary and secondary anamnestic responses in Bacillus Calmette-Guérin-primed mice, and it displayed sensitivity to host humoral factors comparable to that seen with other tests. The LAI phenomenon was shown to be mediated by a soluble supernatant factor liberated by antigen-exposed immune leukocytes in the mouse and by concanavalin A-stimulated human leukocytes. In the mouse, deliberate depletion of T-cells ablates LAI reactivity in cells taken at the peak of a primary response; however, immune serum "arms" non-thymus-dependent cells taken from unimmunized hosts. In the mouse, the results of LAI tests correlate with other established techniques, when purified protein derivative reactivity was assessed in spleen cells from Bacillus Calmette-Guérin-immunized mice. Comparable correlation is not found when reactivity to this antigen is assessed in human peripheral blood samples from unimmunized donors.


Correlations between the leukocyte adherence inhibition microassay and in vivo tests of transplantation resistance.

The leukocyte adherence inhibition (LAI) microassay detects tumor-associated antigen(s). Extracts of colon carcinoma (MCA-38 and B16 melanoma tumors, both syngeneic to the C57BL/6J mice) are recognized only by peritoneal cells from mice bearing the corresponding tumor. To ascertain whether this in vitro antigenic recognition correlates with the ability of the host to recognize and reject a tumor in vivo, serial LAI microassays were performed synchronously with experiments designed to test the ability of mice bearing tumors to reject live secondary tumor challenges. Concomitant tumor immunity was present in the MCA-38 tumor-bearing mice on 3 occasions from 5 to 15 days from primary inoculation. In the B16 system, concomitant immunity was present on one occasion 10 days after primary inoculation. These results in turn were paralleled with the specific in vitro recognition of tumor antigens as detected by the LAI microassays. Loss of immunity in the "eclipse" phase of tumor development, as detected by concomitant tumor immunity, was paralleled by nonreactivity of the indicator cells in the LAI microassay.


Cellular and humoral factors involved in the mechanism of the micro-leukocyte adherence inhibition reaction.

To study the cellular basis for specific antigen-induced leukocyte adherence inhibition, enriched populations of B-cells, T-cells, and monocytes were prepared by a two-stage adherence separation procedure from spleen cells of normal C57BL/6J mice and mice bearing progressively growing MCA-38 tumors. The reactor cell undergoing specific antigen-induced adherence inhibition was identified as a monocyte (esterase positive, did not respond to mitogens, and did not bear Thy 1.2 antigen or surface immunoglobulin). Furthermore, an enriched population of MCA-38-sensitized B-cells could program normal monocytes to undergo specific antigen-induced adherence inhibition. In contrast, enriched populations of MCA-38-sensitized T-cells could not program normal nylon wool-adherent cells to undergo antigen-specific adherence inhibition. Programming of normal monocytes by MCA-38-sensitized B-cells occurs through a soluble mediator and not by direct cell contact. The soluble mediator appears to be immunoglobulin in nature and induced both adherence inhibition and the inhibition of adherence. Thus, in this murine tumor model, leukocyte adherence inhibition appears to be due to programming of monocytes by a secretory product of specifically sensitized B-cells.


Assessment of the mechanism of the leukocyte adherence inhibition test.

This study was designed to elucidate the mechanism of the leukocyte adherence inhibition (LAI) test in man. To identify the reactive cell types, enriched leukocyte populations (dextran-separated leukocytes and Hypaque-Ficoll-isolated mononuclear cells and neutrophils, as well as rosette-isolated B- and T-lymphocytes) were tested for leukocyte adherence in the absence of serum to tumor-specific antigens. LAI reactivity was not restricted to any of the enriched populations, suggesting the involvement of multiple cell types. Attempts to demonstrate soluble lymphocyte factors in the LAI mechanism have been uniformly negative. In contrast, factors in serum of immune donors were able to arm naive cells to be specifically responsive. This suggests a role for serum factors in the mechanism of LAI reactivity and partially explains the participation of multiple cell types in the responses observed. In additional studies, we could not document a correlation between the magnitude of the dermal test (delayed cutaneous hypersensitivity) and the magnitude of the LAI response in patients with squamous cell carcinoma of the head and neck. In 34 of 54 of these patients, there was agreement between the two tests (both positive, 27 of 54; both negative, 7 of 54). In the remaining 20 patients, the dermal test was greater than 5 mm while the LAI test was negative (less than 30% inhibition).


Demonstration of the microtest version of the leukocyte adherence inhibition assay.

This paper gives a detailed description of the microtest leukocyte adherence inhibition technique, as well as the results obtained with blood specimens coded by impartial observers. Three coded blood specimens from patients with colon cancer, pancreatic cancer, and melanoma were tested against crude membrane preparations of pancreatic and colon adenocarcinoma tumors. No tissue type-specific reactivity was observed. The inability to demonstrate specific reactivity was due to extensive variability observed within each test. The extensive variability resulted from time constraints of the workshop that necessitated deviations from the normal procedure.


Malignant pleural effusions.

Recurrent malignant pleural effusion is a relatively common problem that often occurs many months before the terminal stages of a patient's malignant disease. Despite careful evaluation, it is often impossible to identify the exact physiologic cause of an effusion and difficult to identify a pragmatic course of therapy. Aggressive management including chest tube drainage, with or without instillation of inflammation-inducing drugs, and radiotherapy or surgery in selected cases is indicated for palliative treatment and probably for increased survival. In formulating a treatment plan, attention should be given to a variety of factors including pathogenic forces in the formation of malignant pleural effusion, tissue type of the metastatic malignancy, general clinical status of the patient, the presence of pleural loculations and adhesions, and the exclusion of other non-malignant causes of pleural effusion.