Cardiac autonomic blockade in exercising dogs.

Pharmacological blockade of autonomic control of the heart was studied in dogs performing mild, moderate, and severe running exercise on a level treadmill. The dogs were studied without drugs, after atropine, after propranolol, and after both atropine and propranolol. As compared with results without drugs, cardiac denervation resulted in elevated resting heart rate (+45 beats/min) but reduced heart rate during moderate (-17 beats/min) and severe exercise (-47 beats/min); no change in cardiac output at rest or during mild exercise but decreases (-23% and -25%) during moderate and severe exercise; and reduced first derivatives of left ventricular pressure at rest (-24%) and during exercise (-35, -41, and -52% for mild, moderate, and severe loads, respectively). Cardiac denervation did not alter end-diastolic left ventricular diameter but significantly increased end-systolic diameter during exercise. It is concluded that blockade of autonomic control of the heart diminishes cardiac output during exercise by reducing heart rate and myocardial contractile force but does not alter cardiac output at rest.


Effect of breathing helium-oxygen on static lung volumes and lung recoil in normal man.

Static lung volumes and static elastic recoil pressure (Pel) were measured in normal subjects breathing air and 80% helium plus 20% oxygen (He+O2). In 22 subjects, He+O2 produced small but significant increases in total lung capacity (TLC) (mean 0.11 liter, P less than 0.001) and residual volume (mean 0.10 liter, P less than 0.01) without change in vital capacity or functional residual capacity. The mechanisms for this change are obscure. In 10 subjects, breathing He+O2 had no significant effect on Pel (paired t-test) at any lung volume measured (50-80% TLC). In one subject, Pel at 70 and 80% TLC was significantly higher on air than on He+O2 (unpaired t-test, P less than 0.05). Because changes in lung volumes and lung recoil were small, we concluded that these effects do not negate the clinical utility of He+O2 flow-volume curves.


Prostaglandin synthesis inhibition restores hypoxic pulmonary vasoconstriction.

Hypoxic pulmonary vasoconstriction in blood-perfused isolated dog lungs progressively diminishes with repeated hypoxic challenges. We investigated the role of prostaglandins in effecting the decay of the hypoxic response by using a double perfusion preparation that could separately perfuse the right and left lungs of a single dog. Degeneration of this response was reversed by the addition of prostaglandin (PG) synthesis inhibitors, aspirin, or indomethacin. Various PG's known to be produced by the lung (PGE1, PGE2, and PGF2alpha), were infused, and only PGE1 abolished hypoxic pulmonary vasoconstriction. Since other workers have shown that lungs can synthesize and release PG's in response to various stimuli, we postulate that PGE1 synthesis in isolated lungs may increase and thereby cause the degeneration of the hypoxic response. The addition of aspirin or indomethacin could inhibit the synthesis of PGE1 and thereby restore hypoxic pulmonary vasoconstriction.


Thermoregulatory responses during competitive marathon running.

To assess thermoregulatory responses occuring under actual marathon racing conditions, rectal (Tre) and five skin temperatures were measured in two runners approximately every 9 min of a competitive marathon run under cool conditions. Race times and total water losses were: runner 1 = 162.7 min, 3.02 kg; runner 2 = 164.6 min, 2.43 kg. Mean skin temperature was similar throughout the race in the two runners, although they exhibited a marked disparity in temperature at individual skin sites. Tre plateaued after 35--45 min (runner 1 = 40.0--40.1, runner 2 = 38.9--39.2 degrees C). While runner 2 maintained a relatively constant level for the remainder of the race, runner 1 exhibited a secondary increase in Tre. Between 113 and 119 min there was a precipitous rise in Tre from 40.9 to 41.9 degrees C. Partitional calorimetric calculations suggested that a decrease in sweating was responsible for this increment. However, runner 1's ability to maintain his high Tre and running pace for the remaining 44 min of the race and exhibit no signs of heat illness indicated thermoregulation was intact.


Immersion diuresis in dogs.

The mechanism of diuresis during the 1st h of immersion was investigated using anesthetized dogs. Four different experiments were carried out. First, left atrial transmural pressure was measured before, during, and after immersion. The data suggest that, although the left atrium may or may not be stretched depending on the conditions of immersion, the amount of diuresis is independent of the amount of left atrial stretch, and therefore a causal relationship between diuresis and left atrial stretch could not be established. Second, bilateral cervical vagotomy was carried out. Immersion diuresis sometimes occurred despite this vagotomy, suggesting that the left atrial stretch reflex was not participating in those cases. Third, negative-pressure breathing was carried out to simulate the negative transthoracic pressure associated with uncompensated immersion. The average left atrial transmural pressure did not change. A slight hemodilution and a moderate diuresis occurred. There was no correlation between changes in left atrial transmural pressure and changes in urine ouput. Fourth, blood studies were done on splenectomized dogs subjected to immersion. Hemodilution occurred and was most marked in dogs which had had their kidneys removed. The hemodilution is sufficient to explain the early phase of the immersion diuresis. The data suggest that, in anesthetized dogs, hemodilution is the probable initiator of diuresis upon immersion and that, in dogs, left atrial stretch is unrelated to diuresis during immersion or negative-pressure breathing.


Response of immunosympathectomized rats to training.

The influence of chronic exercise was investigated with male rats assigned to normal (N) and sympathectomized (IS) groups. Animals in the latter group were injected daily for 5 days with the antiserum to the nerve growth factor beginning 24 h after birth. Training was introduced when the animals were 35--45 days old and lasted for 12 wk. The exercise program used by both groups was progressive in nature and modified in accordance with the performance capability of the IS animals. When IS and N rats performed a standardized treadmill test, the IS rats had significantly higher rectal temperatures than normal animals. Significant training differences were observed in resting heart rate and in mean blood pressures, but only in the N animals. However, both trained groups exhibited significantly less vasoconstricting ability to conditions of lower body negative pressure than their nontrained controls. Varied doses of epinephrine and norepinephrine were injected into both groups and training per se had no significant influence on the responses recorded. However, both IS group had changes which indicated that a supersensitivity to catecholamines had occurred. Although selective training adaptations can occur without sympathetic nerves, it was concluded that an intact nervous system was essential for maximal training effects to occur.


Effect of osmolality on red blood cell viscosity and transit through the lung.

The mean transit times of labeled red blood cells and albumin were compared in eight isolated rabbit lungs perfused with physiological albumin solutions. The osmolality of these solutions was adjusted by altering the concentration of sodium chloride. The ratios of the mean transit times of injected red blood cells to those of albumin increased as perfusion osmolality increased from hypotonic to isotonic and from isotonic to hypertonic levels. This change occurred despite a decline in pulmonary vascular resistance and red blood cell size as osmolality was increased. Red blood cell viscosity (determined with a cone-plate viscometer) increased with osmolality and it was concluded that this change of viscosity impaired the relative rate of red blood cell transit through the lungs. Passage of red blood cells through rigid homoporous membranes appeared to be related primarily to red cell size rather than vascosity. These observations suggest that both red blood cell viscosity and capillary distensibility play an important role in determining the velocity of red blood cells through the capillaries.


Canine pulmonary response to aerosol histamine: direct versus vagal effects.

