[Pluri-dimensional study of psychological factors in coronary disease]. To palliate certain criticisms levelled at workers studying psychological factors in coronary artery disease, the authors have applied a pluri-dimensional approach associating a semi-direct psychological interview, a self-evaluation test (Bortner scale), an Eysenck personality test and Sandler and Hazari's test of obsessional behaviour. This protocol was applied to 222 patients hospitalised for coronary artery disease and 522 random controls. The psychological interview and Bortner test showed a significantly higher proportion of A pattern and especially extreme A pattern behaviour in patients with coronary artery disease (32.6 % in coronary patients 9.7 % in controls). The personality questionnaire showed a greater tendency to neurotic behaviour in the coronary patients. These results were independant of age and sex. In the present study, psychological factors are given equal importance to other major risk factors (tobacco, hypertension, hypercholesterolaemia). They are independant of these other factors. A prospective study using the same protocol is being prepared.


[Isoproterenol test in coronary failure. Comparisons with exercise stress and coronary arteriography]. The results of Isoprenaline stress tests, exercise electrocardiography and coronary arteriography were compared in 90 patients suspected of having coronary insufficiency. The technique used was a continuous intravenous infusion starting with an initial dosage of 3.8 gamma/minute and increasing progressively until the heart rate was equal to or greater than 130/minute, and sustained for three minutes. The same criteria of "positivity" were used as in the exercise test but only electrical charges which persisted or appeared 3 minutes after stopping the infusion were taken into consideration. In this series the sensitivity and specifity of the Isoprenaline test was greater than the exercise test if coronary arterial narrowing equal to or greater than 50% is considered significant. The prognostic values of positive tests which are classified in three groups according to the severity of the coronary artery disease, and the value of the isoprenaline test in the diagnosis of moderate single vessel disease are emphasised.


[Treatment of early post-infarction ventricular aneurysms by assisted circulation and surgery]. Ventricular aneurysm formation in the 3 months following transmural myocardial infarction is rare but may cause serious complications. Cardiac failure and/or ventricular arrhythmias resistant to medical treatment are indications for ventricular resection. The operative mortality is high in this group of patients. 8 patients with ventricular aneurysms of average volume (124 +/- 117 ml/m2) and very impaired left ventricular function (EF : 21 +/- 10%, akinesia : 53 +/- 10%) were operated on with two early deaths and one death in the 7th post operative month. The long term clinical result was satisfactory in the surviving patients, and confirmed by haemodynamic investigation in two of them. The benefical effects of intra-aortic balloon pumping, used preoperatively in all patients, and associated myocardial revascularisation procedures performed in some of them are discussed.


[Change in heart function in Duchênne de Boulogne myopathies at the severe progressive stage]. Left ventricular function was studied in 15 patients with rapidly progressive Duchenne de Boulogne muscular dystrophy. Only one patient had a previous history of cardiac failure. All patients had typical electrocardiographical changes. Haemodynamic and angiographic investigation was performed in all patients. Three groups of patients were distinguished according to angiographic criteria : group I comprising 7 normal patients. group II comprising 4 "intermediary" patients, and group III of 4 patients with hypokinetic cardiomyopathy. Segmental ventricular dyskinesis was observed in 6 patients. Mitral incompetence was present in 2 cases and mitral valve prolpse in 1 case. All patients had a normal resting cardiac index. Patients in group II and III had end diastolic left ventricular pressures greater than 15 mmHg. An intravenous angiotensin test was performed in 4 patients from groups I and II. Impaired left ventricular function was demonstrated in patients thought to be normal in the basal state. Finally, myocardial involvement in this group of patients runs a parallel course to the duration and severity of the peripheral muscular disease.


[Study of hemodynamic effects of the methoxamine in 8 cases of obstructive cardiomyopathy with mitral valve insufficiency]. The haemodynamic effects of intravenous Methoxamine were studied in 8 patients with hypertrophic obstructive cardiomyopathy (HOCM) with mitral incompetence (group I) and 8 patients with mitral incompetence due to valvular, subvalvular or other causes (group II). In both groups the left ventricular end-diastolic pressure rose significantly without any great change in the end diastolic surface area: the ventricular distensibility is reduced in both cases. The mean capillary pressure, the V wave/mean capillary pressure ratio, and the degree of regurgitation of the two groups changed in opposite ways : in mitral incompetence due to varying causes (group II) the regurgitation increased, but in mitral incompetence due to HOCM (group I), it decreased. The left ventricular intracavitary gradient in HOCM tended to disappear under the effect of Methoxamine as the systolic aortic pressure approached the basal values recorded in the apical area.


[Electrophysiological study on mexiletine in man with reference to dose-response relation]. The electrophysiological properties of Mexiletine were investigated by endocavitary His Bundle recording and programmed electrical stimulation of the heart in 30 patients. Differing dosage (2, 2.7 and 3.4 mg/kg) were given intravenously in 3 groups of 10 patients. The effects on the length of the sinus cycle, conduction intervals and cardiac refractory periods were observed and the following results obtained : 1. Sinus rhythm increased at all dosages but this effect was much more pronounced with 3.4 mg/kg dosage (--12.8% +/- 2.81% : : % shortening of sinus cycle with respect to the basal cycle +/- SD, p less than 0.005); 2. Atrioventricular nodal conduction time (A-H interval) decreased, the effect being more marked with the larger dose regimes; 3.His-Pirkinje conduction time (H-V interval) unaltered except in 3 cases where it increased by 5 ms after injection of 2.7 mg/kg; 4. Relative refractory period of His-Parkinje system shortened, this effect also being more pronounced with the larger doses ( --3.75 +/- 0.25% :2.7 mg/kg, p less than 0,001; -- 7 +/- 1.46% 3.4 mg/kg, p less than 0.005). In conclusion, the changes observed in the His-Purkinje system after mexiletine were similar to those of Lignocaine and Diphenylhydantoin. The drug also appears to have a marked vagal inhibitory effect as shown by the acceleration of the sinus rhythm and reduced atrioventricular conduction times.


[Duodenal bacterial flora in the child during post-surgical stasis]. The bacterial flora of the small bowel was studied in infants with stasis after surgery. Samples were obtained, with radiological assistance, by duodenal aspiration and then immediately analysed by the laboratory in blind tests, both qualitatively and quantitatively. There were two groups of patients: one receiving parenteral nutrition and a second on enteral elementary diet; patients without any gastro-intestinal tract disease formed the control group. Pseudomonas aeruginosa and Enterobacteria were isolated significantly more often in both groups of patients compared to the control group. Quantitatively, the predominant flora -- and not total flora -- was different in the three groups. When considering bacteriological species, only P. aeruginosa (greater than 10(6)/ml) differed significantly in numbers in both groups compared to the control group. With these bacteriological investigations, the "contaminated proximal small bowel syndrome" is better defined and a group of high-risk patients may be identified.


[Infantile form of Gaucher's disease. Clinical and biological studies in 1 case. Prenatal diagnosis in 2 further normal pregnancies]. A clinical report of a typical case of infantile Gaucher's disease is given. Study of the beta-glucosidase activity in leukocytes and skin fibroblasts confirmed the diagnosis in the patient and established the heterozygote condition of the parents. Two later pregnancies have been monitored by enzymatic study of cultured amniotic fluid cells. Both foetuses were predicted to be healthy, which was confirmed after birth. The report includes a histopathological ultrastructural, chemical and enzymatic study of liver, spleen and brain obtained at autopsy. Possible clues to the physiopathological mechanism of the cerebral lesion are discussed.


Somatic discomforts among depressed women. An inventory of 69 somatic discomforts was used to identify those discomforts most likely to be concurrent with a clinically severe depression in a sample of 223 recently hospitalized women. The inventory provided scores for each of 15 classes of discomfort. The classes of discomfort with the highest average score for the depressed sample at admission also yielded significantly lower scores for a nonpatient control sample (P less than .05). The four classes of discomfort most pertinent to depression were designated autonomic, wakefulness, dry mouth, and fatigue. The items of discomfort contributing to these classes showed a statistically significant diminution in severity during treatment (P less than .05).


Implications of malnutrition in the surgical patient. The substantial prevalence of malnutrition in the hospitalized patient population has only been recently recognized. Preoperative nutritional and immunological assessment was performed prospectively on admission in 64 consecutive surgical patients. Factors measured included weight loss, triceps skinfold, midarm muscle circumference, creatinine-height index, serum albumin level, serum transferrin level, total lymphocyte count, serum complement level, serum immunoelectrophoresis, lymphocyte T rosettes formation, neutrophil migration, and delayed hypersensitivity. Using these criteria for malnutrition, 97% of the patients had at least one abnormal measurement and 35% had at least three abnormal measurements. Patients were monitored for complications during their hospital course. Serum albumin level, serum transferrin level, and delayed hypersensitivity reactions were the only accurate prognostic indicators of postoperative morbidity and mortality. Substantial unrecognized malnutrition exists in the surgical patient population. An isolated indicator of malnutrition should be interpreted with caution. The visceral protein compartment (serum albumin and serum transferrin levels and delayed hypersensitivity) is the most accurate prognostic indicator of postoperative morbidity and mortality. Perioperative nutritional support may reduce operative morbidity and mortality in the malnourished operative candidate.


[Various immunomorphological tests of cellular immunity in lymphogranulomatosis]. Diffusion chambers were used to study the migration activity and blast transformation of lymphocytes in fragments of lymph nodes from untreated patients with lymphogranulomatosis of various histological variants and stages of the disease. Early in the disease, and particularly at its peak, a progressive decrease in the activity of T-lymphocytes was observed indicating the inhibition of cellular immunity in the patients.


Platelet aggregation in experimental spinal cord injury. Ultrastructural observations. Endothelial alterations occur as early as 1 1/2 minutes following impact injury to the primate spinal cord. Separation of the endothelial junctions and exposure of microvascular basal lamina result in platelet adhesion and aggregations that cover defects in the vessel wall and may progress to complete vascular occlusion. This occurs during the first six hours following injury. Platelets also adhere to the surface of damaged endothelium. Hemostasis resulting from platelet thrombus formation is responsible in part for decreased blood flow in the central gray matter following spinal cord trauma.


The ocular manifestations in Fabry's disease. We present the ocular manifestations in a series of 37 hemizygous male and 25 heterozygous female patients with Fabry's disease. The ocular findings typically do not impair vision, but are unique and diagnostic. Whorl-like corneal deposits were seen in almost all patients and were more severe in the heterozygotes. The lens showed cream-colored anterior capsular deposits, sometimes in striking "propeller" distribution, in one third of the hemizygotes, and in none of the heterozygotes. A faint but unique posterior capsular opacity with a branching radial pattern was seen in 37% of the hemizygotes and 14% of the heterozygotes. Conjunctival vessel aneurysmal dilations and retinal vessel tortuosity were both more frequent and severe in the hemizygotes. Severe visual loss occurred in two hemizygotes as a result of unilateral total central artery occlusions.


