Comparative studies concerning the suitability of European hamsters and Syrian golden hamsters for investigations on smoke exposure. The suitability of the European hamster and the Syrian golden hamster for experiments with tobacco smoke has been tested under various conditions by exposure of the animals to total smoke and the gas vapour phase of two cigarettes. Respiration rate, tidal volume and CO-Hb concentrations were measured before and after the exposures. The results demonstrate that the European hamster tolerates smoke inhalation under all conditions studied better than the Syrian golden hamster. In addition, the haemoglobin of the European hamster showed a markedly lower affinity for carbon monoxide than that of the Syrian golden hamster. In the European hamster, therefore, the danger of CO poisoning after repeated exposures to tobacco is much smaller than in the Syrian golden hamster. The European hamster can be regarded as a more suitable model for investigations with highly concentrated and prolonged smoke inhalations.


Pharmacological properties of 6,7-tetramethylene-5-phenyl-1,2-dihydro-3H-thieno[2,3-e](1,4)-diazepin-2-one (QM-6008, thiadipone), a new psychotropic drug. The pharmacological actions of 6,7-tetramethylene-5-phenyl-1,2,-dihydro-3H-thieno]2,3-e](1,4)-diazepin-2-one (QM-6008, thiadiapone), a new psychotropic drug, have been studied. QM-6008 shares many of the psychosedative effects in rodents of benzodiazepines. The electroencephalographic actions in rats and rabbits of QM-6008 and chlordiazepoxide are also rather similar. This new compounds is endowed with clear conflict attenuating properties in an approach-avoidance schedule and can be considered, in consequence, as a new potentially useful anxiolytic drug. In other operant conditioning procedures in rats--Variable Interval and Discriminated Avoidance--QM-6008 induces an increase in response rate which is not generally shared by chlordiazepoxide. The psychopharmacological data collected in the present work leal to classify this new thienodiazepine derviative within the fram of tranquilizers of the benxodaizapine type. The sedative effects of QM-6008 appear howerer to be less potent and the psychostimulatn effects more markde than those ofchlordiazeposide.


[Quantitative determination of the main metabolites of acetylsalicylic acid. 1. A method for the quantitative determination of salicylic acid and its metabolites. Studies in healthy individuals]. For special purposes a combination of methods is described which allows to determine salicylic acid (including acetylsalicylic acid) and its main metabolites in blood plasma and urine separately and quantitatively. Salicylic acid (SA) and salicyluric acid (SU) are extracted from the acidified fluid with ether and afterwards reextracted in an aqueous phase. By fluorometry at different wavelengths, it is possible to differentiate between SA and SU. The conjugates of SA and SU are hydrolyzed with sulfuric acid and then extracted in the form of SA and SU. The four analyses (SA, SU and their conjugates) are of satisfying accuracy and sensitivity. To test the validity of the method in vivo, the concentrations of SA and its conjugates were determined in the plasma and urines of healthy volunteers. By a simplified, but nevertheless accurate, modification of the described method total salicylate in the urine can be determined.


Pharmacokinetic analysis of ampicillin concentration in neonates: comparison of two pharmacokinetic models and of two numerical methods. Ampicillin blood levels after i.m. injection were analysed according to a one and a two-compartment pharmacokinetic model. The two-compartment model gave no substantially better fit to the data than a one-compartment model. To fit the one-compartment model two computer programs based on different numerical methods, were tested. The BMD program gave satisfactory results in all cases.


Conformation and motion of the choline head group in bilayers of dipalmitoyl-3-sn-phosphatidylcholine. The conformation and motion of the choline head group in lipid bilayers above and below the gel-to-liquid crystal transition point are studied by means of deuterium and phosphorus magnetic resonance. For this purpose dipalmitoyl-3-sn-phosphatidylcholine is selectively deuterated at various positions on the choline and glycerol constituents. The residual deuteron quadrupole couplings and the phosphorus chemical-shift anisotropy of the corresponding lipid-water mixtures yield quantitative information on the segmental motions. The choline methyl group is only slightly hindered in its movement, but the motional freedom becomes increasingly restricted the closer the segment is located to the glycerol backbone. The average value of the OC-CN bond rotation angle changes with temperature. Increasing the temperature rotates the choline methyl group into the vicinity of the phosphorus atom. The choline group as a whole is thus characterized by a flexible, temperature-dependent structure. Its orientation in space is not fixed, either parallel or perpendicular to the bilayer surface. Instead all segments execute angular oscillations with varying degrees of restriction around the normal on the bilayer surface. The gel-to-liquid crystal phase transition at 41 degrees is clearly reflected in the deuterium and phosphorus resonance spectra of the choline moiety, while no change is observed at 34 degrees. The calorimetric pretransition at 34 degrees seems not to be associated with a conformational change in the choline group.


On the mechanism of sugar and amino acid interaction in intestinal transport. The influence of amino acids on D-glucose transport was studied in isolated vesicles of brush border membrane from rat small intestine. It is demonstrated that: (a) Uptake of D-glucose by the membranes is inhibited by simultaneous flow of L- and D-alanine into the vesicles. (b) Addition of L-alanine to membranes pre-equilibrated with D-glucose causes efflux of this sugar. (c) The influence of amino acids on D-glucose is dependent on the presence of Na+. (d) The ionophorous agents monactin and valinomycin are able to prevent the transport interaction of D-glucose and amino acids. Monactin is effective in the presence of Na+ without further addition of other cations, while valinomycin is effective only with added K+, in accordance with the known specificity of these antibiotics. (e) The inhibitory effect increases with L-alanine concentration up to about 50 mM after which it levels off. The experiments provide evident that the Na+-dependent sugar and amino acid fluxes across the brush border membrane are coupled electrically.


Electrical relaxation processes in black lipid membranes in the presence of a cation-selective ionophore. The time course of relaxation of the electric current following steps in the applied potential across lipid bilayer membranes has been measured. The membranes were made cation-selective by the addition of nonactin. To permit the measurement of very short time constants a voltage clamp device was developed in order to reduce the charging period to less than 1 musec, regardless of the magnitude of the series resistances in the external solutions. It was possible by this method to establish the presence of two electric processes, which were found to behave differently with respect to temperature, applied potential and external solution conditions. The rapid process (tau approximately 10 musec) was interpreted in terms of the electric parameters of the polar part of the membrane according to the theory developed by Hägglund and Sandblom (T.I.T.J. Life Sic. 2: 107, 1972). The second process (tau approximately 100 musec) showed a behavior consistent with the model of Stark, Ketterer, Benz and Läuger (Biophys. J. 11:981, 1971), which considers the different rate constants involved in the net transfer of carriermediated ion transport across bilayer membranes.


Pneumothorax complicating pulmonary emphysema. Clinical and roentgenographic findings were compared in patients 40 years of age and over and in those under 40 who were treated for acute unilateral pneumothorax. Dyspnea and anxiety were pominent in the older individuals, although pneumothoraces were usually small. Because physical findings were often unreliable, roentgenograms were required. In the presence of pulmonary emphysema, loss of retractility prevented total collapse of the underlying lung. Increased intrapleural pressure caused over-expansion of the chest wall and the depression of the diaphragm without much mediastinal shifting. Partial collapse of emphysematous lobes demonstrated bullae that were not previously obvious. Respiratory failure developed in five patients over 40 years of age, but four of them recovered after relief of the pneumothorax. Mortality for the group was low and related to associated pulmonary diseases.


Chronic lead absorption. Result of poor ventilation in an indoor pistol range. Three cases of mild lead poisoning were discovered among instructors at an indoor pistol range. These cases were characterized by blood lead levels greater than 100 mug/100 ml, free erythrocyte protoporphyrin levels greater than 450 mug/100 ml of red blood cells, abdominal pain, and, in one instance, by slowing of motor and sensory nerve conduction velocity. Exposure to airborne lead produced during revolver-firing and bullet-molding accounted for the lead absorption. Ventilation in the range was inadequate.


Reye syndrome with associated influenza A and B infection. In early 1974, seventeen children were treated for Reye syndrome. Thirteen of these were studied for laboratory evidence of concomitant viral infection. Influenza B/Hong Kong was isolated from the pharynx in four of nine 1974 cases tested. One child had just recovered from varicella. Adenovirus type 2 and respiratory syncytial virus were isolated from two additional patients. Serologic tests for influenza suggested concomitant or recent influenza B infection in ten of 13 of 1971 cases. During February 1975, six children were treated for this syndrome. Influenza A/Port Chalmers was recovered from three in six 1975 cases, and all six showed significant antibody rises to influenza A. These studies suggest that influenza viruses provide a trigger mechanism for the development of Reye syndrome in susceptible children.


Traumatic left ventricular aneurysm. Two cases with normal coronary angiograms. Two young males had traumatic left ventricular aneurysm and normal coronary angiograms. Trauma was penetrating (bullet) in one and nonpenetrating (fist blow) in the other. Recurrent ventricular tachycardia in one and a systemic embolus in the other required aneurysmectomy. The patients are free of symptoms 42 and 21 months after surgery. The presence of traumatic ventricular aneurysm and normal coronary angiograms suggests that myocardial necrosis following chest trauma is often caused by direct injury to the myocardium rather than by injury to occlusion of the coronary arteries. Therefore, coronary artery bypass surgery may be unnecessary or even hazardous in persons with chest trauma and evidence of early myocardial necrosis.


[Infulence of pyrasanone on the index of articular radioisotope (tc99m) uptake in rheumatoid arthritis]. Objective and instrumental appraisal of the therapeutic effect of a new inflammatory molecule (pyrasanone) via radio-isotope and clinical assessment is reported. A highly significant decrease in articular uptake of radiotechnetium was observed when pyrasanone was administered for a short period to 6 subjects with classic or definite rheumatoid arthritis. Clinical assessment after prolonged aministration in 40 patients with various rheumatic diseases with pathogenetice inflammation showed a marked anc clinically important anti-inflammatory (and hence analgesic) effct; tolerance was good and there were no side-effects.