Histamine, a potent bronchoconstrictor, has been shown to produce bronchoconstriction both directly and by a vagal reflex. To define the relative roles of direct and reflex effects, we studied the pulmonary response of dogs exposed to increasing doses of aerosol histamine before and after vagal blockade or vagotomy. In addition, the relative contributions of aerodynamically large and small airways to the overall response were determined by the measurement of pulmonary resistance on sulfur hexafluoride-oxygen and helium-oxygen mixtures. Histamine aerosol caused a similar dose-dependent increase in resistance of aerodynamically large and small airways and fall in dynamic compliance. The dose-response relationships were not consistently altered by either vagal blockade or vagotomy. The following variables were found not to alter the experimental results: anesthesia, type of aerosol generator, control of breathing during aerosol exposure, spontaneous breathing vs. controlled ventilation after aerosol exposure, cold block of vagi vs. vagotomy. We conclude that 1) histamine aerosol in dogs causes a local dose-dependent constriction of bronchial smooth muscle, and 2) the vagus nerve played a relatively minor role in the pulmonary response to aerosol histamine in these experiments.


Comparative effects of endoperoxide PGH2 and an analog on the pulmonary vascular bed.

The effects of the prostaglandin endoperoxide, PGH2, and a stable endoperoxide analog were compared on the canine pulmonary vascular bed under conditions of controlled blood flow. The analog produced dose-related increases in lobar arterial and small vein pressure when injected into the perfused lobar artery. The increase in resistance in response to the analog was greater during retrograde perfusion than during forward perfusion. These data suggest that the analog increases resistance to flow by constricting pulmonary veins and upstream vessels. The increase in resistance was similar when the lung was perfused with dextran or blood and the response was not different when the lung was ventilated or collapsed. The response was notattenuated by indomethacin, an inhibitor of prostaglandin synthesis. These data suggest that the response to the analog is not related to changes in bronchomotor tone, platelet aggregation, or enhanced prostaglandin synthesis. Although the analog was a potent pressor substance, PGH2 itself produced only small increases in pulmonary vascular resistance. However, PGH2 had greater contractile activity than the analog on isolated helical segments of canine intrapulmonary vein. The present study suggests that endoperoxides may represent an active form of the prostaglandins in the lung but that their effects would be localized in the tissue in which they are formed because of their extremely short biologic half-life.


Dynamic and steady-state respiratory responses to bicycle exercise.

The transient respiratory responses of 10 normal male volunteers to step changes in work load from 0 to 300, 600, and 800 kpm/min were determined by breath-by-breath analysis for tidal volume, minute ventilation, respiratory frequency, end-tidal oxygen and carbon dioxide tensins, oxygen uptake, carbon dioxide elimination, respiratory exchange ratio, and heart rate. Ten experiments were averaged on a 5-s interval basis. Quantitative measures of the dynamics (delay times, half-times, times to peaks, times to plateaus, and plateau amplitudes) are presented. These parameters generally vary with work load and reflect the speed of response of various components of the system. Rapid ventilatory responses were seen at the initiation and termination of exercise; however, they required up to 32.5 s for full development. Repeated runs on three subjects at 600 kpm/min indicate that the experiments are grossly repeatable. The data, at the initiation of exercise, are consistent with the concept of cardiodynamic hyperpnea while the results are not as clear-cut at the termination of exercise.


Computer-based system for analysis of respiratory responses to exercise.

A computer-based system for the determination of tidal volume, respiratory frequency, minute ventilation, oxygen transfer, carbon dioxide transfer, respiratory exchange ratio, end-tidal oxygen, end-tidal carbon dioxide, and heart rate is presented. These variables are first determined on a breath-by-breath basis from data (expired carbon dioxide and oxygen fractions, airflow, and ECG) prerecorded on an FM magnetic type system. The breath-by-breath data are then averaged for each experimental run in 5-s increments. The 5-s increment data from a group of subjects can then be averaged and the SEM determined at prescribed periods of time. For the study of individual respiratory transient we found the 5-s increment data to be more useful than the breath-by-breath data because it has a lesser degree of fluctuation. The system is especially adapted to careful observation of the responses within the first few seconds of a change in work load. Appropriate computer programs are discussed. The results of several experiments are compared with data from other sources and found to be in good agreement.


Atraumatic measuring of pulmonary capillary blood flow in dogs.

Pulmonary capillary blood flow (Qc) was measured by a nitrous oxide uptake technique using a capacitance-plethysmograph to measure pulmonary volume changes. One hundred and four paired Qc and total cardiac outputs (Qt) were measured in 13 normal mongrel dogs. The mean Qc was 0.101 +/- 0.020 1-min -1-kg-1 while the mean Qt, measured by dye dilution, was 0.117 +/- 0.025 1-min-1-kg-1. This technique minimizes subject trauma and provides a method for repeated pulmonary capillary blood flow determinations in acute or chronic studies.


Prediction of PO2 from SO2 using the standard oxygen hemoglobin equilibrium curve.

We have fitted polynomial equations to three major segments of the standard oxygen hemoglobin equilibrium curve (OHEC) using oxygen saturation (SO2) rather than oxygen tension (PO2) as the independent variable. Hill's transformation contributes substantially to the very small residual errors of PO2, less than 0.21 Torr for all points. This characterization of the OHEC is more precise, and for certain applications more conveniently used than other available equations.


Effect of cobalt on the synthesis of liver microsomal cytochromes.

1. The amount of cytochrome b5 was not changed significantly by a single injection of cobalt (60 mg/kg body weight) or by daily injection of cobalt (30 mg/kg body weight) for 4 days or 8 days. On the other hand, the amount of cytochrome P-450 was depressed strongly by both cobalt treatments. 2. The incorporations of [3H]leucine as well as 5-amino[3H]levulinate into cytochrome b5 in cobalt-treated animals were almost the same as those in the controls 5 h after injections of the radioisotopes, whereas the radioactivity of heme labelled with 5-amino[3H]levulinate in microsomal residues after trypsin digestion, which would consist mainly of cytochrome P-450, was higher in the controls than in cobalt-treated animals after 5 h.


Studies on luciferase from Photobacterium phosphoreum. IX. Further studies on the spectroscopic characteristics of the enzyme-FMN intermediates.

The absorption and fluorometric changes of the reaction mixture of luciferase-FMNH2 complex with O2 were re-examined. Rapid formation (k2(app) = 2.0 s-1 at [O2] = 120 micrometer) of an intermediate with a single absorption maximum at 380 nm within the range of 350-550 nm, and a weak fluorescence at 520 nm (less than or equal to 10% of that of FMN when excited at 380 nm) was observed. The absorption and fluorescence spectra and decay rate of the intermediate were estimated by simulation using an analog computer. The decay rate (0.27 s-1 at 20 degrees C) was in agreement with that of an obligatory intermediate of the luminescent reaction previously determined by measuring aldehyde-initiated luminescence. The process of decay of X1 to FMN involved another intermediate X1' with spectroscopic characteristics rather similar to those of FMN.


Inhibition of the biosynthesis of leucomycin, a macrolide antibiotic, by cerulenin.

Cerulenin, an inhibitor of fatty acid synthesis, specifically inhibits the biosynthesis of leucomycin, a 16-membered macrolide antibiotic, in both growing cells and resting cells of Streptomyces kitasatoensis. In growing cells, the production of leucomycin was inhibited as long as cerulenin remained in the culture. In resting cells, 50 percent inhibition was achieved with a cerulenin concentration of 1.5 microgram/ml. Cells in which leucomycin synthesis was inhibited for 9 h remained capable of leucomycin synthesis upon removal of the inhibitor. Cerulenin specifically inhibits the incorporation of [14C]acetate into leucomycin but does not affect total protein or RNA synthesis. The uptake of [14C]acetate was not inhibited under conditions which completely inhibited the incorporation of acetate into leucomycin. Since cerulenin is known to block the condensation of malonyl-CoA subunits in the formation of fatty acids, it can be concluded that the aglycone of leucomycin is synthesized via the polyketide pathway by condensation steps similar to those involved in fatty acid biosynthesis.


Delayed fluorescence from bacteriochlorophyll in Chromatium vinosum chromatophores: characteristics in the presence of o-phenanthroline.