Radioactive and bioassay of intraocular antibiotics: double-assay technique to compare penicillin G, cefamandole, and gentamicin in ocular tissues in vivo. We examined the correlation between radioactive assay and trephine-discbioassay of penicillin G sodium, cefamandole nafate, and gentamicin sulfate in ocular tissues of pigmented rabbits after subconjuctival administration of antibiotic. We devised a technique whereby a single sample of tissue could be assayed by both methods. This was achieved by performing the bioassay first, then measuring the resudual radioactivity in the agar and specimens. The results of both methods were generally within 13%. An exception was gentamicin in iris and choroidretina, for which the bioassay result was strikingly less than the radioassay value. No such discrepancy was evident when similar studies were carried out with gentamicin in albino rabbits. This suggests that the phenomenon is due to tight binding of gentamicin by melanin-containing tissues. The trephine-disc bioassay provides an accurate measure of diffusible bioactive antibiotic in ocular tissues.


Local antivirals in a herpes simplex stromal keratitis model. A herpes simplex stromal keratitis rabbit model, which was produced by intrastromal injection of live virus, was used to evaluate the effects of local antivirals on the natural course of the disease. Topical trifluridine (trifluorothymidine) and vidarabine monophosphate (adenine arabinoside monophosphate), when given early and frequently, suppressed the disease, indicating that viral replication was important in initiating the disease. However, seven days after the stromal disease had begun to develop, neither drug had an appreciable effect. Since the early drug effect had suggested adequate drug penetration, the abscence of drug effect later in the disease indicates that viral multiplication may not be important in maintaining the disease. Immunologic reactions may control the disease once the cornea is antigenically altered by the initial infection. Subconjunctivally injected trifluridine was not effective.


Vestibular effects of electrical stimulation of the cochlea. Monitored in the awake primate. The effect of electrical stimulation of the cochlea on vestibular response was monitored in six rhesus monkeys. Eye movements and single-unit activity from the vestibular portion of the eighth nerve, vestibular nuclei, reticular formation, and abducens nucleus were observed while electrical stimulation was delivered through an implanted cochlear prosthesis. In one animal of this series, neural activity from the inferior colliculus and cochlear nuclear complex was also recorded. Electrical stimulation elicited eye-movement responses in only one animal. In the animals from which single-unit activity was recorded, no positive vestibular effects were noted. In one animal of this study, responses were elicited from auditory structures by relatively low intensities of electrical stimulation.


Skin flap esophagostomy. A new procedure. Stoma formation in cervical esophagostomy can be accomplished without tension with the use of cervical skin rotation flaps to form a skin-lined tube. This procedure was performed on six patients; salivary leakage was less, and tube insertion was easier than in patients who had standard esophagostomy procedures. We discuss the indications and contraindications for esophagostomy.


The support of the long term neurological cripple. The term 'long term neurological cripple' is an unattractive and yet an all embracing one, covering a wide spectrum of disorders from spina bifida or cerebral palsy with or without associated epilepsy and behavioural and learning problems, through muscular dystrophy, multiple sclerosis, and motor neurone disease, to the effects of head injury, cerebrovascular lesions and the degenerative disorders of later life such as Parkinson's disease and the senile and presenile dementias. Whilst many of the problems are common to several of these entities, each has its own particular aspects.


Epilepsy.

Epilepsy. Epilepsy is a common disorder affecting approximately one in every two hundred people, from all walks of life, and presenting in addition to the seizure disorder itself, varying social, psychological and economic problems. A simple classification of epilepsy is described, and the concept of 'seizure threshold' introduced. Having diagnosed epilepsy, the need for investigation to separate the symptomatic epilepsies from idiopathic epilepsy is stressed. Accurate diagnosis of seizure type ensures that optimal therapy can be offered. The various drugs used to control epilepsy are discussed, and comments on the general management of the patient are offered.


Inhibition of bacterial transport by uncouplers of oxidative phosphorylation. Effects of pentachlorophenol and analogues in Bacillus subtilis. Analogues of the potent uncoupler of oxidative phosphorylation pentachlorophenol were tested as inhibitors of proline and glycine transport by Bacillus subtilis. These analogues included less highly substituted chlorophenols and pentachlorothiophenol. Like pentachlorophenol, they are non-competitive inhibitors of proline transport and uncompetitive inhibitors of glycine transport. However, the less highly substituted chlorophenols are weaker acids than pentachlorophenol and also weaker inhibitors. Analysis indicated that the anionic form of the uncouplers is the inhibiting species. Pentachlorothiophenol, a water-insoluble anion, is also a potent inhibitor. These results support previous studies that concluded that uncouplers of oxidative phosphorylation inhibit amino acid transport by binding at specific sites on proteins, the free energy of interaction stabilizing 'unproductive' conformations. Such specific interactions of uncoupler with protein are probably commonplace.


Turnover of liver glycogen in the rat foetus. Incorporation and release of the radioactivity in the liver glycogen of 18.5- and 19.5-day-old rat foetuses were studied after intravenous injection of E11-14C]glycerol. Incorporation occurred during 1 h after injection of the radioactive tracer to the foetus; then, the incorporated radioactivity decreased. Glycogen content in the liver, and glycogen phosphorylase and glycogen synthase were not modified during the experiment. It is therefore postulated that a physiological turnover of glycogen exists in the liver of the rat foetus.


Measurement in vivo of hydrogenase-catalysed hydrogen evolution in the presence of nitrogenase enzyme in cyanobacteria. A method was devised that allows measurement in vivo of hydrogenase-catalysed H2 evolution from the cyanobacterium Anabaena cylindrica, independent of nitrogenase activity, which is also present. Addition of low concentrations of reduced Methyl Viologen (1-10mM) to intact heterocystous filaments of the organism resulted in H2 evolution, but produced conditions giving total inhibition of nitrogenase (acetylene-reducing and H2-evolving) activity. That the H2 formed under these conditions was not contributed to by nitrogenase was also supported by the observation that its rate of formation was similar in the dark or with Ar replaced by N2 in the gas phase, and also in view of the pattern of H2 evolution at very low Methyl Viologen concentrations. Conclusive evidence that the H2 formed in the presence of Methyl Viologen was solely hydrogenase-mediated was its evolution even from nitrogenase-free (non-heterocystous) cultures; by contrast 'uptake' hydrogenase activity in such cultures was greatly decreased. The hydrogenase activity was inhibited by CO and little affected by acetylene. Finally the hydrogenase activity was shown to be relatively constant at different stages during the batch growth of the organism, as opposed to nitrogenase activity, which varied.


Interaction of the lacZ beta-galactosidase of Escherichia coli with some beta-D-galactopyranoside competitive inhibitors. 1. The location of the bivalent metal cation with respect to bound competitive inhibitors in Escherichia coli (lacZ) beta-galactosidase was investigated by proton magnetic resonance. 2. Replacement of Mg(2+) by Mn(2+) enhances both longitudinal and transverse relaxation of the methyl groups of the beta-d-galactopyranosyltrimethylammonium ion, and of methyl 1-thio-beta-d-galactopyranoside; linewidths are narrowed by increasing temperature. 3. The Mn(2+) ion is located 8-9A (0.8-0.9nm) from the centroid of the trimethylammonium group and 9A (0.9nm) from the average position of the methylthio protons. 4. The effective charge at the active site was probed by measurement of competitive inhibition constants (K(i) (o) and K(i) (+) respectively) for the isosteric ligands, beta-d-galactopyranosylbenzene and the beta-d-galactopyranosylpyridinium ion. 5. The ratio of inhibition constants (Q=K(i) (+)/K(i) (o)) obtained with 2-(beta-d-galactopyranosyl)-naphthalene and the beta-d-galactopyranosylisoquinolinium ion at pH7 with Mg(2+)-enzyme was identical, within experimental error, with that obtained with the monocyclic compounds. 6. The variation of Q for Mg(2+)-enzyme can be described by Q=0.1(1+[H(+)]/4.17x10(-10))/1+[H(+)]/10(-8)). 7. This, in the theoretical form for a single ionizable group, is ascribed to the ionization of the phenolic hydroxy group of tyrosine-501. 8. The variation of Q for Mg(2+)-free enzyme is complex, probably because of deprotonation of the groups normally attached to Mg(2+) as well as tyrosine-501.


Characterization of a high-molecular-weight glycoprotein isolated from the pulmonary secretions of patients with alveolar proteinosis. A high-molecular-weight glycoprotein was isolated, purified and partially characterized from the insoluble pulmonary secretions accumulating in lungs of patients suffering from pulmonary alveolar proteinosis. On electrophoresis in 5% polyacrylamide gel in the presence of sodium dodecyl sulphate and 2-mercaptoethanol, the purified protein gave one major band as detected by Coomassie Blue as well as with periodic acid/Schiff staining. An apparent mol.wt. of 250000 was estimated for this glycoprotein. Amino acid analysis showed that it contains hydroxyproline, and relatively high amounts of glycine, glutamic acid, aspartic acid and leucine. It contains approx. 6% hexose, 3% sialic acid and 2% glucosamine. The neutral sugars are galactose, mannose and fucose. An antiserum prepared in rabbits against this high-molecular-weight glycoprotein cross-reacted with two smaller glycoproteins (mol.wts. 62000 and 36000) isolated from the same pulmonary secretions of these patients. A complementary observation was also made when this large alveolar glycoprotein cross-reacted with an antiserum prepared in rabbits against the smaller glycoprotein (mol.wt. 36000). It appears that this high-molecular-weight glycoprotein may be the precursor of the two smaller glycoproteins present in the same diseased pulmonary secretions.


Investigation of the organization of rhodopsin in the sheep photoreceptor membrane by using cross-linking reagents. The organization of rhodopsin in the photoreceptor membrane of sheep rod outer segments was investigated by using a variety of bifunctional reagents. Of the nine reagents used, seven gave oligomeric opsin species, whereas two, copper phenanthroline and dithiobisphenyl azide, failed to cross-link the protein. In general, the cross-linked species obtained showed diminishing yields from dimer to tetramer, together with some higher-molecular-weight aggregates. It is proposed that the patterns of cross-linking arise as a result of collision complexes and best describe a monomeric organization for native rhodopsin. No significant differences between the patterns obtained with dark-adapted bleached or regenerated protein states were observed. This interpretation is discussed in relation to the postulated mechanism of action of rhodopsin.