Surfactant in the lung and tracheal fluid of the fetal lamb and acceleration of its appearance by dexamethasone. We measured tracheal fluid production rate and pulmonary surfactant flux in 12 sets of twin fetal lambs of 102 to 135 days' gestation. In nine of these sets we also measured surfactant flux and the concentration of saturated lecithin in the lungs before and during dexamethasone infusion into one of the twins. The average tracheal fluid flow rate was 3.25 ml/kg/hr (SD 1.6) and, relative to body weight, did not change between 103 and 135 days' gestation or during the infusion of dexamethasone. In untreated fetuses, surfactant was detected between 108 and 130 days and its flux gradually increased but remained less than 150mug/kg/hr at 135 days' gestation; the amount of disaturated lecithin in the lung relative to body weight increased 13-fold from 108 to 134 days. From these data we calculated that the minimal rate of synthesis of disaturated lecithin was increased about 4-fold by dexamethasone from 108 to 120 days; this enhancement fell to about 1.8-fold by 135 days. The rate of secretion of disaturated lecithin, as estimated by surfactant flux in tracheal fluid, was increased by dexamethasone throughout the interval of the study.


Ejaculatory pattern in female rats without androgen treatment. Adult female rats receiving long-term estrogen treatment displayed the species-typical motor pattern of ejaculation during copulation. This hormone treatment produced pituitary hypertrophy and concomitant pressure damage to brain areas dorsal to the pituitary, but did not cause clitoral hypertrophy. The demonstration of the ejaculatory pattern in perinatally untreated female rats indicates that the potential for the expression of the ejaculatory or "orgasmic" pattern is not dependent on exogenous androgen at any stage of development and is more widely represented among female mammals than previously believed.


Remnant function after subtotal thyroidectomy for Graves' disease. Although it has been thought that future thyroid function after subtotal thyroidectomy for Graves' disease was intimately related to the mass of the thyroid remnant, long-term functional results have not been clearly established. These questions were examined in a retrospective study of 86 surgical cases with histologically confirmed Graves' disease. Operation resulted in euthyroidism in 76% of cases. Recurrent thyrotoxicosis appeared in 8%. Hypothyroidism developed in 14 patients (16%), all with estimated remnant weights of less than 8 gm. In each case, hypothyroidism was diagnosed within the first postoperative year. The capability for early diagnosis of hypothyroidism, however, is a significant advantage of surgical treatment. Within the range of estimated remnant weights, a significant and negative correlation was found between remnant size and the subsequent development of hypothyroidism, but neither euthyroidism nor recurrence of thyrotoxicosis could be correlated with remnant size. In the individual patient, therefore, estimated remnant size is of limited value in predicting future thyroid function.


Elevation of serum immunoglobulin M (IgM) level in patients with brain tumors. Sera of 43 brain tumor patients were analyzed along with various other control groups for serum immunoglobulin, cold and sheep cell hemagglutinin and a heterophile type antibody. Serum IgM and heterophile antibody (non-Forssman type) were found elevated in brain tumor and various acute viral meningoencephalitis groups. Serum IgA and IgG levels were essentially unchanged. IgM levels appear to be particularly stimulated by the meningioma class because the removal of tumor from the host was accompanied by its return to normal range. This system of IgM measurement may prove to be a valuable tool in preliminary diagnosis of brain tumors.


Intracerebral toxoplasmosis presenting as a mass lesion. Intracerebral toxoplasmosis is an uncommon condition and for it a produce an intracerebral mass is rare. A case is reported in a patient also suffering from lymphosarcoma, who had received immunosuppressive therapy and who had recently had herpes zoster ophthalmica.


The metabolism of tolamolol in the mouse, rat, guinea-pig, rabbit and dog. 1. [3H, 14C]Tolamolol was well absorbed after oral administration to mice, rats, guinea-pigs, rabbits and dogs. 2. The major route for excretion of radioactivity by mice, rats and guinea-pigs was the faeces; in rabbits the major route was the urine. Dogs excreted similar amounts of radioactivity by both routes. Biliary excretion of radioactivity by the rat and guinea-pig was demonstrated. 3. Tolamolol was extensively metabolized by all five species. The major metabolite in mice, rats, guinea-pigs and rabbits was the product of hydroxylation of the tolyl ring, which was excreted as such as the glucuronide and sulphate conjugates. 4. In the dog the major metabolite was the acid resulting from hydrolysis of the carbamoyl group. This acid was also excreted by the rabbit, but was only a minor metabolite in the other species studied.


Age and individual differences in infants' learning abilities. To assess age and individual differences in infants' learning abilities the method of conditioned head turning toward the source of milk was applied to 20 infants at two different age periods - one month and four months. The results indicate that with increasing age the conditioned reflex is established more speedily in the first six months of life with a corresponding shortening of the mean latency. Extinction, however, occurs more speedily in one-month-old infants than in four-month-old infants. There are no differences in the mean latency of positive reaction during extinction and establishment of the conditioned reflex in both age categories. Considerable individual differences were ascertained in the rate of establishment and extinction of the conditioned reflex. Differences in the rate of extinction of the conditioned reaction appeared to be relatively stable.


An automatic static perimeter, design and pilot study. An instrument for automatic static perimetry at 14 points across a meridian was constructed and used in a pilot study. The machine was controlled by a computer and the subjects responded to the light stimuli by pressing one of two push-buttons. Most people investigated readily understood the rules at testing. "Blunders" made by the subject were, however, so common, that allowance had to be made in the test programme for a number of mistakes. The test stimuli should be exposed in random order. When the zone of uncertain seeing is wide - this occurs especially in untrained subjects - a simple test logic gives a fairly large variation. An improvement in reproducibility is then obtained by applying a more complicated logic, though at the cost of an increased duration of the test session.


Smoking and intraocular pressure.

Smoking and intraocular pressure. The intraocular pressure - as measured by Schiötz tonometer - was studied in 378 smokers, 151 ex-smokers, and 495 non-smokers. It was found the the three groups have practically the same distribution of intraocular pressure, and that the latter has non relationship to the smoking habit.


Permeability of rabbit corneal epithelium to horseradish peroxidase after the influence of benzalkonium chloride. A tight barrier against permeation of horseradish peroxidase into the corneal epithelium exists at the corneal surface adjacent to the tear film. The present light and transmission electron microscopic study reveals that the cationic surfactant, benzalkonium chloride, which is commonly added to eye drops as a preservative, breaks down this barrier. Lysis of the cell membranes was demonstrated, resulting in a leakage of the tracer into and underneath the superficial cells. The grade of cellular destruction caused by benzalkonium chloride was dependent upon the concentration and exposure time of the drug.


The effect of benzalkonium chloride on the electropotential of the rabbit cornea. The effect of benzalkonium chloride on the electropotenial of the cornea has been examined. The anterior surface of the in vivo or in vitro cornea was exposed to various concentrations of the surfactant, from 0.005% to 0.02%, for either 1 or 2 min. The initial effect is a hyperpolarization lasting up to 30 sec, followed by a rapid fall in potential difference with a subsequent recovery. The degree of potential difference decrease and the recovery rate was dependent upon both the concentration of the detergent, and the exposure time. There is excellent correlation between the previous anatomical and physiological studies on tracer penetration across the in vivo and in vitro cornea and our present work. The data indicate that benzalkonium chloride acts by breaking down the physiological and anatomical diffusion barrier to solute and solvent which is located in the outer layer of the epithelium.


Intramuscular rabbit anesthesia ketamine hydrochloride and mebumalnatrium (NFN). A safe and easy combination. For the attainment of deep anesthesia in rabbits, a combination of ketamine hydrochloride (Ketalar) and mebumalnatrium (NFN) (Nembutal), administered intramuscularly, was evaluated. The anesthetic effect was satisfactory and no deaths or side effects were recorded.


Concentrations of some metabolites in the aqueous humour of human senile cataractous eyes. Glucose, pyruvate, l-lactate, and citrate concentrations of the aqueous humour were examined in 51 patients aged 54-87 years with senile cataracts, and in one patient with a clear lens. Furthermore, the ratios of glucose in aqueous/glucose in plasma and citrate in aqueous/citrate in plasma were investigated. Among the parameters recorded, age dependence could only be demonstrated for glucose concentrations in the aqueous humour, as this parameter decreases with increasing age. However, a chance significance because of multipleignificance tests cannot be left out of account in this material. The parameters in question are unfit for an evaluation of possible changes in the energy metabolism of the lens in relation to cataract development because no significant differences were found between concentrations and ratios in different clinical types of cataract. Also, considerable ranges of variation were recorded.


Objective recognition of abnormal isopters. We have examined the hypothesis that normal and only normal isopters of the central visual field are elliptical, except for random deviations. Statistical methods capable of recognizing small deviations from elliptical shape have been developed. Theses methods have been devised to meet the special demands of clinical testing of visual fields and require only simple calculations. The qualities of these methods were examined by theoretical analysis, by large-scale simulations, and by experimental comparison with subjective evaluations. The new methods proved very useful in these examinations. A clinical study using a servo-equipped Goldman perimeter demonstrated that normal central isopters are elliptical enough for the proposed test characteristic to be of practical value in clinical work. The method allows an objective recognition of abnormal isopters with a high degree of sensitivity.


Quadrant sparing of the macula.

Quadrant sparing of the macula. Macular sparing is an indisputable clinical finding in cases of hemianopia. Several explanations have been given, some of which are not mutually exclusive. Eight cases, encountered in a study of more than 100 hemianopias, are presented, in which the sparing has the form of a quadrant, either upper or lower. The explanation in these cases seems to be the double vascular supply to the tip of the occipital pole. It is stressed, however, that macular sparing may have a different explanation in other cases.


The vitrophage in ocular trauma. Report of 15 cases. Fifteen patients underwent a pars plana approach vitrectomy and membranectomy after acute and chronic injury. In all cases the lens fragments, hemorrhage, and secondary membranes were removed restoring clarity to the ocular media. In two cases the final visual acuity was limited by preexisting retinal injury. The advantages of pars plana approach are the small incision and minimal ocular manipulation during surgery. In the presence of retinal injury, diathermy during the procedure or photocoagulation within 24 hours is also indicated.