Delayed fluorescence from bacteriochlorophyll in the chromatophores of Chromatium vinosum, a photosynthetic purple sulfur bacterium, was studied in the presence of o-phenanthroline (o-phen) under intermittent illumination. Re-reduction of the photooxidized reaction center bacteriochlorophyll (P+) in the dark interval was accelerated by o-phen. This effect was attributed to the return of electrons trapped in the primary electron acceptor (A) to P+. In the presence of o-phen, the time course of the decay of delayed fluorescence was not coincident with that of the re-reduction of P+. The delayed fluorescence was somewhat intensified at the early stage (within 30 ms) of relaxation in the dark period. Prolonged illumination (longer than 20 ms) or uncouplers such as carbonylcyanide m-chlorophenylhydrazone (CCCP) or valinomycin plus nigericin decreased the intensity of delayed fluorescence and suppressed the stimulation of delayed fluorescence at the early stage. Delayed fluorescence from reaction center-rich subchromatophore particles decayed with a time course identical to that of the reduction of P+ and was not affected by CCCP, in the presence of o-phen. The intensification at the early stage in the chromatophores can be interpreted in terms of charge separation between pairs of P and A, primary electron donor and acceptor molecules, oriented perpendicular to the intact chromatophore membrane, the effect decreasing in parallel with the recombination of P+ and A-.


Purification and properties of four biologically active components of whale luteinizing hormone.

Four biologically active components of luteinizing hormone (LH), designated as LH I, II, III, and IV, were isolated from whale pituitary glands. These components migrated as single bands with different mobilities in disc electrophoresis, and their isoeletric points were at pH 8.1, 8.3, 8.5, and 8.7, respectively. Their amino acid and carbohydrate compositions showed close similarity. The molecular weight of whale LH was determined to be 31,000 by sedimentation equilibrium, and no difference was detected among the four components. The molecular properties of whale LH were compared with the previously reported data for LH from other animal species.


Determination of molecular species of phospholipids from thermophilic bacterium PS3 by mass chromatography.

The molecular species of phosphatidylethanolamine, phosphatidyglycerol and cardiolipin of thermophilic bacterium PS3 were determined by a new and simple mass chromatographic technique. The fatty acyl groups of these compounds, unlike those of usual phospholipids, were all saturated and were mixtures of even and odd numbered carbon chains; their [M-CnH2n+1-COOCH2]+ and [M-Cn+1H2n+3COO]+ peaks overlapped. Thus, the acetyldiglycerides derived from these phospholipids were subjected to a combination of mass chromatography using chemical ionization with NH3 and treatment of the phospholipids with phospholipase A2 [EC 3.1.1.4]. These three phospholipids had similar patterns of diglycerides. Expressed as R1 : R2, %, they were found to be 14:14,0-1%, 15:14,0-4%, 15:15, 22-23%, 16:15, 16-20%, 17:15, 28-32%, 17:16, 6-9%, 18:15, 7-13%, 17:17, 3-6%, and 18:16, 0-1%. These results should be of significance in connection with the high stability of the biomembranes of this bacterium.


Carbohydrate metabolism in Bacillus brevis ATCC 9999.

Bacillus brevis ATCC 9999, which produces gramicidin S, was found to be unable to ferment various carbohydrates such as glucose, galactose, mannose, lactose, maltose, sucrose, and mannitol. This defect in carbohydrate utilization was attributed to the lack of the transport system(s) for various carbohydrates. This organism can take up only fructose and glycerol and form acids. The presence in this organism of the phosphoenolpyruvate: sugar phosphotransferase system for fructose, but not glycerol, as well as kinases for fructose and glycerol was demonstrated.


Binding of cytoplasmic 5alpha-dihydrotestosterone-receptor complex by nuclei from the ventral prostate, liver, and spleen of rats.

Cytoplasmic 3H-5alpha-dihydrotestosterone-receptor complex of the rat ventral prostate was efficiently taken up by nuclei isolated from the liver, the ventral prostate and the spleen. The amount of the complex bound to liver nuclei was approximately 3 times that to prostate nuclei, and a relatively small amount of the complex was taken up by spleen nuclei under the incubation conditions: 0.16 nM complex (2.0 mg as protein) was used. Binding of the complex to liver nuclei was low-affinity and non-saturable, while in nuclei from the ventral prostate and the spleen, high-affinity and saturable binding was observed. Kd of nuclear binding in the latter two tissues was roughly of the order of 10(-10) M. Castration caused a reduction in the binding of the complex to prostate nuclei and replacement therapy with testosterone propionate completely blocked the effect of castration on the nuclear binding. The affinity of high-affinity binding was not changed significantly after castration. No significant effects of castration and testosterone treatment were observed in the binding of nuclei from the other two tissues examined. The physiological significance of the nuclear binding of hormone-receptor complexes in the action of steroid hormones is discussed.


Use of fluorescein mercuric acetate as a probe in studies of thiol-containing proteins.

The reaction of fluorescein mercuric acetate (FMA) with thiol groups is inhibited by barbital (Veronal). However, barbital has no effect on the reaction of 5,5'-dithiobis(2-nitrobenzoic acid) with thiol groups. Complex formation between FMA and barbital was shown by an increase in the fluorescence polarization of FMA in the presence of barbital. These findings indicate that the inhibitory effect of barbital is due to its interaction with FMA. The extent of this inhibition, which depends on the nature of the thiol-containing substance, presumably reflects the structure surrounding the thiol groups. Furthermore, the absorption and fluorescence properties of FMA conjugated with thiol groups are sensitive to the environment. Because of these properties, FMA can be used to investigate the environment of thiol groups.


Photoaffinity labeling of uncoupler binding sites on mitochondrial membrane.

3H 2-azido-4-nitrophenol, a photoactive uncoupler, has been synthesized, and its uncoupling action on oxidative phosphorylation and its binding to the mitochondrial membrane have been studied. The uncoupler bound covalently to the mitochondrial membrane on photoirradiation was 3-4 times that bound reversibly in the absence of light. When irradiation was carried out in the presence of serum albumin, covalent binding was significantly depressed. The pattern of loss of ATP-Pi exchange activity with increasing amounts of the uncoupler suggests that serum albumin prevents the binding of the uncoupler to the functional sites as well. Polyacrylamide gel electrophoresis of photoaffinity labeled submitochondrial particles in the presence of sodium dodecyl sulfate revealed that a 9000 dalton peptide bound high levels of uncoupler. Other proteins in the molecular weight range of 20,000-40,000 and 55,000 were also labeled. Photolysis in the presence of serum albumin or ATP decreased the covalent binding of the uncoupler to all the proteins, but particularly to the 20,000 dalton component. Soluble ATPase and the mitochondrial proteolipid purified from labeled mitochondria showed the presence of label.


Effect of Mg2+ and spermine on the kinetics of Ca2+ transport in rat-liver mitochondria.

Plots relating the initial rate of mitochondrial Ca2+ transport to the Ca2+ concentration (kinetic plots) have a hyperbolic shape in a Ca2+ concentration range of 2.5-100 muM as measured in sucrose or KCl media. In the presence of Mg2+ or a polyamine spermine, which both are competitive inhibitors of Ca2+ binding to low affinity sites at the membrane surface, the shape of the plots becomes sigmoidal. At higher concentrations of these agents linear kinetic plots are obtained as measured in a sucrose medium. In a KCl medium the sigmoidality of the kinetic plots is enhanced by an increase in the Mg2+ or spermine concentration. It is suggested that Mg2+ and spermine affect the kinetics of Ca2+ transport by interfering with Ca2+ binding to low affinity sites of the membrane surface and that the binding of Ca2+ to these sites is the first step of the mitochondrial Ca2+ transport.