Diamine oxidase and polyamine oxidase activities in normal and transformed cells. 1. The activity of diamine oxidase (EC 1.4.3.6) in normal rat kidney cells and in normal rat kidney cells transformed by avian sarcoma virus (B77 strain) growing in tissue culture varies with the stage of growth. There is an initial stimulation of activity by 24h after seeding, followed by a steep decline during exponential growth (48-72h). Enzyme activity decreases even further as the cells reach saturation density (confluence) after 4 days in culture when the activity in normal rat kidney cells is twice as high as that in transformed cells. 2. Differences of about the same order of magnitude are observed between transformed human cells HeLa, HEp2 (a human epithelioid carcinoma) and normal human fibroblasts, in chicken cells between normal myeloblasts and leukaemic myeloblasts, and in rats between biopsy material from normal mammary tissue and 9,10-dimethylbenz[a]anthracene-induced mammary tumours. 3. Polyamine oxidase activity also varies with the growth of transformed rat kidney cells, but shows no significant variation with the growth of normal rat kidney cells between 24 and 96h after seeding. The activity in cells at confluence is from 3- to 5-fold lower in the transformed than in the normal rat kidney cells. 4. A similar 5-10-fold decrease in activity has been found in 9,10-dimethylbenz[a]anthracene-induced mammary tumours in rats and in human oesophageal tumours. 5. Possible reasons for these observations and the contribution of these two enzymes to cellular putrescine concentrations are discussed.


A comparison of the substrate and electron-donor specificities of the methane mono-oxygenases from three strains of methane-oxidizing bacteria. 1. Methane mono-oxygenase from Methylosinus trichosporium has the same broad substrate specificity as the analogous enzyme from Methylococcus capsulatus (Bath); the enzyme from Methylomonas methanica is more specific. 2. Contrary to previous reports, NAD(P)H and not ascorbate is the required electron donor for the enzyme from Methylosinus trichosporium. 3. It is concluded that these three bacteria contain similar methane mono-oxygenases.


The cellulolytic enzymes of Botryodiplodia theobromae Pat. Separation and characterization of cellulases and beta-glucosidases. 1. Filtrates from cultures of different ages of Botryodiplodia theobromae Pat. were fractionated by gel filtration, ion-exchange chromatography and polyacrylamide-gel electrophoresis. 2. Five cellulases (C1, C2, C3, C4 and C5) were found, and their molecular weights, estimated by gel filtration, were 46000-48000 (C1), 30000-35000 (C2), 15000-18000 (C3), 10000-11000 (C4) and 4800-5500 (C5). 3. Cellulase C5 was absent from old culture filtrates. 4. Cellulase C1 had little or no activity on CM-cellulose (viscometric assay), but degraded cotton flock and Whatman cellulose powder to give cellobiose only. 5. The other components (C2-C5) produced cellobiose and smaller amounts of glucose and cellotriose from cellulosic substrates and were more active in lowering the viscosity of CM-cellulose. 6. The ratio of activities assayed by viscometry and by the release of reducing sugars from CM-cellulose increased with decrease in the molecular weights of cellulases C2-C5. 7. Cellobiose inhibited the activities of the cellulases, but glucose stimulated at low concentrations although it inhibited at high concentrations. 8. A high-molecular-weight beta-glucosidase (component B1, mol.wt. 350000-380000) predominated in filtrates from young cultures, but a low-molecular-weight enzyme (B4, mol.wt. 45000-47000) predominated in older filtrates. 9. Intermediate molecular species of beta-glucosidase (B2, mol.wt. 170000-180000; B3, mol.wt. 83000-87000) were also found. 10. Cellulases C2-C5 acted in synergism with C1, particularly in the presence of beta-glucosidase.


Identification of organic phosphorus covalently bound to collagen and non-collagenous proteins of chicken-bone matrix. The presence of O-phosphoserine and O-phosphothreonine in non-collagenous proteins, and their absence from phosporylated collagen. Non-collagenous phosphoproteins, almost all of which can be extracted in EDTA at neutral pH in the presence of proteinase inhibitors, are identified in the matrix of chicken bone, and are therefore not covalently bound to collagen. Similarly, all the peptides containing gamma-carboxyglutamic acid are present in the EDTA extract and none in the insoluble residue, confirming that none is covalently linked to chicken bone collagen. However, organic phosphorus is also found to be present in chicken bone collagen, principally in the alpha2-chains. Of the total protein-bound organic phosphorus present in chicken bone matrix, approx. 80% is associated with the non-collagenous proteins and 20% with collagen. The soluble non-collagenous proteins contain both O-phosphoserine and O-phosphothreonine and these account for essentially of their organic phosphorus content. In contrast, collagen contains neither O-phosphoserine nor O-phosphothreonine. Indeed, no phosphorylated hydroxy amino acid, phosphoamidated amino acid or phosphorylated sugar could be identified in purified components of collagen, which contain approximately four to five atoms of organic phosphorus per molecule of collagen. Peptides containing organic phosphorus were isolated from partial acid hydrolysates and enzymic digests of purified collagen components, which contain an as-yet-unidentified cationic amino acid. These data, the very high concentrations of glutamic acid in the phosphorylated peptides, and the pH-stability of the organic phosphorus moiety in intact collagen chains strongly suggest that at least part of the organic phosphorus in collagen is present as phosphorylated glutamic acid. This would indicate that the two major chemically different protein fractions in chicken bone matrix that contain organic phosphorus may represent two distinct metabolic pools of organic phosphorus under separate biological control.


Identification of hydroxyapatite crystals in synovial fluid. A semiquantitative technique employing (14C) ethane-1-hydroxy 1, -1-diphosphonate (EHDP) binding has been used to detect crystals, presumably hydroxyapatite, in human synovial fluid samples which were handled to prevent the formation of artifactual mineral phase. Binding material was found in 29% of non-inflammatory and in none of inflammatory joint fluids. Nuclide binding material was strongly correlated with the presence of CPPD crystals and with radiographic evidence of cartilaginous degeneration.


Correlation of total body potassium and leukemic cell mass in patients with chronic lymphocytic leukemia. Total body leukemic mass in patients with chronic lymphocytic leukemia (CLL) was measured by quantitation of total body potassium (TBK) with a whole-body counter. In addition, the predicted normal total body potassium (Kp) for each patient was calculated from an empirically derived relationship involving height, weight, age, and sex. Both the absolute TBK and the relative excess of total body potassium (TBK/Kp) were related to the stage of disease. Patients in the early stages of CLL were found to have lower TBK and TBK Kp than patients in the late stages of disease. Both of these parameters increased with the successively advanced stages of the disease. The clinically monitored reduction of leukemic cell mass following therapy was accompanied by reductions in TBK and TBK/Kp. Data presented support the notion that TBK/Kp is a useful indicator of the total body leukemic mass. Furthermore, the results of these studies quantitatively validate the proposed clinical staging system for CLL. Quantitation of TBK by a whole-body counter is an accurate and noninvasive procedure and does not require administration of isotopes.


[Release of antigens into the culture medium by LS fibroblasts in culture]. LS fibroblasts cultivated for 1 or more days in unsupplemented Eagle's MEM release antigens into the medium, without showing any evidence of lysis, but on the contrary continuing to proliferate. These antigens give up to three precipitation lines when tested by means of immunodiffusion and immunoelectrophoresis against specific rabbit antisera; one or two of them give identity reactions with antigens obtained by repeatedly washing the fibroblasts with balanced salt solutions. Evidence has been obtained that they are surface components, easily stripped or spontaneously shed by the cells.


[Vascular thrombosis after catheterization of the umbilical artery]. Two newborns with vascular thromboses due to catheterization of the umbilical artery are reported; one of them died and in the other one it was necessary to amputate the leg. Considerations are made on the predisposing factors for this complications, as well as the indications and precautions for this technic.


Ebola virus: a comparison, at ultrastructural level, of the behaviour of the Sudan and Zaire strains in monkeys. Histopathological and electron microscopical examination of human liver specimens collected during the Ebola haemorrhagic fever outbreaks in Zaire and Sudan indicated that Zairean strains of the virus produced more extensive lesions. Experimental infection of rhesus monkeys wiht Zairean and Sudanese strains of Ebola virus produced similar changes to those found in man. In Zairean strain infections large numbers of virus particles were found in the liver, lung and spleen accompanied by extensive necrosis in the spleen. In Sudan strain infections particles were found only in the liver and in greatly reduced numbers. The main distinction lay in the high proportion of aberrant particles found with the Sudanese strain. The possibility of these being defective particles is discussed.


The pathology of untreated and antibiotic-treated experimental tularaemia in monkeys. Grivet monkeys were infected intranasally with the virulent Schu-S4 strain of F. tularensis. One group of animals remained untreated and two other groups received a 7-day course of kanamycin therapy starting on either the third or fourth day after infection. Untreated monkeys developed pyrexia and mucopurulent oculonasal discharge and died 5--7 days after infection. All had pyogranulomatous lesions in the liver, spleen, respiratory tract and lymph nodes. Electron microscopy of liver and spleen showed phagocytosis of F. tularensis organisms by macrophages and polymorphonuclear leucocytes, but many bacteria survived phagocytosis and were released on destruction of the cells. Kanamycin therapy enabled most monkeys to survive the disease, but it did not prevent the development of persistent lesions in all animals. Caseous nodules were larger and more widespread in the organs of monkeys in which treatment was delayed until the fourth day of infection.


Changes in whole blood and serum components of grivet monkeys with experimental respiratory Francisella tularensis infection. Grivet monkeys infected with virulent Francisella tularensis Strain Schu S4 showed significant early changes in serum levels of trace metals, triglycerides and activities of alkaline phosphatase, lactate dehydrogenase and alpha-hydroxybutyrate dehydrogenase. Free amino acid levels decreased slightly and there was a marked increase in the phenylalanine: tyrosine ratio. Serum lysozyme activity and seromucoid levels also increased. Kanamycin therapy produced remission of overt signs but the changes in blood constituents were less readily affected. Immunization with the live vaccine strain of F. tularensis induced transient responses similar to those resulting from Schut S4 infection. Immunized monkeys subsequently challenged with the virulent Schu S4 strain showed no clinical signs or marked changes in blood constituents.