Anterior chamber depth and primary angle-closure glaucoma. II. A genetic study. The genetics of primary angle-closure glaucome (a.c.g.) was studied: a) through the prevalence in sibs and children of a.c.g. probands, and b) through the family distribution of the closely correlated axial anterior chamber depth (ACD). The material emerged from an epidemiologic study in Greeland Eskimos. a) Compared with the general population, the observed prevalence of a.c.g. was increased in sibs of a.c.g. probands and the estimated, future prevalence was found to be the same in sibs and children. Age influence prevented a proper Mendelian analysis, but no simple monogenic inheritance seems probable. b) The biometric study showed a relatively shallow chamber in sibs, children, nephews, nieces and grandchildren of a.c.g. probands. Regression analyses revealed a corresponding pattern, also in control families of probands with shallow chambers and in general population families. A heritability of 70% was found, indicating that about two thirds of the age and sex independent variation in ACD seems to be genetic.


Carotid artery aneurysm of granulomatous origin. A native of the Western Caroline Islands presented with a granulomatous aneurysm of the right common carotid artery measuring 7 to 8 cm, which was resected and replaced with a reversed segment of saphenous vein. Adequacy of the collateral circulation to the brain was established by occlusion of the common carotid artery with local anesthesia. This was followed by definitive operation with general endotracheal anesthesia and induced hypertension. Although tuberculosis was the most likely etiologic agent, sarcoid could not be ruled out. Granulomatous aneurysms of the common carotid are extremely rare, and if this case was sarcoid in origin, it is the first such case reported. Only one other similar aneurysm could be found in the literature. Of the various methods of reconstruction of the common carotid artery reported, autogenous reversed saphenous vein is recommended strongly.


Human-to-human transmission of Pseudomonas pseudomallei. Melioidosis, the clinical manifestation of infection with Pseudomonas pseudomallei, has occurred infrequently in American citizens; almost all reported cases have been in Vietnam veterans, usually associated with respiratory disease. A Vietnam veteran from Mississippi developed chronic prostatitis, with no other clinical manifestations, during service in Vietnam, and P. pseudomallei was isolated from prostatic secretions 2 years after his return to the United States. The patient had had sexual contact with four women including his wife since his return from Vietnam. Vaginal and cervical cultures and serum samples were obtained from the four women, and serum samples and cultures of semen were obtained from the patient. Vaginal swabs and semen cultures were negative for P. pseudomallei. The patient and his wife had hemagglutination titers (greater than 640) diagnostic of P. pseudominallei infection. This occurrence of venereal transmission is the first report of person-to-person spread of P. pseudomallei infection.


Structure and actions of saikosaponins isolated from Bupleurum falcatum L. I. Anti-inflammatory action of saikosaponins. Anti-inflammatory action of saikosaponins isolated from the root of Bupleurum falcatum L were examined using female albino rats. The anti-exudative action by granuloma pouch method and the antigranulomatous action by cotton pellet method were demonstrated with i.m. and oral administrations of saikosaponins. The oral administration of saikosaponins in 10 times the dosage of i.m. injection showed almost the same effect. Among saikosaponins isolated from Bupleurum falcatum, saikosaponins a and d, not c, were demonstrated to have anti-inflammatory action. The relationship between structure and action of saikosaponins was discussed. No changes in body weight, adrenal weight, plasma-11-OH-corticosteroid level and hematocrit value were observed.


Comparative study of the activity of boric, benzene-boronic and methyl-benzene-boronic acids upon respiration, general metabolism and systemic hemodynamics of the anesthetized dog. The authors compared the action of boric, benzene-boronic and p-methyl-benzene-boronic acids on respiration and general metabolism and on systemic hemodynamics of the anesthetized dog. They further investigated the effects on the arterial blood pressure in the rat and on the isolated rabbit heart. In the dogs, the three acids were shown to have no significant action on the systolic, diastolic and mean arterial blood pressures and a slight stimulating action on the differential arterial blood pressure. On the contrary, in the rat, all the acids were hypotensive. This action was dose dependent. In the entire animal, as well as in the isolated rabbit heart, benzene-boronic and p-methyl-benzene-boronic acids depressed the myocardial contractility. Whereas boric acid did not act on the heart contractility in situ in anesthetized dogs, it slightly stimulated the myocardial contractility of the isolated rabbit heart. The two acids seemed to be directly active on the myocardial cell. In the anesthetized dog, cardiac performances were first depressed then stimulated. Both benzene-boronic and p-methyl-benzene-boronic acids were ventilatory depressors: they reduced ventilatory output and rhythm, they first decreased then increased total O2 consumption and CO2 production.


The relevance of the osmolarity of the instillation fluid to the effectiveness and toxicity of drugs given by the intraduodenal route--solvent drug influence on the intestinal absorption of drugs. The influence  of movements of fluid from the blood into the bowel lumen (solvent drag) on absorption of drugs from the intestine was studied in conscious rats with surgically implanted duodenal tubes. The drugs were given to the rats by intraduodenal instillation either in isotonic saline or in solutions of mannite having twice or three times the osmotic pressure of the blood. The instillation of hypertonic mannite solutions leads to concentration-dependent exsorption of fluid. The drugs tested were atropine and azoniaspiro compound XVII (azoniaspiro[3alpha-benziloyloxy-nortropan-8,1'-pyrrolidine]-chloride), both of which are poorly absorbed, and phenobarbital and nicotine, both of which are well absorbed. The criteria used to assess the rate of entry of the drugs into the blood stream were "efficacy" and "toxicity". In the case of atropine, azoniaspiro compound XVII and nicotine mortality rate and survival time were measured, while in the case of phenobarbital the latent period between instillation of the solution and onset of the hypnotic effect was used. As the tonicity of the solution was raised, so the concentration-dependent "efficacy" and "toxicity" declined. This meant that the latent period after instillation of phenobarbital lengthened, while after instillation of atropine, azoniaspiro compound XVII and nicotine in solutions of increasing tonicity the mortality fell and survival was prolonged. The only exception to this generalization was found when azoniaspiro compound XVII was instilled in triple strength mannite solution. The reasons for this exception were not ascertained. From the results it is concluded that "solvent drag" is an important factor in the absorption of drugs from the intestine. A stream of fluid passing into the lumen interferes with absorption, and this is reflected in the diminished "efficacy" and "toxicity" of the drug. Another factor which may influence the absorption of the drugs tested is the fall in concentration resulting from the inflow of fluid into the bowel lumen. This tends to flatten the concentration gradient between the bowel lumen and the blood and therefore delays absorption.


Kinetic study of the percutaneous absorption of 1,2,4-3H-labelled flupamesone. Two different creams containing 4,4'-methylene bis-[(9 alpha-fluor-11 beta,21-dihydroxy-16alpha,17alpha-isopropylidenedioxy-1,4-pregnadiene-3,20-dione)-3-methoxy-2-naphthoate] (flupamesone) and triamcinolone-acetonide, both labelled with tritium, were prepared in order to study their absorption kinetics. Results were compared in order to find structure-activity relations following Malkinson's hypothesis. The radioactivity was measured in samples of skin taken at various intervals of time after the application of the cream. The results showed a reservoir of corticoids at 400--700 mu depth and that flupamesone remains longer in skin than does triamcinolone-acetonide.


[Studies on the penetrability of skin (author's transl)]. Subcutaneous implantation of small pieces of plastic foam leads to an exudation, the amount of which can be influenced by experimental conditions. The reproducibility of the amount of the exudation is very satisfactory. For 3 examples the possibility could be demonstrated that after cutaneous application of specific substances these can be analysed in the exudate. This could be done in the case of escin and salicylic acid (which is formed from the applied ester by enzymatic break-down during penetration of the epidermis) using thin-layer chromatography while heparin could be demonstrated disc-electrophoretically. All these substances lead to a statistically significant effect on the exudation process. Combining the 3 substances the therapeutic effect is very long lasting.


[The effect of bencyclan on shear induced platelet aggregation (author's transl)]. The platelet aggregation inhibiting effect of N-[3-(1-benzyl-cycloheptyloxy)-propyl]-N,N-dimethyl-amine (bencyclan, Fludilat) was studied by a new method for quantification of platelet aggregation in viscometric flow. In vitro-addition of bencyclan shows significant inhibition in a concentration of 1 mg/100 ml, total inhibition of platelet aggregation is found at a concentration of 5 mg/100 ml. Peroral application of bencyclan shows no significant effect on platelet aggregation. Intravenous application of bencyclan studied in three patients results in inhibition of platelet aggregation, however, no information about the duration of this platelet-inhibiting effect is yielded.


[Quantitative determination of the main metabolites of acetylsalicylic acid/2nd communication: the concentrations of salicylic acid and its metabolites in patients with renal insufficiency (author's transl)]. Quantitative Determination of the Main Metabolites of Acetylsalicylic Acid / 2nd Communication: The concentrations of salicylic acid and its metabolies in patients with renal insufficiency 9 patients suffering from renal insufficiencies of varing degrees and treated regularly by hemodialysis were given 1.5 g Colfarit (microcapsulated acetyl salicylic acid) as a single dose. The concentrations of salicylic acid (SA), salicyluric acid (SU), further salicylic acid conjugates (SAC) and salicyluric acid conjugates (SUC) were determined in the blood plasma. Likewise urea and creatinine were determined. SA concentration decreased continually and, at the end of the trial (72 h after application), had vanished almost completely from the plasma of most patients. SU increased at first and decreased afterwards. With the exception of the dailysis time SAC and SUC increased during the trial. After 3 days the SUC level was more than 50% of total salicylate (SSS) in most patients. SSS (the sum of SA + SU + SAC + SUC) did not change very much before dialysis, but showed a rather high decrease during the first hours of dialysis. tafter dialysis the SSS levels rose again, apparently as a consequence of a redistribution and of the synthesis of conjugates with decreased tissue affinity. It could be shown that SSS in the blood plasma does not parallel SSS in the whole body. The interindividual variation of SA metabolism as well as the variation of the biological blank values was rather high. The results are discussed with regard to salicylate pharmacokinetics in renal insufficiency and to normal salicylate metabolism.


[Blood-liquor distribution of ampicillin in children with meningitis and without (author's transl)]. The distribution of ampicillin between blood and cerebrospinal fluid was compared in patients with serous meningitis to that in patients without inflammation of the meningeal membranes. Ampicillin was applied as a continuous intravenous infusion. Samples of blood and cerebrospinal fluid were taken after 1,2,4 or 12 h for the determination of the ampicillin concentrations. In both groups of patients the ratio of concentration in cerebrospinal fluid to concentration in plasma rose between the first and second hour after administration and reached a steady-state value thereafter. This steady-state ratio was 0.025 in patients without meningitis and 0.061 in patients with meningitis. These results are discussed in the context of a pharmacokinetic model which describes the distribution of drugs between blood and cerebrospinal fluid.