Water permeability of elastomers.

In a previous study it has been shown that the free moisture content in freeze-dried BCG vaccine dispensed in vials sealed with rubber stoppers increased during storage. The search for the source of this increase led us to explore the possibility that this additional moisture could originate from the rubber stoppers themselves. Therefore, the water permeability of various rubber stoppers has been studied, and the water content of grey butyl stoppers during some operations (autoclaving, oven-drying, freeze-drying, storage) used in the manufacturing of BCG vaccine has been determined. Our experiments showed: rapid water uptake during steam-autoclaving and rapid water release during subsequent oven-drying of the stoppers; a slow water uptake of the stoppers during freeze-drying and a slow water permeation through the stoppers when vials containing Indicating Drierite were stored in a water-saturated atmosphere. Among 12 types of rubber stoppers tested, the grey butyl stoppers and the silicone stoppers showed the lowest water uptake. Moisture-resistant wrappings decreased significantly the moisture uptake of Drierite. To delay moisture from reaching the vaccine it is recommended that the stoppers employed be as dry as possible.


Acceptability of BCG vaccination.

The acceptability of BCG vaccination varies a great deal according to the country and to the period when the vaccine is given. The incidence of complications has not always a direct influence on this acceptability, which depends, for a very large part, on the risk of tuberculosis in a given country at a given time.


Phygocytosis by rabbit polymorphonuclear leukocytes: the effect of albumin and polyamino acids on latex uptake.

Albumin in low concentrations (0.0001-0.01 weight percent) was found to be an effective inhibitor of phagocytosis of polystyrene latex beads by rabbit polymorphonuclear leukocytes. Polyglutamic acid proved to be an inhibitor of latex uptake at even lower concentrations. Polylysine stimulates phagocytosis, maximal stimulation occurring at 0.002% polylysine. These findings are discussed with reference to the surface properties of latex particles and leukocytes, and particularly with reference to electrostatic interactions in phagocytosis.


Membrane transport by macrophages in suspension and adherent to glass.

We have compared membrane transport of lysine and adenosine by rabbit lung macrophages in suspension and adherent to glass over slips. The rapid sampling techniques employed permitted measurements to be made over intervals as short as 10 seconds. Suspended cells transported both nutrients at a faster rate, although the Km values for the two transport systems remained unchanged. Colchicine, cytochalasin B and adenosine did not interfere with the enhancement.


Pyruvate carboxylase and phosphoenopyruvate carboxykinase in cultured human fibroblasts.

The activities of pyruvate carboxylase and phosphoenolpyruvate carboxykinase were measured in cultural human fibroblasts. Considerable variation was observed in strains derived from different individuals. The activities of both enzymes throughout the culture cycle were measured in two strains. In these strains the specific activities of the enzymes increased during log phase and remained constant during the stationary phase. However, one cell strain exhibited a high activity of pyruvate carboxylase which remained unchanged throughout the culture cycle, suggesting that this enzyme may be regulated differently in different strains of cultured human cells. Familial studies suggest that the observed variations in pyruvate carboxylase activity may be due to a genetic polymorphism.


Protein metabolism during growth of Vero Cells.

Protein synthesis and degradation were studied throughout a growth cycle of Vero cells. The rate of protein synthesis, measured as the rate of amino acid incorporation, reached a maximum at the mid-exponential phase and declined to 10-30% of the maximum in the stationary phase. The rate of protein degradation, measured as the release of radioactive amino acids from uniformly labelled cellular proteins, did not vary in the growth cycle. The amount of protein per cell, measured by an isotopic method, remained constant when normalized to account for the variation in the proportion of actively dividing cells in the cell population during the growth cycle. Cellular protein was determined using this method since it was found that the chemical determination of the amount of protein in the monolayer was not accurate during the early stage of the growth cycle. This was due to a significant amount of serum protein adsorbed to the cells. In this study we were able to show that, in Vero cells, protein synthetic activity is correlated with the rate of cell division, and variations in the rate of synthesis alone are sufficient to meet the changing requirements for cellular protein in a growth cycle.


Applications of gas-liquid chromatography in protein chemistry. II. Determination of amide residues in nanomolar amounts of proteins.

A method for the quantitative determination of amide residues in nanomolar amounts of proteins is described, based on dilute acid hydrolysis at 100 degrees, followed by isothermal gas-liquid chromatography of the ammonia released by on-column neutralisation of the hydrolysate and quantitation by means of a conductometric detector. Amide contents are given for twenty well characterised proteins, as well as for asparagine and glutamine.


Selective concentration of amines from aqueous solutions by a gas purging technique.

Amines are removed from heated, basic solutions saturated with salt by inert gas stripping and selectively trapped as coordination complexes on columns containing copper(II) salts coated on an inert support. An estimate of the amine concentrations at ppm levels can be made by measuring the length of the trapping column colored by the complexes. Concentrations of amines in ppb can be determined by eluting this column with potassium hydroxide and determining the amines in the effluent directly by gas chromatography. The procedure allows amines to be concentrated by a factor of 200 and removes potentially interfering substances from the sample.


Detection of sympathomimetic central nervous stimulants with special reference to doping. II. Comparative study of two adsorption chromatography methods using different XAD resins.

Recoveries of a series of sympathomimetic central nervous stimulants in human urine are measured using either adsorption chromatography on self-filled columns (method A) or with a special resin method suitable for racehorse urine (method B). The Amberlite resins used are XAD-2, XAD-4, XAD-7 and XAD-8 and elution is performed using chloroform. The reported comparative drug extractabilities indicate that in most instances the recoveries follow the sequence XAD-4 greater than XAD-2 approximately XAD-8 greater than XAD-7 using method A. Based on the recovery and purity of the extracts obtained, XAD-8 is preferred for gas chromatographic analysis while XAD-4 is very suitable for thin-layer chromatographic screening work. Comparing the two methods, equally good or better results were obtained with method A for all of the resins studied except XAD-7. Finally, it was found that the effect of refrigerated storage of the resins on the drug extractabilities for central nervous stimulants could be neglected.


Determination of the anti-inflammatory agent carprofen, (D,L)-6-chloro-alpha-methylcarbazole-2-acetic acid, in blood by high-pressure liquid chromatography.

A rapid, sensitive, and specific high-pressure liquid chromatographic (HPLC) assay was developed for the determination of (D,L)-6-chloro-alpha-methylcarbazole-2-acetic acid (carprofen) in blood. The assay involves extraction into diethyl ether from blood buffered to pH 6. The overall recovery of carprofen from blood is 97.3 +/- 5.3% (S.D.), and the sensitivity limit of detection is 100-200 ng/ml of blood using a fluorescence detector with excitation at 240 nm and emission at wavelengths greater than 350 nm. The HPLC assay is amenable to rapid routine analysis of clinical specimens, and the data obtained using this assay showed an excellent correlation coefficient (0.99) compared with a previously published spectrofluorometric assay. The method was used to monitor the blood level-time fall-off profiles in four subjects following single and multiple dose administration of carprofen.


Gas-liquid chromatographic determination of 1,3-dihydro-3-phenylspiro(isobenzofuran-1,4-piperidine), HP 505, in biological fluids using a nitrogen-specific detector.

A gas chromatographic method for the determination of 1,3-dihydro-3-phenylspiro[isobenzo-1,4-piperidine], HP 505, in plasma red blood cells and urine has been developed. HP 505 and internal standard are extracted from basified fluid with hexane and then back extracted into acetic acid. After re-extraction into hexane, HP 505 and internal standard are analysed by gas-liquid chromatography as the N-propionyl derivatives using a nitrogen-specific detector. Concentrations of HP 505 can be measured over the range 2-100 ng/ml plasma. The method has been applied to the analysis of biological fluids from volunteers receiving oral doses of HP 505.