Cross-reactivity with mouse antigens in the ferritin immunogenetic (IR-gene) system. Structural similarity between antigens and self molecules could be responsible for low antibody responses in different immunogenetic (IR-gene) systems. B10.M and B10.D2 strains are high responders, whilst A. Thy-1-1 mice are low responders, following primary immunization with ferritin in saline. Cross-reactivity between mouse-self antigens and ferritin was tested by antigen excess and radioimmunoassay techniques, using cells obtained from normal, unimmunized high- and low-responder mice, to compete for specific antibody. Low-responder A.Thy-1-1 mouse cells consistently displaced more anti-ferritin antibodies than did high-responder B10.M and B10.D2 mouse cells at varying antibody and cell concentrations and these differences were statistically significant (P less than 0.001). It is suggested that the responder status of different strains of mice, following primary immunization with ferritin in saline, could be explained by the degree of cross-activity between self determinants and antigen, such that low responders cross-react to a greater degree with the test antigen than do high-responder mice. A similar mechanism of cross-reactivity could operate in the pathogenesis of HLA-linked diseases.


The incidence of antibodies to low frequency antigens (LFA) in plasmapheresis donors with hyperimmune Rh antisera. Over 50% of plasmas from plasmapheresis donors hyperimmunized for Rh antibodies were found to contain antibodies to low frequency red cell antigens (LFA). The majority of these plasmas contained antibodies to more than one LFA. On the other hand, the incidence of these antibodies in immune plasma from control plasmapheresis donors was markedly lower. The significance of the high incidence of these antibodies in grouping reagents is discussed.


Proton irradiation of malignant melanoma of the ciliary body. This is our first case of malignant melanoma of the ciliary body treated with proton beam irradiation, a technique that we developed for irradiating choroidal melanomas. After 21 months of follow-up no growth of the tumour has been observed, and shrinkage of the tumour was noted on the follow-up photographs and by ultrasonography. The 32P uptake test, which was positive before treatment, turned negative 14 months after irradiation. The described technique of proton beam irradiation might offer an alternative for the treatment of ciliary body melanomas when the present techniques of iridocyclectomy cannot be applied because of the size of the lesion.


Stability of polysome-associated polyadenylated RNA from soybean suspension culture cells. The half-life of polysome-associated, poly(A)-RNA in exponentially growing soybean (Glycine max) suspension culture cells was determined with pulse-chase experiments. Based on a best fit from a computer analysis of the data, two decay components for poly(A)-RNA were found. One component had a half-life of approximately 0.6 h, while the other had a half-life of about 30 h, similar to the doubling time of the cultures. At the beginning of the chase period, the short-lived component represented approximately 90% of the total poly(A)-RNA in the polysomes. This percentage decreased with time so that, under steady-state conditions, the long-lived component probably represented the majority of poly(A)-RNA.


Nuclear protein modification and chromatin substructure. 3. Relationship between poly(adenosine diphosphate) ribosylation and different functional forms of chromatin. The relationship between poly(adenosine diphosphate) ribosylation of nuclear proteins and functionally different forms of chromatin from mid-S-phase HeLa nuclei was investigated. The major observations emerging from this study were that unique nonhistone proteins were modified in mid-S-phase HeLa nuclei. The major acceptor for poly(adenosine diphosphate-ribose) [poly(ADP-Rib)] was an internucleosomal nonhistone protein (protein C; 125 000 molecular weight). Histones H3, H1, H2b, and H2a but not H4 were ADP-ribosylated in S-phase nuclei. Chromatin fragments preferentially released by micrococcal nuclease were enriched in nonhistone proteins, poly(ADP)-ribosylated nuclear proteins, poly(ADP-Rib) polymerase activity and nascent DNA from the DNA replicating fork. In extended forms of chromatin, contiguous to the DNA replicating fork, poly(ADP-Rib) polymerase was maximally active. However, in chromatin distal to the replicating fork (i.e., more condensed structures), nucleosomal histones and histone H1 were not significantly ADP-ribosylated, and poly(ADP-Rib) polymerase activity was depressed two- to threefold. The data suggest that a subset of nucleosomes in extended regions of chromatin is subject to extensive ADP ribosylation.


Purification and characterization of a thyrotropin-releasing hormone deamidase from rat brain. This report describes the purification of a rat brain thyrotropin-releasing hormone (TRH) deamidating enzyme to apparent homogeneity. Criteria for purity include sodium dodecyl sulfate and disc gel electrophoresis, as well as isoelectric focusing (pI = 4.5). Enzyme purification was facilitated by development of a rapid and sensitive continuous assay using the substrate L-pyroglutamyl-Nim-benzylhistidyl-L-prolyl-beta-naphthylamide, which, upon hydrolysis of the naphthylamide, results in the appearance of the fluorescent product, beta-naphthylamine (beta NA). With this substrate the homogeneous enzyme had a specific activity of 14.5 mumol of beta NA min-1 mg-1. The only peptide product formed was shown to be L-pyroglutamyl-Nim-benzylhistidyl-L-proline. Hydrolysis of [L-prolyl-2,3-3H]TRH was shown to yield L-pyro-glutamyl-L-histidyl-L-proline as the only radiolabeled product. Characterization of the brain deamidase by gel filtration chromatography and sodium dodecyl sulfate gel electrophoresis indicated that the enzyme consists of a single polypeptide chain having molecular weights of 70,000 and 73,500, respectively. Rat brain TRH deamidase has an apparent Km of 34 micron, and a pH optimum between 7 and 8 using L-pyroglutamyl-Nim-benzylhistidyl-L-prolyl-beta-naphthylamide as a substrate. With this substrate, TRH was shown to be a competitive inhibitor with an apparent Ki of 120 +/- 20 micron.


Heterogeneity of delta-crystallins of the embryonic mallard lens. Correlation between subunit compositions and isoelectric points. delta-Crystallins from the lenses of embryonic mallards (Anas platyrhynchos) were analyzed with respect to native and subunit molecular weight, subunit composition, and isoelectric point. NaDodSO4-urea-polyacrylamide gel electrophoresis showed that unfractionated mallard delta-crystallins are composed of approximately equal amounts of subunits with molecular weights near 47 000 and 48 000. Agarose gel chromatography showed that the embryonic mallard delta-crystallins have native molecular weights slightly less than 200 000. Thus, embryonic mallard delta-crystallins appear to be tetramers. Five major and nine minor delta-crystallins were resolved by isoelectric focusing. The five predominant delta-crystallins each cross-reacted with antichick delta-crystallin antiserum, and each had a different proportion of the larger and smaller subunits, indicating a direct relationship between the isoelectric point and the subunit composition. The presence of numerous, minor species of native delta-crystallins with different isoelectric points suggested that the subunits possess charge heteogeneity as well as size heterogeneity.


Purification and preparation of antibody to RNA polymerase II stimulatory factors from Ehrlich ascites tumor cells. An improved method was developed for purification of the protein termed S-II that specifically stimulates RNA polymerase II of Ehrlich ascites tumor cells. The specific activity of the final preparation was 400 000 units/mg of protein, which is about 30-fold higher than that of the previous preparation [Sekimizu, K., et al. (1976) Biochemistry 15, 5064]. The final preparation gave a single band on both sodium dodecyl sulfate and nondenaturing gel electrophoresis, and the protein extracted from the band on nondenaturing gel had stimulatory activity. S-II is a basic protein with a molecular weight of 40 500. The fundamental characteristics of S-II determined with the previous preparation were confirmed with completely purified S-II. A specific antibody to S-II was prepared. This antibody inhibited only the stimulatory activity of S-II and did not affect the activity of RNA polymerase II itself. Thus, S-II is probably not a component of the multimeric proteins of RNA polymerase II.


Cell-free protein synthesis in lysates of Drosophila melanogaster cells. A procedure is described for preparing cell-free protein synthesizing lysates from Drosophila melanogaster tissue culture cells and embryos. Preparation of translationally active lysates from tissue culture cells is dependent on the presence of rat liver supernatant during cell lysis to inhibit ribonuclease activity. After micrococcal nuclease treatment of the lysate, protein synthesis is dependent on the addition of exogenous messenger RNA. The fidelity of translation is very high. The conditions for optimal translation have been determined. In addition, the effects on translation of a variety of supplements, including rat liver supernatant, have been analyzed. The products of translation by the Drosophila lysate have been compared with those of wheat germ extracts and of micrococcal nuclease treated rabbit reticulocyte lysates. Translation in vitro of bovine parathyroid hormone messenger RNA yielded two products tentatively identified as preproparathyroid hormone and proparathyroid hormone, as well as an unidentified third product. This result suggests that insect enzymes can accurately process mammalian precursor proteins.


A monolayer study of the reaction of trinitrobenzene sulphonic acid with amino phospholipids. The reaction of trinitrobenzene sulphonic acid with amino phospholipids, and in particular phosphatidylethanolamine has been studied by the monolayer technique. Injection of trinitrobenzene sulphonic acid under a monolayer of amino phospholipid results in an increase in surface pressure. The rate and extent of the pressure change is greatly affected by the initial surface pressure, the fatty acid composition of the lipid, and the presence of other non-reactive lipids, especially negatively charged phospholipids. The extent of the reaction was measured with 32P-labelled phospholipids isolated from Bacillus subtilis. Only about 80% of the phosphatidylethanolamine in the monolayer could be converted to its trinitrophenyl derivative. In the presence of negatively charged phospholipids such as cardiolipin or phosphatidylglycerol, a further 20% decrease in the trinitrophenylation of phosphatidylethanolamine was found. The pressure increase occurring during trinitrophenylation could also be correlated with the extent of the reaction by comparison of the force-area curves of pure phosphatidylethanolamine, its trinitrophenyl derivative and mixtures of both compounds. The data may offer an explanation for the observation that incomplete labelling of amino phospholipids frequently occurs in natural membranes and furthermore indicate that the use of chemical labelling techniques in the study of lipid asymmetry in biological membranes must be approached with great caution.


Phase changes of cardiolipin vesicles mediated by divalent cations. Small unilamellar vesicles were prepared from cardiolipin and produced the hexagonal II phase when dialyzed against CaCl2 or MgCl2. Upon removal of the cation by dialysis against EDTA large unilamellar vesicles were formed. The events of the transition from the lamellar to hexagonal phase and back to the lamellar phase are described.


The effect of protein depletion on the rate of protein synthesis in rat liver. In order to understand the mechanism of decreased protein synthesis in the liver of rats fed a protein-free diet, the average polypeptide chain assembly time (tc) was measured by the method of Mathews et al. (J. Biol. Chem. (1973) 248, 1329). For rats fed a normal diet, tc in liver in vivo was 1.28 min. A 10-day period of protein depletion led to a value of tc = 2.08 min, corresponding to a 38% depression in polypeptide elongation rate. Protein depletion caused an extensive breakdown of hepatic polysomes and refeeding of a complete mixture of amino acids resulted in rapid recovery of polysomal profile. But tc in the liver of the refed animals gave still depressed value of 1.95 min. The amount and size distribution of poly(A)-containing mRNA in the liver, as determined by [3H]poly(U) hybridization, were the same for normal and depleted groups. These results suggest that both initiation and elongation steps of protein synthesis are depressed in the liver of protein-depleted rats. Refeeding of amino acid mixture rapidly restores initiation but not elongation activity.