[Antiviral agents/4th communication: the aromatically substituted carbonic acid amide structure in potentically virustatic compounds (author's transl)]. After the conversion of nitrosubstituted aromatic carboxylic acids (3) into the corresponding carboxylic acid chlorides (2), the N-[1-adamantyl]-carboxylic acid amides 4a, 4b, and 4c may be obtained through interaction with 1-amino-adamantane (1). By the same reaction principle,1-adamantyl acetylchloride (6) obtained from 1-adamantyl acetic acid (7) reacts with 2-nitroaniline (5) to give 1-adamantyl-acetic acid-[2-nitroanilide] (8), which may be converted into 2-[1-adamantyl-methyl]-benzimidazole (10) via N-[1-adamantyl-acetyl-]-o-phenylenediamine (9).


[Relations between the physico-chemical properties, the chemical reactivity and the local anesthetic activity. 32. (Part 1): Binding capacity of a structural protein (wool) to local anesthetics]. 1. The general part deals with the molecular structure of nerve membranes and different local-anesthetic substances as well as the various possibilities of local-anesthetic binding to the nerve membrane. Albumin is usually chosen as a model protein in such investigations, a short extract of which is given. 2. The special part includes the chemical composition and properties of wool macroprotein, the model system used in the present paper. We determined its protein-binding values with cinchocain and tetracain homologues and various known commercial products of local anesthetics (oxybuprocain, parethoxyprocain and tetracain). The binding constants of  these substances on wool protein are shown. 3. A statistically confirmed relationship between the alkyl chain length and the strength of protein binding is given. The free energy per CH2-group ranges between 400 and 500 cal/mol and its larger in cases of low pH values. 4. From the present data, a relationship between the binding constant and the partition constant of each substance of the homologous series studied could easily be observed, which further makes it possible to estimate the characteristic protein-binding constant Kp and also the change in pi-values with the change in free reaction energy of the substances. 5. The large differences in the binding values of the different binding mechanisms, which most probably are dependent on the variable binding affinities of the functional groups as well as on steric factors. But as in the homologous series, only substituents like alkyl amino and alkoxy could be changed, the possibility of some other binding process depending on the alkyl chain length of these groups could not be ruled out. Since the alky groups are non-polar, hydrophobic binding plays a role.


Protection of mice against cancer by immunization with membranes but not purified virions from virus infected cancer cells. The life span of C57/Bl mice inoculated with Lewis lung carcinoma cells was prolonged if the mice were pre-immunized with membranes from these cells infected in vitro with influenza virus. Likewise, BALB/c mice were protected against the malignant tumour WEHI-11 by prior immunization with extracts of cultured WEHI-11 cells which had been infected with influenza virus or Semiliki Forest virus (SFV). Partially purified SFV grown in WEHI-11 cells also protected mice from cancer grafts but neither highly purified SFV nor the glycoprotein from the envelope of this virus protected the mice. It is concluded that SFV-induced immunopotentiation against cancer is not due to covalent linkage of tumour specific transplantation antigen (TSTA) to viral envelope protein but more probably is due to the apposition of viral glycoprotein and cellular TSTA in the plasma membrane of the cancer cell.


Lymphoreticular response to a syngeneic rat tumour: gravimetric and histological studies. Gravimetric and histological studies of lymphoreticular tissues during growth of a syngeneic squamous cell carcinoma in Wistar rats show that the regional lymph node anergy reported previously in this system is associated with replacement of paracortical small lymphocytes by large blastoid cells. The regional node continued to gain weight throughout the period of anergy and showed no atrophy and minimal necrosis. Over the same period the spleen increased in both weight and cytotoxicity.


Cell surface antigen expression on chemically induced murine leukaemias. The immunogenicity of murine leukaemias induced by chemical carcinogens or irradiation in C57Bl or (C57Bl times DBA2) F1 hybrid mice has been studied in vivo by transplantation and in vitro by indirect membrane immunofluorescence (IF) using syngeneic immune or allogeneic immune antisera. Two of 5 leukaemias tested for immunogenicity by assessment of the capacity of syngeneic mice specifically immunized with irradiated (3 Krad) cells to reject small challenge inocula (10(3)-10(4) cells) displayed weak neoantigenicity while 3 were non-immunogenic by this criterion. Antibodies directed against cell-surface antigens of the immunizing cells of 7 leukaemias were not detectable by immunofluorescence tests using sera from the respective immunized mice. H-2 histocompatibility antigens readily identified on normal lymphoid cells using reference Balb/c anti-C57Bl (H-2d anti-H-2b) alloantisera could neither be detected on the majority of transplanted leukaemias nor on 9 primary leukaemias in C57Bl mice induced by N-butyl-N-nitrosourea (BNU). Two of the transplanted leukaemias showed greatly diminished capacity for absorption of alloantibody compared with normal spleen cells. Transplantation to H-2 different recipients, in which the leukaemic cells were invariably rejected, generated a strong humoral antibody response, which was demonstrable against normal lymphoid cells. Failure to demonstrate significant antibody binding by indirect immunofluroescence tests with immune sera, or by absorption, is presented as evidency that H-2 antigen expression is substantially modified on BNU induced leukaemia cells. These findings have implications for the detection of tumour neoantigens on chemically induced leukaemias.


Prediction of local recurrence in colorectal carcinoma: an LDH isoenzymatic assay. In a previous study of colorectal carcinoma, the LDH isoenzyme pattern was determined in 420 tissue biopsies from 36 surgical specimens. The LDHIV/LDHII ratio was increased in tumour tissue, but in a number of cases increased ratios were found in the morphologically uninvolved tissue as well. This was especially pronounced in cases with multiple tumours. The isoenzymatic changes were interpreted therefore as a possible indicator of an early process of malignant conversion. In order to test this hypothesis the original material has been reviewed after observation periods ranging from 5 to 7 years. It appears that the mean LDHVI/LDHII isoenzyme ratio of resection edge biopsies is high (0-92) in patients succumbing with local recurrence, differing significantly (P less than 0-01) from the corresponding mean ratio (0-66) in patients clinically cured.


A spatial and temporal analysis of four cancers in African gold miners from Southern Africa. The pattern of cancer in African gold miners over the 8-year period 1964-71, comprising 2,926,461 man-years of employment was studied. Of the 1344 cancers found, primary liver cancer accounted for 52-8%, oesophageal cancer 12-1%, cancer of the respiratory system 5-4% and cancer of the bladder 4-8%. Analysis of the spatial distribution of these four cancers, both on subcontinental and local scale, showed distinct gradients of occurrence between areas of significantly higher and lower incidence than expected. In the case of primary liver cancer in Mozambique and oesophageal cancer in the Transkei, the spatial distribution reflects closely that found in the general resident population of each territory. The crude incidence rate of primary liver cancer in gold miners from Mozambique dropped sharply over the period of the survey.


A retrospective study of the cancer patterns among hospital in-patients in Botswana 1960-72. Records of approximately 310,000 patients admitted to the 10 hospitals in Botswana between 1960 and 1972 have been studied and details of 1445 patients with malignant tumours abstracted. For the 894 tumours for which there was some supporting evidence--at best histological proof and minimally a clinical description of symptoms--proportional frequencies have been calculated for all sites and comparison made with the findings of other surveys. Cancer of the cervix uteri is overwhelmingly the most commonly occurring malignant tumour and the proportional frequency is among the highest observed in Africa south of the Sahara. Skin tumours are unusually common for Southern Africa in both sexes. In males, penile and prostatic tumours have a relatively high frequency whilst the frequencies for liver and lung are lower than in other parts of Southern Africa. Oesophageal cancer in males has a moderate frequency. Other tumours which show a marked variation of frequency within Africa--Kaposi's sarcoma and cancers of the stomach and bladder--are all low in frequency in Botswana. Tumours which are rare throughout Africa but common in Western Europe and North America--cancers of the colon, rectum and corpus uteri--are also rare in Botswana.


Mucous secretion in rat colonic mucosa during carcinogenesis induced by dimethylhydrazine. A morphological and histochemical study. Our previous studies, in specimens of large intestine resected for carcinoma, have shown abnormal patterns of mucous secretion in areas of apparently "normal" mucosa, where goblet cells produce mainly sialomucins as compared with the true normal colonic mucosa in which sulphomucins predominate. In the present work, large bowel cancer was induced in rats by the administration of 1,2-dimethylhydrazine-2HCl (DMH). We attempted to study the sequential histological and secretory abnormalities which developed in the colonic epithelium during carcinogenesis, and to correlate these changes with those described above in the human. The microscopical and histological lesions observed in the colonic mucosa of DMH treated rats confirmed the findings of other authors and resembled the human colorectal cancer. The earliest changes detected were small foci of hyperplasia accompanied from the 6th week onwards by several foci of dysplasia. Carcinoma in situ appeared at the 15th week and finally invasive carcinoma developed from the 19th week onwards. Changes in the type of mucous secretion, with predominance of sialomucins, were observed in the majority of the areas showing mild to moderate dysplasia whilst the surrounding normal epithelium produced suphated material. Mucous depletion was a common feature in areas of severe dysplasia and carcinoma. These findings correlated well with the similar variations in the mucin composition observed in human colonic mucosa in carcinoma and further supported our previous hypothesis that mucin changes characterized by an increase in sialomucins might reflect early malignant transformation. If this hypothesis proved to be correct, the use of a simple method for the identification of mucins in large bowel biopsies would be of great help in detecting early malignancy.