Application of the extractive alkylation technique to the pentafluorobenzylation of morphine (a heroin metabolite) and surrogates, with special reference to the quantitative determination of plasma morphine levels using mass fragmentography.

The pentafluorobenzylation of morphine and related phenolic alkaloids by extractive alkylation is described. The alkylation is performed using tetrabutylammonium as counter ion and ethyl acetate as solvent. Optimum reaction conditions are presented together with the gas chromatographic properties of the derivatives formed. The technique is applied to the quantitation of plasma morphine levels. Using morphine-d3 as internal standard mass fragmentographic analysis of morphine as its pentafluorobenzyl- and pentafluorobenzyl, mono-trifluoroacetyl derivatives is demonstrated, and a case report is presented. Quantitation to a plasma morphine level of 5 ng/ml is readily attainable.


Air sampling methods for s-tetrachloroethane and other related chlorinated hydrocarbons.

An acid-treated Pittsburgh Activated Carbon (PAC) tube for sampling s-tetrachloroethane in air is described that minimizes sample losses through dehydrogenation and dehydrohalogenation reactions. Commercially prepared NIOSH characoal, untreated PAC, and acid-treated PAC tubes are compared as to their effectiveness in stabilizing s-tetrachloroethane. Common homologues and analogues of s-tetrachloroethane cause no interference with the gas chromatographic method. -n addition the effectiveness of the untreated PAC and acid-treated PAC tubes was also evaluated for four different chlorinated aliphatic hydrocarbons.


Simple chemiluminescent detector for the screening of foodstuffs for the presence of volatile nitrosamines.

The construction and subsequent evaluation of an apparatus for the detection of trace amounts of nitrosamines is described. The apparatus consists of a gas chromatograph, a catalytic chamber to generate nitric oxide from eluted nitrosamines, and a chemiluminescent detector to measure the infra-red emission resulting from interaction of this gas with ozone. Examples of the use of the system for determining the nitrosamine concentration in food extracts and other materials are given.


High-pressure liquid chromatographic analysis of aniline and its metabolites.

A high-pressure liquid chromatographic method has been developed for the determination of nanomole quantities of aniline; its metabolites o- and p-aminophenol, phenylhydroxylamine, nitrosobenzene and nitrobenzene; and azobenzene and azoxybenzene which form non-enzymatically by condensation of reactive metabolites. These compounds were separated by reverse-phase chromatography (mu-Bondapak C18 column) and detected spectrophotometrically. The first four components were eluted using methanol-water (15:85) containing 0.26 M ammonium acetate and 0.015 M nickel acetate as mobile phase. The remaining compounds were eluted with methanol-water (50:50). The stabilities of the metabolites were studied electrochemically and results were used in the development of the chromatographic system.


Quantitative determination of naltrexone and naltrexone prodrugs by electron-capture gas-liquid chromatography.

An electron-capture gas chromatographic method has been applied to the determination of eleven ester and ether derivatives of naltrexone believed to act as prodrugs. Standard analytical curves are presented for all prodrugs and quantitation is shown to be possible from 10 ng to 1.5 microgram of each compound. Ester derivatives of naltrexone are hydrolyzed to naltrexone prior to analysis as the perfluoroalkyl esters. Analysis of synthetic mixtures of these with naltrexone demonstrated that quantitation by difference measurements is possible with naltrexone-derivative ratios from 4:1 to 1:10. Ether derivatives are analyzed without hydrolysis. This method is applicable to biological fluids as well as aqueous solutions.


Gas chromatographic determination of pemoline as 5-phenyl-2,4-oxazolidinedione in human urine.

A simple gas chromatographic assay of the psychostimulant pemoline in human urine has been developed. Instead of extraction of the drug from urine, it is hydrolysed to 5-phenyl-2,4-oxazolidinedione with 1 N hydrochloric acid. After the extraction, this compound is methylated with diazomethane and determined by gas-liquid chromatography using a nitrogen-selective detector and a solid injection system. The method has been applied in preliminary human pharmacokinetic studies, by measuring the urinary excretion rate of pemoline following oral administration. At present, the screening procedures for doping control do not involve the detection of pemoline, but the method described can easily be incorporated in such procedures.


Gas chromatographic determination of pemoline in biological fluids using electron capture detection.

A simple and sensitive gas chromatographic (GC) method for the determination of pemoline in biological fluids, utilizing electron capture detection is described. Plasma samples with a pemoline analog added as internal standard were deproteinized with sulfosalicylic acid, and the supernatants were heated at 80 degrees. The pemoline-dione formed was extracted with benzene, and the extract was analyzed on a gas chromatograph equipped with a tritium foil electron capture detector. Poly A-103 (3%) on Gas-Chrom Q (100-120 mesh) packed in a 3-ft. silanized glass column was used as the stationary phase, with nitrogen serving as carrier gass. Under the same GC conditions, benzene extracts of pemoline-dione from acid-hydrolyzed urine samples were analyzed. Us-ng 1 ml of plasma or urine, the lower limit for the assay was about 0.1 microgram/ml. The method is accurate and reproducible, with a relative standard deviation with +/-4%. Mandelic acid (a metabolite of pemoline) does not interfere with the assay.


High-performance liquid chromatographic assay of rat-brain dopamine and norepinephrine.

A high-performance liquid chromatographic method for the assay of dopamine (DM) and norepinephrine (NE) in rat brain has been developed. 3,4-Dihydroxybenzylamine was added to the sample as an internal standard. The isolated amines and the internal standard were subjected to reaction with fluorescamine, separated liquid chromatographically on a glycol-type gel and determined fluorimetrically. The standard deviation of the method was +/-2.7% for DM and +/-5.3% for NE. The contents of DM and NE in rat brain, with or without administration of a tyrosine hydroxylase inhibitor, as measured by the proposed method were always lower than those obtained by the trihydroxyindole method.


Determination of organochlorine insecticide residues in fatty foodstuffs using a clean-up technique based on a single column of activated alumina.

A method is described for the extraction and clean-up of fatty foods prior to the determination of organochlorine insecticides and related compounds. Clean-up is based on the use of a single 22-g column of activity-4 alumina and the method is capable of routinely determining cyclodienes, BHC isomers and HCB at the 5-10 microgram/kg level and DDT-type compounds at 20-30 microgram/kg level.


Determination of hydralazine in tablets by gas chromatography.

A description is given of a gas chromatographic method for the determination of hydralizine in various tablet formulations based on the quantitative reaction of hydralazine with 2,4-pentanedione to yield 1-(3,5-dimethylpyrazole)phthalazine. The stable hydralazine derivative formed is extracted from aqueous solution and chromatographed resulting in a precise and specific determination of hydralazine which correlates well with the U.S.P. XIX titrimetric procedure.


Determination of 2,4-diamino-5-(3,4-dichlorophenyl)-6-methylpyrimidine (BW 197U) in human plasma.

A gas-liquid chromatographic method for the measurement of 2,4-diamino-5-(3,4-dichlorophenyl)-6-methylpyrimidine (BW 197U) in human plasma has been developed. After extraction from alkaline medium into ethyl acetate the compound is injected into a gas-liquid chromatograph and measured using a 63Ni constant-current electron-capture detector. The range of concentrations measured was from 10 ng/ml to 10 microgram/ml in plasma. An internal standard was employed and reproducibility between replicates was found to be good. The method has been semi-automated by the use of an auto-sampler controlled by a minicomputer which also processes the data.


Simple and rapid gas-liquid-solid chromatographic analysis of trace concentrations of acetaldehyde in urban air.