Rat testis lipoxygenase-like enzyme. Characterization of products from linoleic acid. The linoleate oxidation products of the affinity chromatography-purified lipoxygenase-like enzyme isolated from rat testes microsomes were characterized. Three types of reaction products separated by thin-layer chromatography were generally present: polar byproducts (A and B) and hydroperoxides. The methyl hydroxystearates obtained from the enzymically produced hydroperoxides were analysed by gas-liquid chromatography and showed a ratio of 67% 13-hydroxy isomer to 33% 9-hydroxy isomer. The major polar byproduct was analysed by infrared spectra, nuclear magnetic resonance and mass spectrometry (of the toluene-p-sulphonyl derivative) and its structure was established as 13-hydroxy-12-oxo-octadec-cis-9-enoic acid. The possibility of the existence of a linoleate hydroperoxide isomerase in the affinity-purified preparation is discussed.


Mutant and immunochemical studies on the involvement of cytochrome b5 in fatty acid desaturation by yeast microsomes. The involvement of cytochrome b5 in palmitoyl-CoA desaturation by yeast microsomes was studied by using yeast mutants requiring unsaturated fatty acids and an antibody to yeast cytochrome b5. The mutants used were an unsaturated fatty acid auxotroph (strain E5) and a pleiotropic mutant (strain Ole 3) which requires either Tween 80 and ergosterol or delta-aminolevulinic acid for growth. Microsomes from the wild-type strain possessed both the desaturase activity and cytochrome b5, whereas those from mutant E5 contained the cytochrome but lacked the desaturase activity. Microsomes from mutant Ole 3 grown with Tween 80 plus ergosterol were devoid of both the desaturase activity and cytochrome b5, but those from delta-aminolevulinic acid-grown mutant Ole 3 contained cytochrome b5 and catalyzed the desaturation. The cytochrome b5 content in microsomes from mutant Ole 3 could be varied by changing the delta-aminolevulinic acid concentration in the growth medium, and the desaturase activity of the microsomes increased as their cytochrome b5 content was increased. The antibody to yeast cytochrome b5, but not the control gamma-globulin fraction, inhibited the NADH-cytochrome c reductase and NADH-dependent desaturase activities of the wild-type microsomes. It is concluded that cytochrome b5 is actually involved in the desaturase system of yeast microsomes. The lack of desaturase activity in mutant Ole 3 grown with Tween 80 plus ergosterol seems to be due to the absence of cytochrome b5 in microsomes, whereas the genetic lesion in mutant E5 appears to be located at ther terminal desaturase.


Temperature-induced changes in fatty acid unsaturation of Tetrahymena membranes do not require induced fatty acid desaturase synthesis. The usual rise in phospholipid-bound palmitoleic acid of Tetrahymena pyriformis cells slowly acclimating to low temperature exposure can be prevented by cycloheximide. This reduction in fatty acid desaturation is not caused by specific inhibition of a temperature-induced synthesis of a fatty acid desaturase but rather by a general effect equally conspicuous in isothermal cells. Cycloheximide-inhibited cells chilled and analyzed quickly, before long term ill effects of the drug are expressed, exhibit the rise in unsaturated fatty acids typical of temperature-acclimating cells.


[Multiple molecular forms of erythrocyte carbonic anhydrase of sheep (author's transl)]. Three major forms CI, CII and CX and the minor component CI1v of ovine erythrocyte carbonic anhydrase have been identified in the ethanol chloroform extract of the hemolysate by electrophoresis on cellulose acetate and isoelectrofocusing. Isolation of the four forms was achieved by chromatography on DEAE-Sephadex A-50. Comparative studies suggest that: (1) form CI1v is a modified form of CI, (2) form CX is CI plus glutathione and (3) forms CI and CII are isoenzymes which differ in their primary structure by one Lys leads to Thr substitution 35 residues from the N-acetylated terminus.


Purification and characterization of the respiratory nitrate reductase of Bacillus licheniformis. 1. Respiratory nitrate reductase of Bacillus licheniformis was extracted from the bacterial membranes by treatment with deoxycholate and purified to a homogeneous state by means of gel chromatography and anion-exchange chromatography. 2. The enzyme (Mr = 193,000, s20, w = 8.6) consists of two subunits, having apparent molecular weight of 150,000 (alpha subunit) and 57,000 (beta subunit), which are present in an equimolar ratio. It does not contain carbohydrate. Ageing of the enzyme appears to result in splitting of the polypeptide chains at specific sites followed by dissociation and reassociation of the digestion products in various combinations. 3. In contrast to Klebsiella aerogenes repiratory nitrate reductase, which is isolated in a tetrameric form that can be reversibly dissociated into a monomeric form by detergents, B. licheniformis nitrate reductase, after isolation, is always present in a monomeric form. This property is related to the difference in membrane localization of the enzyme in the two organisms. 4. B licheniformis nitrate reductase contains 6.9 atoms of non-heme iron, 6.7 atoms of acid-labile sulfide and 0.93 atoms of molybdenum per molecule of enzyme. The molybdenum seems to be part of a low-molecular weight peptide Mo-cofactor) to which it may be bound by interaction with thiol-groups. 5. Antiserum against the native enzyme contains antibodies against both subunits as well as the Mo-cofactor. The Mo-cofactor does not have any antigenic determininants in common with either the alpha or the beta subunit. Also neither subunit cross-reacts with antiserum against the other subunit. Whereas the respiratory nitrate reductases from K. aerogenes and Escherichia coli are immunologically related, the native enzyme from B. licheniformis does not show any cross-reaction with antiserum prepared against either the K. aerogenes or the E. coli enzyme.


Subunit structure of the mouse epidermal keratin filament. The two proteins which are the subunits of mouse epidermal keratin filaments have been isolated from fully differentiated epidermis (stratum corneum), viable differentiating cells and cells grown in culture. The proteins have molecular weights of 68 000 and 60 000, consist of families of very similar species, have common N-terminal (N-acetylserine) and C-terminal (glycine) residues, contain 35--40% alpha-helix and are immunologically cross-reacting. In mixtures, the two proteins polymerize in vitro into native-type keratin filaments that are 70--80 angstrom in diameter, up to 30 micrograms long, possess a characteristic alpha-type X-ray diffraction pattern and contain the subunits in the precise molar ratio of 1 : 2 or 2 : 1.


Artifacts in isoelectric focusing of the microsomal enzymes cytochrome P-450 and NADPH-cytochrome P-450 reductase. Highly-purified rat liver microsomal cytochrome P-450 and NADPH-cytochrome P-450 reductase (NADPH-ferricytochrome oxidoreductase, EC 1.6.2.4) preparations gave rise to a large number of bands under a variety of isoelectric focusing conditions, as observed after staining for either zymogen or protein. The binding patterns were not independent of sample concentration and position of application, and eluted bands did not refocus as expected. The artifactual heterogeneity is attributed to strong protein-protein interactions and perhaps to complexation of proteins with carrier ampholytes. These findings suggest caution in using isoelectric focusing to resolve mixtures of membrane proteins.


Phenolic and nonphenolic ring deiodinations of iodothyronines in cultured hepatocarcinoma cell homogenate from monkey. Iodothyronine monodeiodinase activities in homogenates of cultured monkey hepatocarcinoma cells were measured by the deiodination of [3.5-(125)I]-diiodo-L-thyronine or 3-[3',5'-(125)I]triiodo-L-thyronine (phenolic ring-labeled 'reverse' triiodothyronine). The assay system utilized a small ion-exchange column (AG50W-X4, O.9 X approximately 1 cm) to measure 125I-. Both deiodinases were destroyed by boiling for 1 min. Maximal nonphenolic ring deiodination was observed at pH 7.9 whereas maximal phenolic ring deiodination was at pH 6.3. Both reactions were enhanced strongly by dithiothreitol (0.1-5mM), and slightly by 5 mM beta-mercaptoethanol. Phenolic ring deiodination was strongly inhibited by 0.1 mM propylthiouracil. Nonphenolic ring deiodination was accelerated by EDTA (1.2 MM) and inhibited by Mg(2+) (5mM). Methylmercaptoimidazol and Mg(2+), Ca(2+) and Mn(2+) (0.1-1.0 mM) had little or no effect on either reaction, but Zn(2+) (0.1 mM) strongly inhibited both. Both reactions were inhibited by excess iodothyronine analogues at 10 mM to 10 micron M, and thyroxine was shown to be a competitive inhibitor in both cases. On the basis of relative affinities and inhibitory effects, it appears that the order of affinity for the phenolic ring deiodinase is 3,3',5'-triiodo-L-thyronine(rT3) greater than L-thyroxine(T4) greater than 3,5,3'-triiodo-L-thyronine(T3), whereas for the nonphenolic ring deiodinase the order is T3 greater than T4 greater than rT3. Diiodotyrosine did not affect their deiodination.


Interaction of phenosafranine with nucleic acids and model polyphosphates. III. Heterogeneity in phenosafranine interactions with DNA base pairs. Fluorescence and circular dichroism spectral measurements, thermal denaturation studies and binding competition experiments with netropsin and actinomycin D were carried out in systems containing phenosafranine bound to DNA's differing in base composition. The investigated properties exhibit a heterogeneity related to the content of A.T and G.C pairs in DNA and to the nature of phenosafranine binding modes. At low level of saturation of binding sites (r less than 0.1) phenosafranine does not show strong preference for any of the DNA base pairs in the overall binding. However, the strong monomer non-cooperative binding outside the helix (mode I1) occurs predominantly, even though not exclusively in G.C rich regions. The strong binding modes involving intercalated dye molecules (mode I2 and eventually mode II1) prevail in A.T rich regions. These binding modes become the principal types of strong phenosafranine interaction with DNA when the level of saturation of binding sites increases, i.e. at r greater than 0.1.20


Metabolism of diethylstilbestrol: identification of a catechol derived from dienestrol. The enzymatic oxidation of E-3,4-bis-(p-hydroxyphenyl)-hex-3-ene (diethylstilbestrol) by either mushroom tyrosinase or rat liver microsomes in the presence of NADPH and air yields a catechol. Upon further oxidation of both compounds with periodate and condensation of the resulting o-quinones with o-phenylenediamine, phenazines are produced. The phenazines derived from the products of both the plant and animal enzyme systems are identical to the product obtained by oxidation of diethylstilbestrol with potassium nitrosodisulfonate and condensation of the o-quinone produced with o-phenylenediamine. High and low resolution mass spectra of the phenazine are consistent with its derivation from a catechol having two fewer hydrogens than diethylstilbestrol.