The influence of site of metastasis on tumour growth and response to chemotherapy. Drug screening trials and general treatment of solid tumours in advanced cancer patients have been concerned only with the site of primary origin, regardless of where metastases might have seeded. Since the environment for tumour growth can differ appreciably at various anatomical sites, an investigation was undertaken to determine the effect of metastatic site on response to chemotherapy. Data from 1961 to 1965 of the screening trials of the Eastern Clinical Drug Evaluation Program were utilized. Response and location data extensive enough for analysis represented 6 sites of primary origin and 6 metastatic site groups, totalling 1687 lesions. Analysis of percentage reduction in tumour size after chemotherapy regimens of up to 60 days revealed a significant amount of variation associated with metastatic sites and a non-significant amount associated with sites of primary origin. Advanced primary tumours showed marked variation in responsiveness and some showed a difference in response to different drug groups. Generally, metastases responded better than the advanced primaries from which they were derived, except for those from breast tumours.


Topographical and ultrastructural alterations of smooth muscle cells lining damaged rabbit aorta. The notion that medial smooth muscle cells can be a source of neointima has been a subject of several recent publications. However, the morphological and topographical changes and the degree of thrombogenicity of these cells once they are lining the vessel wall have not been investigated. We were able to inflict a lesion on the rabbit abdominal aorta which resulted in the sloughing of the intima and several layers of the media. As a result, a layer of medial smooth muscle cells was exposed to the circulating blood. Each of these cells retained its characteristic myofilaments but underwent structural as well as topographical changes. The ribosomal elements were greatly increased and mitochondrial cristea obliterated. The cells were closely approximated, leaving no large gaps to permit escape of blood cells. The subjacent region of the lining cells was somewhat oedematous and the internal elastic lamina was absent. On the luminal surface of the lining cells, remnants of connective tissue fibres were not detected and the cell surface was free of platelets, fibrin strands or other formed elements of the blood.


Adrenal glandular lipids and circulating corticosterone in severely diabetic rats. Young, adult, female Sprague-Dawley rats were fasted for 18 h and then given a single s.c. injection of alloxan (10 mg/100 g body weight) which promptly induced a severe state of diabetes. The animals were killed at frequent time intervals during the 7-day study period in order to record the dynamic changes in their capacity for adrenal steroidogenesis and secretion as measured by fluorometric determination of their circulating corticosterone (Cmpd B) levels as well as by thin layer chromatographic identification of cortical lipid moieties used for steroidogenesis. In addition to severe polydypsia, polyuria and polyphagia, these animals manifested super-normal glucose, triglycerides, free fatty acids and cholesterol in their blood, severe hepatic steatosis, adrenal hyperplasia with lipid depletion from the mineralocorticoid producing z. glomerulosa, thymus gland involution and complete degranulation of their insulin producing islet beta cells. Despite an initial high output of Cmpd B and despite progressive cortical hyperplasia, the serum Cmpd B levels became reduced and many of the animals succumbed suddenly, due most likely to inadequate adrenocortical steroidogenesis. Adrenocortical lipids showed a progressive accumulation of free fatty acids, di- and triglycerides, suggesting that some lipid enzymatic defect could be responsible for the lack of conversion of these lipid entities essential for proper steroidogenesis.


Glomerular localization of aggregated protein in mice; effect of strain difference and relationship to systemic macrophage function. Glomerular uptake of aggregated bovine serum albumin has been studied in 3 different mouse strains. Systemic macrophage activity in the same 3 strains has been estimated by measuring the uptake of carbon particles. The amount of aggregated protein taken up by the glomerular mesangium in each strain appears to be inversely related to the systemic macrophage activity. The possible significance of these findings is discussed.


Role of cellular proliferation in the stimulation of MPS phagocytic activity. Administration of agents such as stilboestrol, endotoxin, C. parvum and zymosan, which enhance the phagocytic activity of the mononuclear phagocyte system, also stimulated cellular proliferation in the liver and spleen as assessed by [13H] thymidine incorporation into these organs, and by autoradiography and the number of mitoses in the liver. Sublethal whole body irradiation abolished the proliferative response to stimulation and also reduced or prevented the increase in phagocytic activity. This latter effect depended on the stimulatory agent used, dose of radiation and time of irradiation with respect to stimulation. The results indicate that cell proliferation is important for the increase in phagocytic activity after administration of C. parvum, zymosan and stilboestrol. Endotoxin, however, appears to act primarily by increasing the capacity of existing cells while at the same time causing cell proliferation. The population of cells responding to zymosan by increased phagocytic activity through proliferation was shown to be resident in the liver.


A scanning electron microscopy study of the endothelial folds in normal and anastomosed common carotid arteries in albino rats. The results of this scanning electron microscopy study show that the endothelial folds of the common carotid arteries of rats are composed of rows of 2 or 3 endothelial cells standing alongside one another on the underlying internal elastic lamina. If anastomosed arteries develop a stenosis the endothelial folds, instead of running longitudinally along the vessels, are rotated through 90 degrees to run transversely. The most important factor causing this change in direction of the endothelial folds is fibrosis of the arterial wall following damage to the blood supply of the vessel during dissection and anastomosis.


A simple method for detecting antibodies to rubella. A simple microplate method of enzyme linked immunosorbent assay for Rubella antibody is described. This Micro-ELISA was compared with haemagglutination inhibition in a study of 188 human sera. The total discrepancy rate between the two tests was only 3-7%.


The scanning electron microscopy of the lysis of fibrin by endothelium. The lysis of a fibrin clot by sheets of endothelium one cell thick was examined by scanning electron microscopy. The unlysed fibrin film appeared as a densely matted surface, without interstices. Commencing lysis appeared as parallel ovals in which the dense matting was altered to show many strands of fibrin criss-crossing the lytic "window" with many interstices. The wall of the lytic crater surrounding the sheets of endothelium revealed a network of fibrin strands. There was condensation of the fibrin at the edge of the hole at the external surface. The floor of the crater contained sheets of endothelium which were recognized by the  shape and size of the nuclei. Flocculent masses of precipitate were present on the endothelial cells in the floor of the crater and occasionally within the lysed ovals at the edge. These may be fibrin degradation products.


Interaction of pig lymphocytes with allogenic kidney cells in vitro. Three lines of epithelial-like cells have been developed from the renal cortices of pigs of known histocompatibility type; the cells have been shown to stimulate the development of cytotoxicity in allogenic lymphocytes cultured in contact with them. For this stimulation to occur, it was necessary for the lymphocytes to be of a type that would show blastogenesis in mixed culture with those from the pig used as the source of the renal tissue; serological differences between stimulator and responder cells, as demonstrated by the use of the cytotoxic alloantisera at present available, did not appear to be required.


The use of Chang cells cultured in vitro for the investigation of cellular iron metabolism. Chang cells have been used as a stable model system for the study of cellular iron metabolism. Iron uptake and the stimulation of ferritin synthesis have been studied together with iron incorporation into the ferritin molecule. About 25-30% of the iron taken up by the cells is found in a soluble, non-haem, non-ferritin form which can be chelated by a number of compounds. Ferritin synthesis is inhibited by the presence of desferrioxamine but not by zinc ions.


Production of defective interfering virus in the brains of mice by an avirulent, in contrast with a virulent, strain of Semliki forest virus. An avirulent strain (A7) of Semliki Forest virus formed nearly as much haemagglutinating and complement fixing antigen in the brains of adult mice as a virulent (V13) strain, yet the infectivities of the brain tissues were different by about 100-fold. It appeared therefore that defective virus particles were fomed by A7 but these were not demonstrated by fluorescent antibody studies. In short-term organ cultures of adult mouse brain, A7 derived from mouse brain showed a typical interference pattern in inoculum infectivity response curves. Furthermore, when mixed suspensions of brain-grown V13 and A7 with equal infectivities were inoculated the inoculum infectivity response patterns showed significant depressions of the V13 response at higher inocula. Such interference was not detected if chick cell grown A7 and V13 were substituted for the mouse grown virus. The avirulence of A7 in adult mice and its rapid protective effect against lethal V13 infection could be due to the production of defective interfering virus particles in the brain.


Pathological changes in pregnant mice infected with Coxsackievirus B3 and given dietary casein hydrolysate supplement. Coxsackievirus B3 infection in pregnant mice leads to a severe pancreatic exocrine insufficiency in the mothers and retarded foetal growth. As a consequence of the pancreatic damage, the animals are rendered incapable of digesting sufficient amounts of dietary proteins to allow maternal liver development to proceed as normal for the pregnant rodent. Faecal nitrogen was increased and the maternal livers were small for the weights of the animals and exhibited a lower than normal nitrogen content. Feeding of additional amino acids and simple peptides in the diet in the form of casein hydrolysate either from before or after virus injection appeared to compensate for the inability of these animals to digest dietary protein nitrogen and allowed maternal liver development and foetal growth to proceed at a rate not significantly different from normal. Although these results apply to infections with Coxsackievirus B3 in pregnancy, they may be relevant to other infections which adversely affect foetal growth by their pathological effects on maternal organs necessary for maintaining optimal foetal growth.


Albinism in Nigeria. A clinical and social study. A study of 1000 Nigerian albinos, all of Negro stock, showed various types of albinism with their different modes of transmission--oculocutaneous, ocular and cutaneous. The much higher incidence among the more settled communities in the south, compared with the more nomadic communities in the north, may be related to greater inbreeding tendencies in the south. The sun and society are hostile to the albinos. Under the tropical sunshine, their melanin-deficient skin develops wrinkles, lentigines, actinic keratoses and epitheliomata from which they may die in early adult life or in middle age. Myopia and other ocular defects retard the progress of many albinos in school and they eventually drop out to seek disastrous menial outdoor occupations. Registering albinos early in life, assuring their families that albino defects are confined to the skin and eyes, advising on protective clothing and sun-screening agents, correcting myopia, assisting with indooor occupations, and early treatment of actinic keratoses and skin cancer should help many albinos to attain social acceptance and a ripe old age.


The effect of the activity of psoriasis on the response to autologous ground skin extract. The effects of autologous extracts of involved and macroscopically uninvolved psoriatic skin on epidermal DNA synthesis have been studied. Previous reports of inhibition of DNA synthesis were confirmed but in a number of patients a stimulant, rather than an inhibitory effect, was observed. The stimulant effect was only present when the disease was in an acute, eruptive phase and was more marked in the involved than in the macroscopically uninvolved skin.