A simple and rapid gas-liquid-solid chromatographic method for the analysis of trace concentrations of acetaldehyde in urban air (in the Nagoya area) was developed, with the use of cold trapping with liquid oxygen. In the analytical main column the conditions were: stationary phase, Triton X-100 (0.4%); support, Carbopack B (60-80 mesh); glass column, 1.5 m X 3 mm I.D.; column temperature, 75 degrees; carrier gas (nitrogen) flow-rate, 50-80 ml/min. In the cold trapping pre-column the conditions were: stationary phase, Tris (2-cyanoethoxy) propane (25%); support, Shimalite (AW, DMCS) (60-80 mesh); glass column, 31 cm X 4 mm I.D.; operating temperature for the trapping, -183 degrees (liquid oxygen temperature); operating temperature for injection of the condensed sample into the gas chromatograph, increased from -183 degrees to +100 degrees for 2 min. The acetaldehyde peak was identified by the disappearance method with a 2,4-dinitrophenylhydrazine-orthophosphoric acid-glass beads column. The ranges and average concentrations of acetaldehyde detected in 13 urban air samples were 1.5-9.6 and 4.7 ppb, respectively.


Extraction procedures for use prior to HPLC nucleotide analysis using microparticle chemically bonded packings.

In preparing nucleotide extracts for HPLC analysis using microparticle, chemically bonded packings, tri-octyl-amine-freon solutions can be used for the extraction of the acid from the solution if the concentration of the amine is carefully controlled.


Acute hormonal response to cigarette smoking.

Preliminary smoking studies using non-smokers were unsatisfactory because the subjects became obviously "ill" with nausea, pallor, sweating, etc. The rise in adrenocorticotropic hormone and growth hormone in these individuals could not be attributed to a cigarette effect as opposed to a nonspecific stress. A small number of female smokers seemed less consistent in their responses than did male subjects. The final study employing male, habitual smokers showed the following results: (1) A sharp rise in circulating cortisol occurred after two cigarettes which was maintained through the 2nd hour and fell slowly after the smoking period. (2) Circulating growth hormones also began to rise after two cigarettes, peaked at 1 hour, and then fell back to control levels while smoking continued. (3) Urinary catecholamines tended to be higher on smoking days than on nonsmoking days, but results were variable and of questionable significance. (4) Luteinizing hormone, follicle stimulating hormone, testosterone, and thyroid-stimulating hormone did not show any significant variations with smoking as compared to non-smoking.


Urinary creatinine as an index of urinary excretion of estrogen in cows prepartum and postpartum.

Urine was collected from 55 cows via indwelling urinary catheters for periods of 12 h on different days (28 days prepartum to 60 days postpartum). Excretion of urinary creatinine (mg/h per kg bodyweight) among Holsteins increased from .94 on day 28 prepartum to  1.14 on day .5 postpartum and then decreased to .82 on days 30 to 45 of lactation. Excretion of creatinine among 12-h collections of urine on different days did not differ for groups of cows within periods prepartum and postpartum, and coefficients of variation within cows were 12 to 13%. Ratio of urinary estradiol (-17 alpha) to urinary creatinine was correlated (.93) more highly with its excretion based on volume of urine excreted than was its urinary concentration (.79) within periods prepartum and postpartum. Ratios of urinary metabolites to urinary creatinine rather than their concentrations should be used to express rates of excretion in cows' urine when urine excreted per hour is unknown. Moreover, daily rates of excretion of estradiol in urine can be estimated [ng estradiol/day = A X ng/mg urinary creatinine x kg bodyweight x 24 h, where A is average excretion of urinary creatinine (mg/h per kg bodyweight)] for respective days prepartum and postpartum.


Rumen bacterial urease requirement for nickel.

Lambs were fed a basal purified diet low in nickel (60 ppb) or the basal diet supplemented with 5 ppm of nickel to determine if rumen bacterial urease was a nickel-requiring enzyme. Two collection periods with lambs fed a diet in which all the nitrogen was supplied as preformed protein (casein) indicated that ruminal urease activity was much lower in lambs fed the low nickel diet. When 1% urea was added to the basal diet, urease activity increased slightly with both treatments; however, bacterial urease activity was still much higher in the lambs receiving 5 ppm of nickel. Ruminal volatile fatty acids were not influenced by dietary nickel. Ruminal urease requires nickel for maximal activity.


A Robertsonian translocation and its effect upon fertility in Brown Swiss cattle.

Metaphase chromosomes derived from leukocyte cultures of peripheral blood were observed in 299 head of Brown Swiss cattle. A Robertsonian translocation (centric fusion) was found in 31 of the animals. Estimated from this small sample 2.4% of animals of the Brown Swiss breed carry the translocation. Chromosome measurements and analysis of G-banded chromosomes proved inconclusive in identification of the homologue to the short arm of the translocation chromosome. The long arm was homologous to the longest acrocentric autosome. By C-banding, a single, small area of centromeric constitutive heterochromatin on the translocation chromosome indicated a single centromere. The translocation in Brown Swiss may be homologous to the 1/29 Robertsonian translocation in many other breeds of cattle. The translocation chromosome appeared to have segregated normally from its homologues. Aneuploid karyotypes were not observed. Analyses of breeding data failed to detect any decrease in fertility of translocation heterozygous females as compared to females of the normal karyotype. It would be premature to recommend a selection program against this translocation in Brown Swiss, but widespread use of sires heterozygous for the translocation should not be encouraged.


Blood antigen, serum protein, and milk protein gene frequencies and genetic interrelationships in Holstein cattle.

Gene frequencies at ten blood group loci, one serum protein locus, and four milk protein loci were determined for the Holstein-Friesian breed in the United States. The sample consisted of 8630 cows in 51 herds from 10 states. Because of the close linkage among casein subloci and the concimitant rarity of crossover-recombinant groups, casein gene combination or haplotype frequencies were determined also. As one means of comparison of systems, indices of homozygosity and number of effective alleles were calculated. These indices were proposed also to be useful tools for monitoring changes in genetic variability of breeds. Genotypes within codominant systems and phenotypes associated with paired-system combinations generally were not randomly occurring. Paired system phenotypes within the sire sample corresponded much more closely to expectations of randomness.


Incidence of clinical mastitis in a herd of Holstein cattle.

Incidence of clinical mastitis in 760 lactations of 331 Holstein cows was studied for effects of sire, parity, and month of calving. Incidence of clinical mastitis was higher in hind quarters than fore quarters. Positive correlations between number of cases in one quarter with number in other quarters and between proportion infected in one quarter and proportion infected in one quarter and proportion infected in others indicated that occurrence of mastitis in one quarter was associated with above average frequencies of mastitis in the other quarters of the same cow. Clinical mastitis increased with increase in lactation number up to fourth lactation.  Cows calving in January and February more frequently had mastitis than those calving in March and April.


Facility for exercising dairy cows.

That dairy cows housed in confinement might benefit if they were exercised during the dry period prompted development of a facility for exercising dairy cows. Two methhods of the exerciser, which consist of a circular lane and a driving mechanism powered by a variable speed electric motor, are described. A speed of 3.5 k, per h provides a steady but comfortable pace for pregnant dairy cows and heifers. Many animals refused to go at speeds greater than 5.5 km per h.


Food and economics.

A much-needed national food policy will encompass not only the recommendations of the nutritionist, but also those of the economist and the politician-generalist. Decisions and tradeoffs as to goals--as well as means of achieving them--must be made. Should elimination of malnutrition in the population be the overriding goal? Should freedom of choice be maximized? Should present food habits be accepted--or changed? The various objectives are not mutually compatible. For a viable policy to emerge, a blending of disciplines is necessary. The important thing is to ask the right questions of the right people--economic questions of the economist, nutritional questions of the nutritionist, and political questions of the politician. Then, the answers can be put together in a spirit of compromise rather than contentiousness.