Structural determination of 'cord factor' from a Corynebacterium diphtheriae strain by a combination of mass spectral ionization methods: field desorption cesium cationization and electron impact mass spectrometry studies. The composition and structure of a preparation of 'cord factor' (di-beta-hydroxy acyl trehaloses) from Corynebacterium diphtheriae have been determined by a combination mass spectral ionization methods. The methods were tested by means of synthetic 6,6'-dicorynomycolate of alpha-D-trehalose prior to their use on natural products. The determination of the molecular weight of the components and the estimation of their relative abundance in the natural mixture were made by field desorption mass spectrometry and cationization by means of cesium iodide. At least 22 molecular species differing from the chain length and the unsaturation degree were detected (carbon numbers C76 to C66). The position of acylation and the nature of the acyl chains were obtained from the electron impact mass spectra of the trimethylsilyl derivatives. The trehalose molecular was found to be esterified by a complex mixture of corynomycolic acids (3-hydroxy 2-alkyl fatty acids) which were present as saturated, mono-unsaturated and di-unsaturated homologues (carbon numbers C32 to C24). The 6 and 6' sites of acylation were the only detectable ones.


Studies on the mechanism of enzymatic hydrolysis of cellulosic substances. Most cellulosic substances contain appreciable amounts of cellulose and hemicellulose, which on enzymatic hydrolysis mainly yield a mixture of glucose, cellobiose, and xylose. In this paper, studies on the mechanisms of hydrolysis of bagasse (a complex native cellulosic waste left after extraction of juice from cane sugar) by the cellulase enzyme components are described in light of their adsorption characteristics. Simultaneous adsorption of exo- and endoglucanases on hydrolyzable cellulosics is the causative factor of the hydrolysis that follows immediately after. It supports the postulate of synergistic enzyme action proposed by Eriksson. Xylanase pretreatment enhanced the hydrolysis of bagasse owing to the creation of more accessible cellulosic regions that are readily acted upon by exo- and endoglucanases. The synergistic action of the purified exoglucanase, endoglucanase, and xylanse has been found to be most effective for hydrolysis of bagasse but not for pure cellulose. Significant quantities of glucose are produced in beta-glucosidase-free cellulase action on bagasse. Individual and combined action of the purified cellulase components on hydrolysis of native and delignified bagasse are discussed in respect to the release of sugars in the hydrolysate.


Affinity chromatography of endoglucanase of Trichoderma viride by concanavalin A-agarose. Endoglucanase (C kappa cellulase) and cellobiase are often cross-contaminated in separation procedures by ion-exchange chromatography such as DEAE-cellulose. By using concanavalian A (Con A)-agarose chromatography, C kappa cellulase and cellobiase from Trichoderma virde can be separated. C kappa cellulase showed affinity toward Con A. indicating a glycoprotein containing alpha-D-mannopyransyl and alpha-D-glucopyranosyl end groups or internal 2-O-D-mannopyranosyl residues in sugar moieties. This method provides a way to estimate the quantities of C kappa enzyme produced by T. viride and possibly by other organisms.


Penicillin G production by immobilized whole cells of Penicillium chrysogenum. Penicillium chrysogenum was immobilized in polyacrylamide gel prepared from 5% acrylamide monomers (85% acrylamide and 15% N,N'-methylene bisacrylamide). Penicillin produced from glucose by the immobilized mycelium was 17% of that produced by washed mycelium. However, the activity of penicillin production of the washed mycelium decreased with repeated use. On the other hand, the activity of the immobilized mycelium increased initially and decreased gradually with repeated use. The rate of oxygen uptake of the immobilized mycelium was about 30% of that of the washed mycelium. The immobilized mycelium required oxygen for the production of penicillin.


Association of beta-glucosidase with intact cells of Thermoactinomyces. The location of the B-glucosidase activity in a whole culture broth of the thermophilic organism Thermoactinomyces has been studied. Little beta-glucosidase activity was found in the culture filtrate, while the culture solids contained the major part of the activity of the whole culture broth. The activity does not appear to be adsorbed to the culture solids; rather there is evidence that it is an intracellular soluble enzyme(s). The pH and temperature optima for a crude beta-glucosidase preparation were determined to be pH 6.5 and 50--55 degrees C. Enzyme activity studies indicate that the same enzyme(s) accounts for the beta-glucosidase and the cellobiase activities. The validity of using the filter paper activity of culture filtrates from Thermoactinomyces to predict the total saccharification of cellulosic materials to glucose is discussed.


Improved nonporous magnetic supports for immobilized enzymes. Ni powders coated by deposition of TiO2 or controlled oxidation to NiO develop substantial resistance to corrosion. Chymotrypsin immobilized to these coated Ni supports shows very high stability of activity on storage. Chymotrypsin immobilized by adsorption and glutaraldehyde crosslinking was fairly rapidly eluted under operational conditions in the presence of substrate. If 3-aminopropyltriethoxysilane (APS) was used to produce a covalent linkage, desorption of enzyme still occurred because of relatively unstable bonding of the silane to the oxide surface. A more stable attachment was produced by joining together many silane links with a layer of polyglutaraldehyde. The mechanism of action of APS as a coupling agent under these conditions is discussed. gamma-Fe2O3, and particularly a Mn-Zn ferrite, are suitable magnetic support materials available with smaller particle sizes. Particles below 1 mum give the expected higher specific activities of immobilized enzymes.


Light-induced enzyme system leading to pigmentation. Alpha-Chymotrypsin was light sensitized by acylating with cis-cinnamoyl ester, a substrate interconvertible to the trans form by ultraviolet (UV) light. The degree of acylation by this method was complete leaving no residual activity of the enzyme. Upon UV irradiation the inhibited enzyme regained about 70% of its original activity, thereby adding light-sensitiveness to the proteolytic enzyme. In seeking a photographic application of the light-sensitized enzyme, a pigmenting enzyme was incorporated with it. The coupled enzyme system was shown to exhibit a light signal in the form of dark pigment slurry.


Immobilization of enzymes on activated carbon: selection and preparation of the carbon support. Based upon its superior catalytic activity for H2O2 decomposition, a bituminous coal-based activated carbon was selected for investigations of pretreatment and enzyme immobilization methods. Pretreatments considered include acid washing, exposure to strong oxidizing agents, contact with concentrated peroxide solutions, nitration and amination, isothiocyanate derivatization, silanization, and stearic acid coating. Effects of these pretreatments on morphology and trace-metal content of the carbon pellets have been studied using scanning electron microscopy and dispersive analysis of x rays. Immobilization of glucoamylase by adsorption, glutaraldehyde crosslinking, and covalent attachment to carbon activated by water-soluble diimide or diazotization have been examined. These different enzyme-carbon catalysts have been characterized by their enzyme loading, enzyme activity, catalytic activity for H2O2 decomposition, or combinations of these measures of performance.


[Modeling of determinant and dependent foci of epileptic activity in the rat cerebral cortex]. Foci of increased excitability were created in acute experiments on rats by means of weak strychnine solutions working at independent regimens. The hyperactive excitability focus induced by means of concentrated strychnine solutions played the role of a determinant structure. The importance of the latter is in the fact that it determines the activity character of other epileptogenic foci, enhances their convulsive activity, unites them into a single functional complex and determines the behaviour of the complex as a whole. This complex can be destroyed by depression of the determinant focus activity. Switching off any dependent foci included into this complex fails to destroy that latter. Results of the investigations confirmed on the new model the general concept of the role played by the determinant structures in the brain activity.


[Intensification of antibacterial and antiviral nonsusceptibility and of the immune response with an officinal RNA preparation]. Sodium nucleinate increased essentially the insusceptibility of mice to pathogenic escherichia, strain O26, Pr. vulgaris, Ps. aeruginosa, Ser. marcescens, and produced a total stimulating effect on the nonspecific bacterial resistance; analogous stimulating activity was found in the homologous low polymeric RNA from the liver. Sodium nucleinate intensified the insusceptibility of the animals to the tick-born encephalitis and encephalomyelitis viruses, and increased the antibody-forming cells count. The side-effect of heat-inactivated vaccine from pathogenic escherichia was reduced in animals inoculated with sodium nucleinate preliminarily.


[Role of vitamin A in chemical carcinogenesis in the mammary gland]. The effect of feeding rats with high doses of vitamin A on the distribution of polycyclic hydrocarbon, 7,12-dimethylbenzanthracene (DMBA) and its metabolites in different organs and blood, and also on the metabolic rate in the liver was studied following intravenous injection of the carcinogen. Hypervitaminosis A resulted in a significant decrease of the level of DMBA and its metabolites in all the organs investigated and in the blood. The rate of DMBA metabolism in the liver of the animals rose with the increase of the vitamin A dose.


Selenium 75 contamination in iodine 131 solutions. During investigations on athyreotic patients given large test doses of 131I, whole-body counting has shown the presence of 75Se contamination in some patients. The investigations were carried out to measure the small long-term component of 131I retention in such patients. In an earlier paper (Smith and Edmonds, 1977), we concluded that the effects of 75Se contamination were negligible in the patients described. Subsequent studies on a further small group of similar patients, however, have indicated that the levels of contamination are variable, and this report is intended as a warning of the possibility of measurement error due to this contamination in metabolic studies similar to those to be described. The effects of 75Se contamination observed in two out of five patients studied have been assessed.


Gentamicin- and silver-resistant pseudomonas in a burns unit. In 1977-8 gentamicin-resistant strains of Pseudomonas aeruginosa became very common in a burns unit, over 90% being resistant at the peak of the outbreak. Some strains were also resistant to silver nitrate, though silver resistance was not found in any other strains of Ps aeruginosa isolated. Unlike the gentamicin resistance, the silver resistance was unstable, and strains became sensitive on repeated subculture. All the gentamicin-resistant strains of Ps aeruginosa were of the same serotype (O:11, H:2,5). Though gentamicin resistance could be transferred in vitro from resistant strains of Ps aeruginosa to one sensitive strain of Ps aeruginosa, there was no evidence of in-vivo transfer of gentamicin resistance between strains of pseudomonas in the patients' burns, nor was there evidence of transfer of gentamicin resistance between Ps aeruginosa and enterobacteria. Carbenicillin-resistant and gentamicin-resistant Ps aeruginosa were sometimes found in the same burns, but no gentamicin-carbenicillin (doubly) resistant strains were found among the 986 strains tested during the outbreak. The outbreak of gentamicin-resistant Ps aeruginosa from burns was not reduced by stopping treatment with gentamicin and its analogues but only by segregating all patients with Ps aeruginosa in one of the two wards of the unit and admitting new patients only to the other ward.