The effect of transfer factor on the in vitro migration of tuberculin stimulated white cells from sarcoid and normal subjects. In the mixed leukocyte migration test, white cells from Mantoux positive and Mantoux negative individuals and from patients with sarcoidosis responded similarly to tuberculin, there being no significant difference in the migration index of the three groups. The cells from patients with sarcoidosis, however, showed a depressed response to phytohaemagglutinin. Transfer factor prepared from an individual with a very positive Mantoux reaction did not have any significant effect on the migration index when added to the white cells from sarcoid patients either in the presence or absence of tuberculin.


Study of a new anomaly of the vermilion border of the lip. A new anomaly of the vermilion border of the lip is described in which part of the vermilion border is replaced by normal skin. Thirty-six cases are reported of this new anomaly, seven of which sought medical advice for this condition.


The absorption of arsenic and its relation to carcinoma. A test dose of arsenic was given to four patients with arsenical carcinoma. Blood arsenic levels, urinary and faecal excretion were measured and compared with those of three normal control subjects. Blood levels and urinary excretion were lower in the carcinoma subjects, suggesting increased storage. Abnormally high retention of arsenic, either medicinal or environmental, may be an individual metabolic trait and may be an important factor in carcinogenesis.


Evaluation of coal tar fractions for use in psoriasiform diseases using the mouse tail test. (I) High and low temperature tars and their constituents. 'High' and 'low' temperature tars were evaluated on parakeratotic mouse tail skin, which was used as a model for the psoriatic keratinization process. The skin was examined histologically for signs of induced granular layers in previously scaly areas; epidermal thicknesses were also measured. It appears that the acidic (phenolic) fractions of coal tars induce granular layers and cause epidermal thickening, whereas neutral constituents alone only cause thickening. It is suggested that tar acids should be further investigated for anti-psoriatic activity.


Evaluation of coal tar fractions for use in psoriasiform diseases using the mouse tail test. (II) Tar oil acids. Mouse tail scale epidermis was used as a model to evaluate anti-parakeratotic activity of tar oil acids (phenols) boiling between 175 and 400 degrees C. Lower boiling phenols seemed to cause more irritation, whereas high boiling phenols caused granular layer induction in formerly parakeratotic skin.


Bio-assay of corticosteroids for topical application. Five commercially available corticosteroid creams were compared with respect to their thinning action on mouse tail epidermis. The greatest effects were produced by Synalar and Sintisone; hydrocortisone had the least action. In addition it was shown that the epidermal thickening induced by vitamin A could be significantly reduced by Ledercort cream. The thinning action of the fluorinated steroids on the epidermis is an important property of these agents when used for the treatment of dermatoses having a hyperactive epidermis. The mouse tail test is considered to be a useful screening test for new topical applications of these substances.


Temperature dependence of water content of stratum corneum. The water content of human stratum corneum has been measured gravimetrically in vitro in relation to relative humidity (RH) and temperature. Water content increased with increasing RH. Stratum corneum water content increased 50% when the temperature was raised from 20 to 35 degrees C at RH below 60%. Temperature dependence decreased with increasing RH until there was essentially no temperature dependence at 90% RH. The conclusion is that temperature changes could significantly affect water content in vivo and pliability of skin at RH below 60%.


Tissue nickel levels and nickel dermatitis. I. Nickel in hair. The determination of nickel by instrumental neutron activation is presented. The 58Ni (n, p) 58Co reaction and high-resolution gamma spectroscopy were employed in these measurements. The nickel content of human hair was found to be more dependent upon the sex of the donor than upon other parameters. Mean values for males (1.01 +/- 0.23 ppm) and females (4.21 +/- 0.54 ppm) agreed with values for the nickel content of hair obtained by other methods in most cases. Nickel sensitivity was not reflected in the nickel contents of hair in the female.


Case report: an asthmatic adolescent and his "repressed cry" for his mother. A case of asthma in an adolescent male is presented. Involvement of the patient in individual, group and family psychotherapy is discussed in detail and particular emphasis placed on the use of family sessions as a therapeutic tool. Alexander's concept of the "repressed cry for the lost mother" is suggested as of aetiological importance.


Effect of adrenaline and guanethidine in reducing intraocular pressure in rabbits' eyes. The effect of Adrenaline 0.5 per cent and Guanethidine 2.5 per cent alone and in combination in reducing intraocular pressure in rabbits has been studied. Six rabbits were used and the efficiencies of 5 different dosage protocols was estimated both as intensity and duration of activity. The 5 dosage protocols were Adrenaline alone; Guanethidine alone; Adrenaline and Guanethidine together; Adrenaline followed in 30 min by ?Guanethidine; and Guanethidine followed in 30 min by Adrenaline. The single formulation containing both drugs appeared to be at least as efficient as the others. Eye drops containing 1 per cent Adrenaline and 5 per cent Guanethidine having sufficient stability for normal use have therefore been formulated.


Quantitative lacrimal scintillography. I. Method and physiological application. Quantitative lacrimal scintillography, using 99mTc sulphur colloid, a high resolution gamma camera, and quantification using a digital computer, is a highly effective way of assessing lacrimal physiology, and of establishing normal flow and drainage values against which pathological cases may be compared.


Quantitative lacrimal scintillography. II. Lacrimal pathology. Quantitative lacrimal scintillography is a useful clinical procedure in assessing lacrimal drainage function in patients with epiphora. Examples are presented of patients with canalicular disease, lid abnormalities, 'functional blocks', and dysthroid disease, in which this technique was valuable. Information regarding the dynamics of a functioning and non-functioning dacryocystorhinostomy is elucidated with quantitative scintillography.


Cortical blindness in cranial arteritis. A case is presented of cortical blindness in cranial arteritis. A clinical diagnosis at the time of presentation was based on the integrity of the pupillary light reflex and the absence of visually evoked responses. Pathological examination confirmed bilateral occipital infarction. This report stresses the need for recognizing this presentation of cranial arteritis and initiating immediate treatment.


Lens fringe in homocystinuria.

Lens fringe in homocystinuria. The lens from a patient with homocystinuria was examined by scanning and transmission electron microscopy. A fringe of zonular remnants was found attached to the anterior lens capsule, and was observed to be composed of masses of short filaments in disarray, together with occasional bundles of normal-appearing zonular filaments. Although a pericapsular membrane (zonular lamella) was not observed, the remainder of the lens capsule and epithelium appeared unremarkable. The lens fringe of white zonular remnants may be characteristic, if not pathognomic, for homocystinuria.


Cataract aspiration-irrigation. Modified needle-tubing-handle combination for push-pull machine. A modification of the needle-tubing combination for the push-pull machine used for aspiration-irrigation of congenital and traumatic cataracts, is described. The new needle-tubing-handle combination is advantageous in that the incorporated handle makes the insertion and maneouvring in the anterior chamber easier and also the whole combination can easily be autoclaved as silastic tubing is used.


The participation of methionine and cysteine in the formation of bonds resistant to the action of proteolytic enzymes in heated casein. 1. The influence of temperature, moisture content and the presence of glucose on the level of available methionine and cysteine in casein was studied. 2. Differences between total and available methionine and cysteine contents of heated casein (90 degrees for 24 h) were determined by an in vitro method. The maximum losses in total and available methionine content were 22 and 51% respectively. The losses in total and available cysteine content were 24 and 100% respectively. 3. The results indicated that for heated casein the release of amino acids by proteolytic enzymes was less complete than for native casein. 4. The results of rat growth assays suggested that diets containing oxidized casein are less well utilized by rats than those containing native casein. The decrease in body-weight of rats receiving the diets containing oxidized casein could be counteracted by the addition of methionine and 20 g unoxidized casein/kg diet. 5. There was a lower level of some available amino acids (determined after enzymic hydrolysis using pancreatopeptidase E (EC 3-4-4-7), leucine aminopeptidase (EC 3-4-I-I) and prolidase (EC 3-4-3-7)), including those essential for the rat, in oxidized casein as compared with native casein. 6. Cysteic acid, in oxidized casein, probably makes impossible the utilization of the amino acids in its neighbourhood. 7. From the differences in the available amino acid contents of the native, oxidized and heated casein it was concluded that the oxidation of casein causes the formation of complexes in the polypeptide chain, resistant to enzymic hydrolysis, but to a much lesser extent than does heating.


Estimation of available methionine and cysteine in proteins of food products by in vivo and in vitro methods. 1. The available methionine and cysteine of proteins were determined by chemical methods after preliminary enzymic hydrolysis. 2. The values for the available methionine and cysteine contents of pure proteins (casein and bovine serum albumin) estimated by chemical methods were similar to those for the total content determined by the method of Moore, Spackman & Stein (1958). 3. Reductions of 15 and 11% respectively, when compared with unprocessed samples, were found in the available methionine contents of sweetened and unsweetened, condensed milks; of roller-dried milk and whey powders, and of mackerel sterilized at 126 degrees, the reductions were 22, 14 and 19% respectively. 4. The available cysteine content of sweetened, condensed milk was reduced by about 32%, whereas for mackerel sterilized at 115 and 126 degrees it was reduced by 64 and 75% respectively. 5. The contents of total sulphur amino acids for these food products did not differ from those for the unprocessed samples. 6. Values obtained for available S amino acid contents by rat bioassay confirmed the results of the in vitro estimations.


The role of energy and fluid imbalance in weight changes found during field work in Antarctica. 1. The effect on body-weight of field work in Antarctica when travelling by mechanical transport has been studied. 2. A steady loss of weight (mean 6-2 kg) was found in field, and there was a slow gain on return to less arduous conditions. 3. Changes in skinfold thickness and hip girth suggested that the loss of weight was due to an energy deficit, and the gain was due to an excess of energy. 4. However, although weight was being lost the energy balance was calculated to be positive. This was probably because the expenditure was underestimated as the result of a failure to take into account: (a) the weight and restriction of heavy clothing, (b) the high-protein diet, (c) the difficulty imposed by the terrain, (d) the intensity of the subjects' response to the cold. 5. Daily weight changes were significantly related to climatic conditions. 6. Daily weight changes were significantly related to fluid intakes, but it was not possible to assess with certainty the contribution made by dehydration to the recorded weight loss. 7. Daily fluctuations in weight were greater than those previously found under more standard conditions.