Developing behaviorally anchored scales for rating dietitians' performance.

Behaviorally anchored rating scales were developed for dietitians, using a variant of the methodology of Smith and Kendall (3). Approximately thirty dietitians, representing administration, patient services, education, and community nutrition participated in the development. The job tenure of the participants ranged from one year to over twenty years. Seven dimensions were identified, and each was provided with a rating continuum containing eight or nine behavioral anchors. The dimensions and their definitions are listed, and one scale is presented.


Conventional vs. formulated foods in school lunches. I. Comparison of students' food and nutrient intakes.

School lunches containing all conventional foods were designed to provide either one-third or one-half the Recommended Dietary Allowances for elementary students. The same nutrient goals were used in planning meals containing some formulated foods. Goals and specified portion sizes for first through third graders were distinct from those for fourth through sixth graders, and data from the two groups were maintained separately. Nutrients in meals and plate waste were analyzed to compare nutrient intake among the treatments. Formulated meals weighed less than control and conventional meals, and students ate higher percentages of both the weight and the nutrients served in formulated meals. Nutrient intake was higher when half the allowances was served, but waste was also higher.


Development of an automated form generating system for menu item data.

At the University of Missouri-Columbia, an Automated Form Generating System consisting of six computer programs was developed to generate the master menu, production forecast and usage forms, production worksheet, food order report, and selective menus for patients. In the initial testing of the programs, this computer-assisted system showed a 30 per cent reduction in time requirred by dietitians and clerks when compared with the manual system. Some expected benefits include ease in making menu changes, substantial reduction in errors, automatic updating of documents, and reduction in storage space for forms because documents will be generated just prior to a production week. The system will be implemented at a later date.


Organizational decisions for food procurement in hospitals.

Effective food procurement is an important foodservice management function and has been established as a complex managerial process facilitated through organizational decision-making. Although the importance of decisions made by dietetic professionals responsible for food purchasing is currently increasing because of world and national concerns, the findings of this survey revealed a gap between theory and practice of food procurement. Some trends were revealed concerning the academic preparation for food procurement in the curricula of various educational programs. Entry-level competencies in buying food need to be defined. To educate students to be competent in food procurement decisions, effective planning and working relationships among administrative personnel in hospital foodservice operations and college and university educators are necessary. The twenty criteria for effective food purchasing and the associated specific practices provide a basic approach for such effective planning and working relationships.


Fortification of hamburger with calcium, vitamin A, and ascorbic acid.

Patties were prepared from ground beef--some with and some without calcium, vitamin A, or ascorbic acid, either singly or in various combinations. After cooking, the patties were mixed with buns and mayonnaise, ground, and then fed to weanling rats. No single added nutrient or combination of added nutrients seemed to affect flavor, texture, or cooking losses of fat or water. Fortification with calcium and ascorbic acid increased liver storage of vitamin A and decreased liver:body weight ratios.


Stability of ascorbic acid in orange juice exposed to light and air during storage.

Single-strength orange juice prepared from three nationally available brands of frozen orange juice concentrate was analyzed for ascorbic acid after exposure to light and air for varying periods up to eighteen days. For two of the brands, brown, foil-covered bottles offered no greater protection against ascorbic acid loss than clear glass bottles. Storage time appeared to be a greater contributor to ascorbic acid loss than type of container in two brands. One brand showed both the effect of storage time and type of container, since the juice stored in the brown bottle lost significantly less ascorbic acid after the same period than juice stored in the clear bottle.


Indirect fluorescence of primary and secondary myofibers in developing porcine muscle.

Cytochemical differentiation of two populations of developing skeletal myofibers has been demonstrated in fetal muscle with metachromatic fluorescence of ribonucleic acid and deoxyribonucleic acid by staining fresh frozed cryostat sections of developing porcine skeletal muscle with acridine orange (CL. 46005). Evidence is presented that supports the hypothesis that first-formed myofibers (primary myofibers) serve as a structural framework upon which myoblasts proliferate, fuse in linear sequence and give rise to a second population (secondary myofibers) of myofibers.


Human transfer factor prepared by dialysis, ultrafiltration and gel chromatography: biological activity in local transfer of skin sensitivity.

Human transfer factor (TF) was prepared by a variety of methods including dialysis using cellophane tubing, ultrafiltration through a membrane of known pore size. Sephadex G25 chromatography or combinations of some of these methods. In general the various preparations when injected locally into human skin gave greater delayed-type responses than antigen (PPD or Candida) alone. The combination of either vacuum dialysis, or ultrafiltration, with G-25 chromatography gave as good or better TF activity when compared with unchromatographed materials. Since ultrafiltration and concentration is rapid procedure and eliminates the need for freeze-drying, in contrast to vacuum dialysis against water, these results indicate that ultrafiltration and G-25 chromatography provide a convenient method for preparing large batches of relatively pure TF from leucocyte extracts.


Detection of circulating immune complexes in human sera by simplified assays with polyethylene glycol.

The search for circulating immune complexes by precipitation tests using polyethylene glycol (PEG) was performed on a series of normal and pathological sera. Various factors affecting PEG precipitation were studied. Immunoglobulins and complement factors percipitated by PEG (3.5%) were quantified and their significance was discussed in relation to serum levels. The PEG test was compared to labeled C1q binding test with a fairly good correlation. The direct evaluation of the amount of C4 precipitated with IgG by 3% PEG (C4 test) provided a simpler routine assay than the C1q binding test for detecting complement-fixing immune complexes. The direct PEG test and the C4 tests gave positive results in patients with diseases generally presumed to be associated with immune complexes including systemic lupus erythematosus, acute glomerulonephritis, bacterial sub-acute endocarditis and chronic acitve hepatitis. The demonstration of HBs antigen and antibody after acid dissociation of PEG precipitates from hepatitis B seronegative sera illustrated the fact that PEG does precipitate and thus concentrates circulating immune complexes.


Ultrastructural studies of nuclei and nucleoli in psoriatic epidermis, compared with those in normal epidermis.

Quantitative and qualitative differences of the nuclear and nucleolar structures in normal and psoriatic (involved and uninvolved) epidermis were revealed with the electron microscope. The size of nuclei and nucleoli was increased, the frequency of nuclear bodies was increased, and the relative amount of heterochromatin was decreased in involved skin compared to normal or uninvolved skin. There was no significant difference between normal skin and uninvolved skin of psoriasis. In addition to these changes, multiple fibrillar centers within nucleoli and small masses of chromatin in the epidermal nucleoplasm were observed very frequently in involved skin, while rarely or not at all in normal and uninvolved skin. These results suggest that involved psoriatic keratinocytes have increased metabolic activity and abnormal protein synthesis, having undergone gene derepression.


Decreased thymidine incorporation in circulating leukocytes after treatment of psoriasis with psoralen and long-wave ultraviolet light.

Tritiated thymidine incorporation, a measure of DNA synthesis, was studied in circulating leukocytes from patients with widespread psoriasis who were being treated with photochemotherapy using oral 8-methoxypsoralen (8-MOP) and high-intensity, long-wave ultraviolet light (UVA). Seven of 13 psoriasis patients treated with photochemotherapy demonstrated a significant (p less than 0.05) reduction in leukocyte incorporation of tritiated thymidine immediately after UVA in comparison to incorporation before UVA. None of 10 control subjects treated with UVA alone demonstrated such reduction in leukocyte tritiated thymidine incorporation. Photochemotherapy thus affects circulating blood cells in some patients with psoriasis in addition to its therapeutic effect on epidermal cells. Further investigations are needed to determine the reasons for the differences in susceptibility to inhibition of leukocyte DNA synthesis among patients and the possible long-term consequences of such inhibition.