An assessment of the Microcult-GC culture test. Microcult-GC, a miniaturised culture test, was compared with an established selective culture method for laboratory confirmation of gonorrhoea. Microcult-GC correlated 94% with the control method in identifying gonococci in male urethral specimens. Microcult-GC testing of 130 gonococci-negative vaginal swabs gave only 4.6% false-positive oxidase results, which could probably have been readily refuted by Microcult Gram stain. The test may be difficult to interpret for rectal specimens and is not appropriate for pharyngeal cultures.


Pharyngeal colonisation by Neisseria gonorrhoeae and Neisseria meningitidis in black and white patients attending a venereal disease clinic. Pharyngeal colonisation by Neisseria gonorrhoeae and Neisseria meningitidis was studies in 2000 patients attending a venereal disease clinic. Of these patients, 64% were white and 36% were black. The incidence of gonococcal infections was highest in the period from June to August. The incidence of genital or rectal infections or both was higher in the black patients. Pharyngeal colonisation by gonococci was present in 1.3% of the patients. There was no significant associations between pharyngeal colonisation and the pharyngeal symptoms, race, sex, or marital state of the patients. Pharyngeal colonisation was more frequent in patients with gonococcal infections at other sites. However, in 40.7% of the patients with pharyngeal colonisation, the pharynx was the only culture-positive site. There was no significant difference in the auxotypes or in the antibiotic susceptibility of the pharyngeal and the rectal-genital isolates except in the susceptibility to spectinomycin. Our findings do not indicate that gonococci isolated from the pharynx differ significantly from gonococci isolated from rectal or genital sites. It was notable that meningococcal colonisation of the pharynx was significantly more frequent in the white patients. This may be a genetically determined phenomenon.


Inhibition of Neisseria gonorrhoeae by normal human saliva. Saliva was found to be a powerful and specific inhibitor of Neisseria gonorrhoeae. Although 28 other species of bacteria were tested, including Neisseria meningitidis, Neisseria pharyngis var flava, Neisseria lactamica, and Neisseria catarrhalis, we failed to find any others sensitive to saliva under similar conditions. The physical properties of the inhibitory substance indicated that it might be salivary alpha-amylase. To test this hypothesis alpha-amylase was extracted from saliva and was shown to have a high antigonococcal activity. Hog pancreas alpha-amylase  also showed strong antigonococcal activity, thus the observations indicate that for the strains we tested alpha-amylase is inhibitory to gonococci. This observation indicates that either the gonococcal outer cell wall contains some unique lipopolysaccharides or that the gonococcus is unusually dependent on the integrity of these moieties. Whichever speculation proves to be true it indicates a need for a careful study of the gonococcal cell wall.


Neurophysiological aspects of pattern-sensitive epilepsy. The capacity of striped patterns (square-wave gratings) to induce paroxysmal EEG activity in a group of pattern-sensitive epileptic patients is shown to depend on: 1. The spatial frequency of the pattern. The optimum spatial frequency lies between 1 and 4 cycles/degree in every patient tested. 2. The orientation of the pattern. Although, for the patient group as a whole, no one orientation is consistently more likely to induce paroxysmal activity than any other, the responses of individual patients can show marked orientation selectivity. 3. The brightness contrast of the pattern. The probability of paroxysmal EEG activity increases dramatically as contrast is increased from 0.2 to 0.4. Red/green gratings at isoluminance fail to induce paroxysmal activity. 4. The size of the pattern. There is considerable variation between patients in the subtense of a centrally-fixated circular pattern necessary to induce paroxysmal activity with a given probability. However, for every patient an increase in the probability of paroxysmal activity from near zero to near unity is effected by an increase in the angular subtense of the pattern by a factor of two. Patterns other than square-wave gratings are capable of inducing paroxysmal activity but, in general, patterns that stimulate more than one orientation system are less epileptogenic. The above findings are compatible with the hypothesis of a seizure trigger in the striate cortex and are incompatible with a trigger confined to the lateral geniculate nucleus of the thalamus. Further evidence against a geniculate trigger is obtained from an investigation of the response of a pattern-sensitive patient to a diffuse (unpatterned) flickering field in which it is shown that the effects of counter-phase interocular flicker implicate binocular mechanisms.


Proximal limb movements in response to microstimulation of primate dentate and interpositus nuclei mediated by brain-stem structures. The cerebellar dentate and interpositus nuclei and the area of their efferent fibres have been stimulated in Cebus monkeys, using a movable microcathode. Responses consisted of eye and face movements and a stereotyped flexion of proximal parts of extremities. Very few distal limb movements were seen. The activation of proximal muscles was studied most closely. It consisted of the limited number of 5 movements: arm flexion and shoulder elevation in the forelimb and hip flexion, knee flexion and dorsiflexion of the ankle in the hindlimb. With currents of up to 100 microamperemeter these movements were elicited more readily from the interpositus and the area of efferent fibres of both nuclei as compared to the dentate nucleus. Responses were more often seen in forelimb than in hindlimb muscles, without apparent somatotopy in either nucleus. Combined forelimb-hindlimb movements were elicited from 42 per cent of effective points. Lesions placed at various locations of cerebellar output pathways demonstrated that the responses were mediated by the descending branch of brachium conjunctivum and did not require the activation of structures anterior to and including the red nucleus. The responses are interpreted to represent adjustments in flexor posture that may serve to modify maintained antigravity tonus during the initiation of volitional movements. This function, mediated by brain-stem structures, is considered to be closely associated with the activity of the lateral and intermediate cerebellum during initiation and conduction of volitional movements, which is mediated mainly through the cerebral cortex. It is stressed that control over both flexor posture and discrete distal movements is inherent in the initiation of voluntary movements.


Invariants in configurational prey selection by Bufo bufo (L.). Toads (Bufo bufo) are able to distinguish small stripes with axis orientation in the direction of motion (worm-like stripes) from those with axis orientation perpendicular to the direction of motion (anti-worm-like stripes). The configurational discrimination ability of the toad is, thus far investigated, largely invariant for (1) movement direction in the x-y plane, (2) direction of contrast--stripe against background--and (3) velocity of stripe motion.


TRH-like immunoreactivity in rat pancreas and eye, bovine and sheep pineals, and human placenta: non-identify with synthetic Pyroglu-His-Pro-NH2 (TRH). Extracts from albino rat eyes and pancreas, bovine and sheep pineals and human placenta containing TRH-like immunoreactivity were chromatographed on silica-gel plates. Comparison of elution profiles for TRH-like immunoreactivity with that of TRH revealed the presence of substances other than TRH in these samples. Chromatography of TRH-like immunoreactivity obtained from Rana pipiens skin eluted in two solvent systems produced elution profiles identical with that of synthetic Pyroglu-His-Pro-NH2 consistent with reports that frog skin contains large quantities of TRH. Implications of these findings are discussed.


Intracellular synaptic potentials of primate motor cortex neurons during voluntary movement. An intracellular recording technique was applied to the precentral motor cortex of the unanesthetized, chronically behaving monkey. Postsynaptic potentials, responsible for an initiation of the voluntary movement, were recorded. In total, 22 pyramidal tract neurons (PTNs) and 40 non-pyramidal tract neurons (non-PTNs) were successfully penetrated in 5 monkeys while the monkey was performing a flexion-extension wrist movement after a visual cue (reaction time, 200--350 msec). The neurons showed a negative membrane potential shift of at least 30 mV for more than 30 sec. A slowly rising PSP appeared 80--180 msec after the visual cue, and was 70--180 msec prior to an onset of the movement. Spike activities were superimposed upon this slow PSP with 20--80 msec rise time and 2--6 mV depolarization (8 PTNs and 6 non-PTNs). Since these depolarizations were variable in magnitude and latency, these were considered to be summated potentials of small EPSPs and hidden IPSPs. Membrane resistance was measured from an IR drop by a hyperpolarizing current (1.2 X 10(-9) A) passed through a recording electrode, and was 3.5 +/- 1.7 Momega in 18 PTNs and 4.5 +/- 2.5 Momega in 28 non-PTNs. There was a linear relationship in PTNs between membrane resistance and antidromic latency from the pontine pyramid. Because of the time course of PSPs, their possible dendritic origins were discussed.


The latency of activation of neurones in the lateral hypothalamus and substantia innominata during feeding in the monkey. To investigate whether the responses of neurones in the lateral hypothalamus and substantia innominata associated with the sight of food could control the responses of the hungry monkey to the food, the latency of activation of these neurones by food was measured. It was found that when an electromagnetically operated wide-aperture shutter opened to reveal food or non-food objects, these hypothalamic neurones responded with a latency of 150--200 msec to the food objects, and did not respond to the non-food objects. To measure the latency of the monkey's responses to the food, a visual discrimination task was set in which the monkey could lick a tube to obtain fruit juice if a food-realted visual stimulus was shown, but obtained hypertonic saline, which was aversive, if a different visual stimulus was shown. In this situation the typical latencies of the neuronal responses to the food were 150--200 msec, of the lick responses 350--500 msec, and of the EMG activity associated with these lick responses 250--400 msec. Thus the responses of these hypothalamic neurones precede the monkey's responses to the food, and could mediate the feeding and other responses of the animal to the food. It was also shown that a different population of hypothalamic neurones with responses associated with the sight of aversive visual stimuli had response latencies of 150--200 msec.


Subcellular distribution of brain peptidases degrading luteinizing hormone releasing hormone (LHRH) and thyrotropin releasing hormone (TRH). The luteinizing hormone and thyrotropin-releasing hormones have been shown to be mostly concentrated in the nerve endings of the median eminence. In contrast, the peptidases responsible for their degradation present an ubiquitous localization and most of the reports have dealt with the total soluble activity. A detailed study of the subcellular distribution of these enzymes was thus performed in cerebral cortical and hypothalamic preparations of rat brain. The activity of a soluble marker, lactic dehydrogenase, was also measured to control for possible contaminants. The results showed that only 10% of peptidase activity was present in the nerve ending preparation. Evidence is provided for a non-neglible membrane-bound enzymatic component responsible for TRH degradation at the synaptosomal level of both cortex and hypothalamus.