The relationship of the age of onset of obesity to the success of its treatment in the adult. 1. A survey was done of 2333 men and women who claimed experience of slimming. 2. Their loss of weight was determined from their maximum stated weight and their present weight. The loss of weight was calculated as the percentage of the maximum weight and was related to the stated age of onset of obesity. 3. The results showed that those people in the survey who had been fat since childhood had lost just as much weight as those people who had become fat as adults. 4. These results suggest that the treatment of early-onset obesity may not be an unrealistic objective.


Substrate interactions on the intestinal mucosa: a concept for the regulation of intestinal digestion. 1. The hydrolysis of glycyl-L-leucine, glycyl-L-tyrosine, tributyrin, sucrose, maltose, soluble starch and alpha- and beta-glycerophosphates by everted segments of rat intestine was estimated separately or in combination. 2. A comparative study showed significant interaction between different substrates which affected their digestion. 3. Two types of interaction were identified: products of hydrolysis (1) affected the hydrolysis of homologous substances, e.g. methionine and alanine inhibited glycyl-L-leucine hydrolysis, maltose reduced glucoamylase (alpha-1,4-glucan glucohydrolase; EC 3-2-1-3) activity (intracatenary interactions); (2) interfered with the hydrolysis of a different group of substances, e.g. tributyrin inhibited dipeptidase (glycyl-L-leucine hydrolase; EC 3-4-3-2) and alkaline phosphatase (EC 3-1-3-1), glycyl-L-leucine interfered with the activity of the latter enzyme (intercatenary interactions). 4. Mechanisms of interactions were suggested by the results of a comparison of the extent of inhibition or activation of two enzymes (glycyl-L-leucine hydrolase and alkaline phosphatase) in situ in everted intestinal segments or after solubilization with papain or Triton X-100, and different treatments known to affect allosteric sites of these enzymes. 5. Tributyrin and dipeptides were found to act on alkaline phosphatase as allosteric regulators. A discontinuity of the Arrhenius plot suggested the existence of different enzyme conformations which were re-arranged by tributyrin. 6. Substrate interactions in digestion were found in adult rat, cat, rabbit and hen. Substantial differences were found between classes (Aves and Mammalia), orders (rodents, lagomorphs and carnivores) and between age-groups within an animal strain (in this instance, for the rat). 7. These interactions are thought to be involved in the co-ordination of digestion with intestinal absorption and to regulate the time and site of subsequent hydrolysis.


The effects of phytate on intestinal absorption and secretion of zinc, and whole-body retention of Zn, copper, iron and manganese in rats. 1. The inclusion of phytate (10 g/kg) in a purified diet containing zinc (15 mg/kg) fed to young male rats significantly reduced growth rate and food intake, and promoted a cyclic pattern of food intake characteristic of an uncomplicated Zn deficiency. The decreased growth rate could be accounted for by the reduced food consumption. 2. Rats maintained on a Zn-deficient diet (0.5 mg Zn/kg) were found to have a cyclic pattern of food intake and a very slight weight gain. The addition of phytate (10 g/kg) to the Zn-deficient diet promoted a net loss of mean body-weight. 3. Rats maintained on the Zn-supplemented diet containing phytate excreted significantly more Zn in their faeces than either pair-fed or ad lib.-fed control rats. Rats given the Zn-deficient diet supplemented with phytate excreted more Zn in their faeces than Zn-deficient control rats. 4. Dietary phytate significantly reduced the average daily accumulation (mug/d) and whole-body retention (relative to dietary intake) of iron, copper, manganese and Zn, whether or not the diet was supplemented with Zn. 5. The addition of phytate to the lumen fluid of ligated loops of rat duodenum maintained in situ significantly inhibited 65Zn absorption, compared with the control systems without added phytate. 6. Other studies using ligated duodenal and ileal loops indicated that Zn is secreted into the gut lumen and approximately one-third of this is normally reabsorbed. Recycling of endogenous Zn may be a significant process in the over-all body economy of this trace element. 7. The absorption of 65Zn added to the diet was significantly reduced by dietary phytate. Dietary phytate also reduced the biological half-life of body 65Zn from 61 to 211 h post-administration, possibly by inhibiting reabsorption of endogenous 65Zn and thus promoting a more rapid loss from the body.


The rate of heat loss during fasting in the growing pig. 1. The heat loss for individual growing pigs, whose initial body-weight was 25-40 kg, was measured continuously for periods of 14 or 21 d when they were maintained at an environmental temperature of either 20 or 30 degrees. For each temperature the feeding level before and after a period of fasting was held at either 39 or 45g/kg body-weight per d. Four pigs were given each treatment; food was withheld from three animals for a 4 d period in each experiment, and the fourth pig (control) was fed for the whole experimental period. 2. After the withdrawal of food, the heat loss decreased rapidly and minimum values were recorded during the third and fourth days of fasting. Fasting heat loss was higher at 20 degrees than at 30 degrees and was not significantly dependent on the preceeding level of feeding. The mean values obtained at 20 and 30 degrees were 457 and 380 KJ/kg 0.75 per d respectively. 3. The heat loss during the periods of feeding was significantly higher (P less than 0.05) at 30 degrees than at 20 degrees, and  was also higher at the higher level of feeding. 4. The apparent efficiency of utilization of metabolizable energy for maintenance was calculated as 0.80. 5. It was concluded that the critical temperature for a fasting pig weighing 25-40 kg is approximately 25 degrees.


Metabolism of epsilon-(gamma-L-glutamyl)-L-lysine in the rat. 1. Study was made of the metabolism of epsilon-(gamma-L-glutamyl)-L[4,5-3H]lysine (GL) in the rat. 2. The compound was largely absorbed from the intestine and metabolized. Labelled lysine was incorporated into blood proteins. 3. In an in vitro experiment with everted sacs of rat small intestine, GL passed through the intestinal wall unchanged. 4. The results of comparative tests using homogenates of different body tissues indicated that the kidneys were particularly active in hydrolysing GL. Their activity was nine times greater than that of the liver and eighteen times greater than that of the small intestine.


Studies on the absorption and retention of calcium and phosphorus by young and mature Ca-deficient sheep. 1. A combination of a mineral balance and a radioactive technique has been used to study the effects of increased levels of dietary calcium and phosphorus on the Ca and P metabolism of young and mature wether sheep previously made Ca-deficient. 2. Ca absorption was directly related to Ca intake until an absorption maximum was reached. Young animals absorbed dietary Ca more efficiently than mature animals and had a much higher maximum rate of absorption. Apparent P absorption was directly related to P intake and maximum P absorption was also higher for the young animals. 3. Excretion of Ca in the urine and into the intestine was not altered by changes in Ca intake. Excretion of P in the urine, however, varied considerably and it is suggested that this process may be largely responsible for P homoeostasis. 4. Ca retention was directly related to Ca absorption. Maximum retention was considerably higher for the young animals than for the mature animals and occurred for each group of animals at exactly the same dietary intake as maximum Ca absorption. 5. Ca and P were retained in a constant ratio of 1-25:1 and the results suggested that P retention was controlled by the rate of Ca retention. 6. Bone turnover was higher for the young than for the mature animals. Animals mobilized skeletal Ca when receiving the Ca-deficient diet and replaced their Ca losses when the dietary Ca was plentiful by changing the relative rates of bone resorption and bone accretion. Although bone resorption was mainly responsible for Ca homoeostasis, bone accretion appeared to play some part in the young animals. 7. Maximum retention of Ca occurred just as the rate of bone resorption became zero and was equal to the rate of bone accretion. The lower maximum retention of Ca for mature animals was due to a lower rate of bone accretion. 8. Results suggested that when bone resorption ceased, Ca absorption was regulated by homoeostatic mechanisms at a level just sufficient to maintain maximum retention, and that the maximum rates of absorption measured may not have been true maximum rates. 9. The consequences of a relationship between bone accretion and maximum Ca retention are discussed with reference to older, pregnant and lactating animals.


Mechanisms of heat damage in proteins. 5. The nutritional values of heat-damaged and propionylated proteins as sources of lysine, methionine and tryptophan. The preparation of a propionylated protein is described, and the effects of this treatment on amino acid composition and availability are compared with the effects of severe heat treatment (autoclaving) of a protein. 2. Using chemical analyses, changes exceeding 5% for total tyrosine, histidine, methionine and cystine contents were found after propionylation of the protein. Autoclaving of the protein resulted in changes in total serine, lysine, methionine, cystine and tryptophan contents. 3. Microbiological estimates of total amino acid contents were not in close agreement with the chemical estimates for the autoclaved protein. 4. Fluorodinitrobenzene-reactive lysine content was reduced to almost zero by propionylation, and by almost 40% by autoclaving. 5. Both propionylating and autoclaving protein reduced the amount of lysine available to the chick by about half. In contrast, the availabilities of methionine and tryptophan to the chick were unchanged by propionylation, but were reduced to 0.66 and 0.44 respectively, relative to the untreated protein, by autoclaving. 6. Because of the difficulties of obtaining reliable absolute estimates of amino acid availability using chick growth assays, our interpretation of results is mainly based on relative values. The merits of microbiological estimates of amino acid availability are assessed.


Mechanisms of heat damage in proteins. 6. The digestibility of individual amino acids in heated and propionylated proteins. 1. The digestibilities of protein and amino acids have been estimated by two different techniques: the analysis of faeces (conventional method) and the analysis of ileal contents (ileal technique). 2. Freeze-dried muscle protein was found by both techniques to be almost completely digested. After autoclaving, the digestibility for the same protein was estimated by the conventional and ileal techniques to be 0.65 and 0.57 respectively. 3. Unmodified lactalbumin was found by both techniques to have digestibility of about 0.90. Propionylation of the lactalbumin reduced digestibility to 0.82 and 0.79 as indicated by faecal analysis and ileal content analysis respectively. 4. In general, the digestibilities of individual amino acids in any one protein sample were rather uniform, and reflected over-all protein digestibility. For each amino acid, digestibility, as determined by both methods, was lower for the modified protein than for the corresponding control protein: estimates based on ileal content analyses were consistently lower than those obtained by conventional analyses. The ileal technique was considered to be both more convenient and meaningful. 5. From the results obtained by the ileal technique it appears that reduced digestibility is an adequate explanation for the reduction found in nutritional value of the autoclaved protein. In contrast, for the propionylated protein, reduced digestibility of lysine is only a partial explanation of the low availability of this amino acid as estimated by chick growth assay. 6. In our experiments we found that the type of dietary protein used did influence the amino acid composition of the ileal contents. This was most marked with the least-digestible protein. These findings do not support the views of Nasset (1962).


Proton magnetic resonance relaxation studies on the structure of mixed micelles of Triton X-100 and dimyristoylphosphatidylcholine. Proton magnetic resonance and gel chromatographic studies on mixtures of phospholipid and the nonionic surfactant Triton X-200 have shown that at temperatures above the thermotropic phase transition of the phospholipid and below the cloud point of Triton, mixed micelles are present at molar ratios above about 2:1 Triton/phospholipid. Proton T1 and T2 (from line widths) relaxation times are reported for protons in Triton micelles and in mixed micelles of Triton and dimyristoylphosphatidylcholine at a molar ratio of 3:1 Triton/phospholipid. The T1 values and their temperature dependence and the activation energies of the various Triton proton groups appear to reflect internal motions of the Triton molecules in the micelle. Measurements of the T1/T2 ratio and frequency dependence (55-220 MHz) suggest that the hydrophobic tert-butyl group in Triton is observed under extreme narrowing conditions. The T1 and T2 values of Triton are unchanged in the presence of phosphatidylcholine. The T1 values of various protons of dimyristoylphosphatidylcholine in mixed micelles are similar to those reported for the phospholipid in sonicated vesicles, which are used as membrane models, and presumably the same coupled trans-gauche motions dominate. The T2 values for the terminal methyl and choline methyl protons in the phospholipid are longer than those reported for these groups in vesicles. Hence, the motion of the phospholipid in the mixed micelles appears to be less restricted than in vesicles. T1 measurements in H20/D20 mixtures are consistent with the idea that water does not penetrate the hydrophobic core of the mixed micelles, while water does solvate the polar oxyethylene and choline methyl groups. Titration with Mn2+ confirms that the oxyethylene and choline methyl groups are on the exterior of the mixed micelle while the hydrophobic groups are located in the micellar interior.


Ion binding by X-537A. Rates of complexation of Ni2+ and Mn2+ in methanol. The rates of complexation are studied through the effects of the paramagnetic ions upon the magnetic resonances of three of the proton species in X-537A = XH. For the dissociation of the complex MX+ leads to M2+ + X- at 25 degrees the rate is (2.4 plus or minus 0.4) x 10(2) sec-1 for Ni2+ and in the range from 2 x 10(4) to 1 x 10(6) sec-1 for Mn2+. For the Ni2+ complex the activation parameters are also determined and discussed in terms of the details of the process. The difference in rate constants found here is much greater than the difference in the dissociation constants.


Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. We describe the use of polyacrylamide gel electrophoresis to estimate chain lengths of double- and single-stranded DNA molecules in the size range 20-1000 base pairs (or nucleotides). Double-stranded DNA molecules of known length produced either by organic synthesis or by restriction endonuclease digestion of viral DNAs were used as standards. The relative electrophoretic mobilities of these standards were examined on both nondenaturing (aqueous) polyacrylamide gels and on denaturing gels containing 7 M urea or 98% formamide. Electrophoretic mobility of DNA is a linear function of the log of molecular weight if appropriate conditions are used, although exceptions are noted. Chain lengths can be conveniently estimated by using as standards bacteriophage gamma DNA restriction fragments or commercially available tracking dyes.


Isolation of a nuclear ribonucleoprotein network that contains heterogeneous RNA and is bound to the nuclear envelope. Rapidly labeled polydispersed nuclear RNA is part of a ribonucleoprotein (RNP) network which in turn is tightly bound to the nuclear membrane. The membranous attachment, therefore, established a connection between chromatin and cytoplasm. The ultrastructure of the RNP network comprises fibrils and granules similar to those observed in intact nuclei. When bound to the nuclear membrane it has the composition of 63% protein, 14% RNA, 0.4% DNA, and 22.6% lipids. The proportion of lipids diminishes to 2.2% when nuclear membrane is not present. Chromatin, nucleoli, and ribosomes are minor contaminants since histones and ribosomal proteins are not detectable in polyacrylamide gel electrophoresis. Nuclear disruption at high pressure in a French pressure cell causes fragmentation of the RNP network into a series of polydispersed RNP particles. Fragmentation can be prevented by using mild pressure, or by disrupting nuclei with high salt buffer and digesting the dispersed chromatin with deoxyribonuclease. A RNP network, almost free of membrane, is also obtained if the nucleus is deprived of its envelope by treatment with Triton X-100. Since no polydispersed RNP particles are found following dissolution of the nuclear membrane, it is assumed that the particles are components of the RNP network whose fragmentation occurs as a consequence of two processes: (a) activation of nuclear nucleases and (b) shearing forces.


Surface polypeptides of the cultured Chinese hamster ovary cell. The organization of the plasma membrane of logarithmically growing Chinese hamster ovary (CHO) suspension cells has been probed using surface label techniques in conjunction with subcellular fractionation and sodium dodecyl sulfate gel electrophoresis. Five components of apparent molecular weights 137,000, 121,000, 97,000, 67,000, and 57,000 have been shown to be exposed at the outer surface of the cell. These components fully meet the criteria of being (a) reactive with two or more surface label reagents, (b) enriched in a purified plasma membrane fraction, and (c) sensitive to proteolytic digestion of intact cells. Three other components of molecular weights 200,000, 44,000 and 30,000 are also reactive with certain surface label reagents, but fail to meet other criteria for cell surface components. Two polypeptides of molecular weights 180,000 and 37,000 are substantially enriched in the plasma membrane fraction, but are unreactive with surface label reagents. The organization of the CHO cell membrane and the applicability of surface label techniques to cultured cell systems are discussed.


Methylated messenger RNA in mouse kidney. Polyadenylated messenger RNA from mouse kidney labeled in vivo exhibited a pattern of methylation distinct from that of rRNA and tRNA. After mice were given L-[methyl-3H]methionine, 4% of the polyribosomal RNA label was bound to oligo (dT)-cellulose; 20-24% of orotate- or adenine-labeled polyribosomal RNA eluted in the poly(A)+ RNA fraction under similar conditions. [3H]Methyl radioactivity was not incorporated into low molecular weight (5-5.8 S) rRNA, indicating the extent of nonmethylpurine ring labeling was negligible. [3H]Methyl-labeled poly(A)+ RNA sedimented heterogeneously in sodium dodecyl sulfate containing gradients similarly to poly(A)+ mRNA labeled with [3H]orotic acid. Based on an average molecular length of 2970 nucleotides, renal mRNA was estimated to contain 8.6 methyl moieties per molecule. Analysis of alkaline-hydrolyzed RNA sampled by DEAE-Sephadex-urea chromatography provided estimates of the relative amounts of base and ribose methylation. Although 83% of the [3H]methyl radioactivity in rRNA was in the 2'-0-methylnucleotide fraction, no methylated dinucleotides were found in mRNA. In poly(A)+ mRNA 60% of the [3H]methyl label was in the mononucleotide fraction; the remainder eluted between the trinucleotide and tetranucleotide markers and had a net negative charge between -4 and -5. The larger structure, not yet charcterized, could result from two or three consecutive 2'-0-ribose methylations and is estimated to contain 2.6 methyl residues. Alternatively, the oligonucleotide could be a 5'-terminal methylated nucleotide species containing 5'-phosphate(s) in addition to the 3'-phosphate moiety resulting from alkaline hydrolysis. Either structure could have a role in the processing or translation of mRNA in mammalian cells.


Segregation of chlorophyll a incorporated into lipid bilayers. Absorption and fluorescence spectra are reported for chlorophyll a incorporated into a number of aqueous phospholipid dispersions. Absorption spectra show that in dipalmitoylphosphatidylcholine bilayers, monomeric and oligomeric forms of chlorophyll a are present in both the gel and liquid crystalline phases. The formation of aggregates of chlorophyll a is reflected in the fluorescence spectra by a marked concentration quenching. In bilayers conatining small proportions of chlorophyll a, a marked increase in aggregation occurs at the transition temperatures that can be detected calorimetrically. At higher concentrations (greater than 1 chlorophyll:100 lipid), the "pretransition" is abolished in the phosphatidylcholines, and the main transition is broadened, consistent with an orientation for the chlorophyll a with the chlorine ring in the head group region and the phytol chain in the fatty acid chain region of the bilayer. In mixtures of saturated and unsaturated lipids, there is no preferential segregation of the chlorophyll a into the unsaturated lipid.


Regulation of ornithine decarboxylase in 3T3 cells by putrescine and spermidine: indirect evidence for translational control. Addition of putrescine of spermidine prevents the increase in ornithine  decarboxylase activity in cultures of 3T3 cells brought about by pituitary growth factors and results in a rapid, specific, and reversible reduction of enzyme activity in cultures previously stimulated by the growth factors. These effects are not due to polyamine toxicity and do not require other organic medium components. The amines apparently share a single carrier-mediated transport system in 3T3 cells. Methylglyoxal bis(guanylhydrazone), an inhibitor of spermidine synthesis from putrescine was found to also inhibit uptake of each amine. Studies with this drug indicate that each amine is effective without further metabolism. Since ornithine decarboxylase activity decays more rapidly in the presence of each polyamine after addition of camptothecin, the major locus of amine action appears to be in the cytoplasm. However, direct inhibition of the enzyme in vivo by assimilated amines appears to account for at most a small part of the reduction in activity, a conclusion supported by the inability to recover activity in vitro. Also, neither amine seems to act by accelerating enzyme inactivation. When amines are removed from the medium, the subsequent recovery of enzyme activity is totally prevented by trichodermin, an inhibitor of protein synthesis, but is only slightly reduced by camptothecin. It is suggested that both putrescine and spermidine reduce ornithine decarboxylase activity by selectively inhibiting translation.


Isolation and structure of a cross-linked tripeptide from calf bone collagen. A cross-linked tripeptide has been isolated from alkaline hydrolysates of NaB3H4-reduced calf bone collagen. The peptide contains dihydroxylysinonorleucine, the most abundant cross-link in bone collagen, and it has a single N-terminal proline and a single C-terminal valine. These amino acids are in peptide linkage with the cross-link, in a trans configuration with respect to the secondary amine.