Brain metabolism and the acquisition of new behaviors. III. Evidence for secretion of two proteins into the brain extracellular fluid after training. Immunochemical and double labeling experiments were used to demonstrate that at least two out of three brain cytoplasmic proteins, whose metabolism is markedly influenced by behavior, are products which are secreted into the extracellular fluid (ECF) of goldfish brain. Even after 1 h of labeling, the extracts of goldfish brains with 0.32 M sucrose were found to contain highly labeled proteins. Electrophoretic analyses of the proteins, on SDS polyacrylamide gels, indicated that an increased incorporation of [3H]valine occurs for trained animals as compared to untrained controls ([14C]valine) at specific protein bands migrating at 32,000 and 26,000 daltons. The proteins in the ECF gave identical precipitin bands to goldfish brain proteins whose metabolism was responsive to the acquisition of new patterns of behavior and to proteins in CSF. These results are consistant with the hypothesis that the cells which contain the proteins and whose location in the ependymal zone was determined by immunohistochemistry can secrete the products into ECF. It is therefore quite possible that the functional sites of the proteins are away from the locus of their synthesis.


Isolation of a human B lymphocyte membrane protein with Ia-like properties. A rapid method for isolation of a major surface membrane glycoprotein from whole, unfractionated cultured human B lymphoblasts is described. After detergent solubilization the method uses gel filtration followed by affinity chromatography on Sepharose Con A and then alkaline acrylamide gel electrophoresis. Specific high-titre, rabbit antisera to the isolated protein reacted with cultured and normal peripheral blood B lymphocytes, as well as peritoneal macrophages from a renal dialysis patient. The antisera selectively inhibited the mixed lymphocyte reaction at high dilution. The protein reacted with a heterologous antiserum to HL-B antigens and contained subunits of MW 33 000 and 27 000. Resolution of the subunits, however, required a discontinuous SDS gel system. These properties indicate its similarity to murine Ia antigens. The protein was not associated with beta 2 microglobulin and showed no structural or antigenic similarity to the major erythocyte glycoprotein, glycophorin. Antisera to the protein failed to precipitate surface-radiodinated components from similarily treated extracts of cultured human T lymphoblasts. This method now makes available a reference membrane glycoprotein from a differentiated, nucleated human cell in sufficient purity and quantity for kinetic and biosynthetic studies.


Identification of lipoprotein X-like particles in rat plasma following Intralipid infusion. Fasting rats were infused with 10% Intralipid for 24 h (0.33 mL/h per 100 g body weight) and the plasma lipoproteins isolated and compared with those of fed animals and animals with bile duct ligatures as controls. There was a 6- to 10-fold increase in the free cholesterol and phospholipid content of total plasma in animals infused with Intralipid or with ligated bile ducts. The changes were largely restricted to the low density lipoproteins (d=1.019--1.063 g/mL) where free cholesterol and phospholipid increased 30- to 60-fold compared with fed control animals. Hydroxylapatite chromatography of the low density lipoprotein fractions of both Intralipid-infused and bile duct ligated animals yielded a subfraction which was rich in free cholesterol (27%), phosphatidylcholine (66%), and protein (6%); the latter was composed primarily of albumin and apo C proteins. The electrophoretic mobility and polyanionic precipitation properties of the abnormal lipoprotein were indistinguishable from those of lipoprotein X isolated from the animals with bile duct ligatures. The albumin in the abnormal lipoprotein from both groups of experimental animals was detected immunochemically only after delipidation of the lipoprotein. Twice as much of the lipoprotein X accumulated in Intralipid-infused than in the bile duct ligated animals. On rechromatography of the residual low density lipoprotein other subfractions could be isolated which possessed lipid and protein proportions intermediate between those of the lipoprotein X and of normal rat plasma low density lipoprotein. The activity of lecithin cholesterol acyl transferase was increased twofold in the Intralipid-infused animals when compared with control animals, but it decreased by 50% in the animals with bile duct ligatures. It is concluded that the unusual lipoprotein X accumulates in the plasma of Intralipid-infused animals owing to incomplete clearance of the exogenous phospholipid, which mobilized tissue cholesterol and in the form of vesicular particles serves as a lipid phase for apo C proteins. A comparable mechanism is suggested for the formation of lipoprotein X in the animals with bile duct ligature.


Effect of prolonged intermittent thyrotropin-releasing hormone administration to fetal and neonatal rats. Fetal and neonatal rats received daily subcutaneous injections of 10 microgram thyrotropin-releasing hormone (TRH) until 7 or 14 days postnatally. At 70 days the pups were challenged with 1 microgram TRH intravenously via an indwelling jugular cannula. Basal serum thyroxine, triiodothyronine, and thyroid-stimulating hormone (TSH) concentrations did not differ among the three groups. The mean TSH responses as determined by the mean peak TSH concentration and the total TSH response as determined by planimetry were not significantly different, and there was no significant difference in pituitary TSH content following the TRH challenge among the three groups. This study suggests that the integrity of the hypothalamo--pituitary axis in adult rats cannot be affected by the repeated administration of pharmacologic doses of TRH during the perinatal period.


Inhibition of passive lung anaphylaxis in the rat by AY-25,674. The antiallergic properties of AY-25,674 were studied in a model of passive lung anaphylaxis. AY-25,674 caused a dose-related inhibition of antigen-induced bronchoconstriction in the passively sensitized rat. AY-25,674 was found to be twice as potent as disodium cromoglycate (DSCG) when injected intravenously. The oral administration of AY-25,674 also effectively inhibited antigen-induced bronchoconstriction. A tachyphylactic effect of repeated administration was found when the rats were pretreated with a large dose of AY-25,674. The compound did not significantly antagonize the bronchoconstriction produced by 5-hydroxytryptamine. These results indicate that AY-25,674 is an orally effective antianaphylactic compound with a mode of action similar to that of DSCG in preventing allergic mediator release.


The effect of chemical fixatives on cell walls of Bacillus subtilis. Cell walls of Bacillus subtilis were treated with several chemical fixatives which are commonly used preparatory to electron microscopy; i.e., osmium tetroxide, formaldehyde, acrolein, crotonaldehyde, and glutaraldehyde. Dimensional analysis was performed on thin sections of fixed walls from plastic embeddings and, by means of the statistical technique of multiple comparisons, significant differences were found between wall thicknesses from the various fixations. These differences varied with the fixation time and the type of fixative used in the reaction. When compared to embedded walls which had been stained before fixation, the overall effect was a reduction in wall thickness which was attributed to fixative action and not to the embedding or staining processes. The reduction of wall thickness was even more apparent when dimensions of fixed walls were compared to published dimensions of both frozen sections and freeze-etch profiles. Since these fixatives bind to reactive sites within the wall fabric, a change in electrochemical charge density is effected which can be monitored in terms of heavy-metal-binding capacity. Most monoaldehyde fixatives and osmium tetroxide render the wall as reactive, or less reactive, to uranyl acetate as unfixed walls, whereas glutaraldehyde can significantly increase the binding capacity.


Incidence and relationship of group D streptococci with other indicator organisms in meats. Raw and processed meats were analyzed for presumptive group D streptococci using KF streptococcus agar. Counts were compared with coliform, presumptive Escherichia coli, and Enterobacteriaceae counts but no meaningful relationships were observed. Results indicated that group D streptococci and E. coli type I were principally contaminants from the packing plant, rather than at retail level. The predominating group D streptococcus in both beef and pork cuts was Streptococcus faecalis, while in processed meat (bologna), the predominating group D streptococci were Streptococcus faecium var. durans and Streptococcus faecium. Streptococcus bovis was not detected among the isolates from any meat samples. Marked differences were noted in numbers of group D streptococci in processed meat from different manufacturers. The results did not support the use of group D streptococci as alternative indicator organisms for meats. However, the association of group D streptococci with packing plant contamination may prove to be of value.


[Production of vitamin B 12 by a blue-green alga]. In this study, we have been able to demonstrate the production and liberation of vitamin B12 (up to 16.5 pg/ml after 31 days of growth) by a blue green alga. Anabaena flos-aquae, in non-axenic and axenic cultures. This finding has a relevant ecological significance since it shows that, like bacteria, blue green algae may play a role as producers of vitamin B12.


Breast-feeding protects against infection in Indian infants. A retrospective study was undertaken at two isolated Manitoba Indian communities to determine whether the type of infant feeding was related to infection during the first year of life. Of 158 infants 28 were fully breast-fed, 58 initially breast-fed and then changed to bottle-feeding and 72 fully bottle-fed. Fully bottle-fed infants were hospitalized with infectious diseases 10 times more often and spent 10 times more days in hospital during the first year of life than fully breast-fed infants. Diagnoses were mainly lower respiratory tract infection and gastroenteritis. Gastroenteritis occurred in only one breast-fed infant. Breast-feeding was strongly protective against severe infection requiring hospital admission and also against minor infection. The protective effect, which lasted even after breast-feeding was discontinued, was independent of family size, overcrowding in the home, family income and education of the parents. Measures to achieve breast-feeding for virtually all infants, particularly in northern communities, should be given high priority.


Management and results of localized Ewing's sarcoma. Seventy-six patients with localized Ewing's sarcoma who received primary treatment at M.D. Anderson Hospital from 1948 through December 1975 were reviewed. Patients have been divided into four groups according to the different treatment regimens they received: Group I, moderate dose radiotherapy alone; Group II, high dose radiotherapy alone; Group III, radiotherapy plus vincristine and cytoxan; and Group IV, radiotherapy plus vincristine, Adriamycin, cytoxan and actinomycin. The problem of local recurrence appears to be solved with combined chemotherapy and radiation therapy with only one of 36 patients having a recurrence at the primary site in Groups III and IV. Multimodal therapy is the preferred treatment to obtain control of the primary lesion by radiation therapy while preserving good function. However, the major cause of failure remains distant metastases, 19 of 36 (53%) in Groups III and IV. In addition, 4 of 10 patients who have survived over 5 years have developed osteogenic sarcoma.


Solitary plasmacytoma of bone vs. extramedullary plasmacytoma and their relationship to multiple myeloma. Solitary plasmacytoma of bone (SPB) and extramedullary plasmacytoma (EMPC) are unusual solitary tumors of plasma cell origin. The clinical differences between these tumors and multiple myeloma are the subject of continued study. This review examines the radiotherapy experience at M. D. Anderson Hospital. Between 1948 and 1977, 12 patients with SPB and 12 patients with EMPC were treated with radiotherapy. Radiotherapeutic doses were most often (22/24) greater than 4,000 rads, at 200 rads per day. In order to qualify as solitary plasmacytoma a disease-free interval of 3 years was required. In the group of SPB, 5 patients of 9 were alive with no evidence of disease (NED) at 3 years, 4 of 5 at 5 years, and none of 4 at 10 years. In the EMPC group, 8 patients of 11 were NED at 3 and 5 years, and 6 of 9 at 10 years. Six patients with SPB developed multiple myeloma compared with only two with EMPC. The results confirm the better prognosis of EMPC and support the theory that SPB and EMPC are two different entities.