The induction of immune tolerance in delayed contact sensitivity by the use of chemically related substances of low immunogenicity.

Immune tolerance in delayed contact sensitivity to pentadecylcatechol can be induced by a series of derivatives substituted in the 6 position of the ring. Some of these derivatives have the property of being very poor sensitizers and having very low dermal toxicity. Thus, sensitization and tolerance have different biologic mechanisms and are associated with different properties of these chemicals.


Induction of human skin sensitization to ethanol.

In predictive skin sensitization tests, 50% aqueous ethanol induced delayed allergic skin reactivity in 6 of 93 human volunteers. This was confirmed 2 months after the primary challenge by producing an allergic response in each of the 6 with a single 24-hr application of 50% aqueous ethanol. Three of these reactive subjects also responded to lower concentrations of ethanol in water, and 2 of them still showed allergic reactivity 18 months later. Possible allergens other than the ethanol itself (i.e., an impurity or an ethanol-soluble component of the patch) were eliminated. Responses of 1 subject to ethanol that had been purified by gas chromatography confirmed that ethanol alone was the sensitizer. This subject also reacted to other short-chain primary alcohols, a secondary alcohol, and acetaldehyde, suggesting an antigenic relationship among all of these compounds in the skin. One of the authors, who had become sensitized to acetaldehyde, showed a similar pattern of cross-reactivity. Previous infrequent reports of ethanol sensitivity have been limited to case studies and diagnostic patch testing. This work provides evidence that a test designed to predict skin-sensitizing potential will identify even a weak sensitizer such as ethanol. Aqueous ethanol solutions approaching 50% concentrations should be avoided as vehicles in human predictive sensitization testing.


In situ identification of lymphoreticular cells in benign and malignant infiltrates by membrane receptor sites.

Immunologic membrane markers have been utilized for the in situ identification of lymphoreticular cells within lymphoreticular tissues and in inflammatory and neoplastic lymphoreticular infiltrates of the skin. Frozen sections were layered with untreated sheep red blood cells (SRBC) for the detection of the E receptor on T cells, SRBC coated with 7S antibody for the detection of the Fc receptor (IgGEA) and SRBC coated with 19S antibody and complement (IgMEAC) for the detection of the C3 receptor. In normal lymphoreticular tissue, IgGEA selectively bound to areas colonized by macrophages, IgMEAC to B-dependent areas, whereas E showed no adherence. In chronic lymphatic leukemia, a predominant and selective adherence of IgMEAC was found in lymphoreticular organs which supports the B-cell nature of the neoplastic cells; in leukemic reticuloendotheliosis a selective IgMEAC adherence was observed and this favors a monocytoid origin of the hairy cells. Adherence of any indicator cell was practically absent from skin infiltrates of mycosis fungoides, which represents an additional piece of evidence for the T-cell nature of this disease process. In psoriasis, a considerable binding of IgGEA was observed, whereas IgMEAC did not adhere at all. This result indicates that the lymphocytic part of the infiltrate in this disease consists mainly of T cells.


Dimorphism of Pityrosporum orbiculare in a defined culture medium.

Dimorphism of Pityrosporum orbiculare was induced in an artificial culture medium which consisted of 0.05 M glycine in 0.03-0.06 M ammonium phosphate buffer (pH 5.6), salts, glucose, and Tween-80. Of the strains tested, 78 to 85% produced hyphal elements from up to 8.5% of yeast cells. Morphologic features of P. orbiculare in culture were examined by scanning electron microscopy and compared with the appearance of fungus from clinical lesions of tinea versicolor. Morphologically, the fungi obtained in vivo and in vitro were identical.


Early host cell-Molluscum contagiosum virus interactions.

Basal and suprabasal layers of human epidermis infected with the poxvirus Molluscum contagiosum have been examined with the technique of serial sectioning. Phagocytic vacuoles, formerly not observed in human epidermis, were found exclusively in the basal region. They did not fuse with other virus-containing vacuoles or with lysosomes to form digestive vacuoles. Various stages of uncoating, preceding ejection of the virus core into the cytoplasm, were observed in the virus-containing vacuole. Clusters of cores were commonly found close to or even associated with centriolar structures. Their possible interference with mitosis is discussed in relation to alterations observed in the plasma membrane. It is assumed that excision of gap junction elements precedes the induction of mitosis.


A comparison of the glycosaminoglycans of weight-bearing and non-weight-bearing human dermis.

Twenty-six samples of human skin from three non-weight-bearing and 27 samples from three weight-bearing sites were obtained at autopsy from 11 subjects varying in age from 4 to 91 years. The dermis was idssected free of other tissues and after proteolytic digestion, the glycosaminoglycans were isolated quantitatively by ion-exchange chromatography. Analysis of the preparations showed that there was significantly more glycosaminoglycan in the dermis from weight-bearing sites (p less than 0.001). The major glycosaminoglycans were hyaluronic acid and dermatan sulfate with chondroitin sulfate as a minor component, but the relative proportions of these components were not significantly different in the two sets of samples (p greater than 0.1). It is suggested that the quantity of glycosaminoglycan in the dermis is influenced by the functional requirements of the dermis at that site.


Follow-up studies in patients with right bundle branch block and left anterior hemiblock: significance of H-V interval.

Thirty-eight patients with right bundle branch block and left anterior hemiblock who had undergone His bundle recordings were prospectively followed for development of heart block. Twenty-five were male and thirteen female. Their ages ranged from 35 to 92 with an average age of 74 years. The mean follow-up period was 28.5 months (range 2-52 months). The H-V interval was 55 msec or less in 17 patients (Group A), between 56 and 75 msec in 13 (Group B), and more than 75 msec in eight (Group C). Ten patients (26%) died during the follow-up period. Of these, three were in Group A (mortality rate 8% per year), three in Group B 10% per year), and four in Group C (21% per year). None of the deaths in Groups A and B were sudden, while one possible sudden death occurred in Group C. Mode of death in one patient each from Groups B and C remained unknown. Heart block was not seen in any patient from Group A and B, while two patients from Group C progressed to complete heart block during the follow-up period. It is concluded that patients with right bundle branch block and left anterior hemiblock with prolonged H-V intervals tend to show a higher mortality when compared to those with normal H-V intervals. A definite risk of heart block exists in patients with H-V intervals of more than 75 msec.


Conduction disturbances in patients with mitral valve prolapse.

The electrocardiograms (ECGs) of 55 patients with mitral valve prolapse were analyzed. Twelve of these demonstrated conduction abnormalities. Five patients had first degree heart block. Three had the classic ECG changes of the Wolff-Parkinson-White syndrome. Two patients had left anterior hemiblock, one of whom also had incomplete right bundle branch block. One patient had episodes of advanced atrioventricular block and periods of sinus arrest, which resulted in recurrent syncopal attacks. One patient had incomplete left bundle branch block. The presence of conduction disturbances points to the diffuse nature of the myocardial abnormality in mitral valve prolapse.


Polarcardiographic criteria for infarction evaluated by angiocardiography.

Out of a series of 1000 consecutive patients studied by angiocardiography, two groups were selected on the basis of the probability of having suffered myocardial infarction: an infarction group of 324 cases, and a non-infarction group of 112 cases. Members of the infarction group had complete occlusion of at least one major coronary artery; members of the non-infarction group had less than 50% occlusion in all coronary arteries, normal hemodynamics, and negative histories of infarction. The diagnostic performances of the independently-read 12-lead electrocardiograms (ECGs) and the polarcardiograms (PCGs) were evaluated in terms of sensitivity, specificity, validity and risk ratio. The performance of the ECG was significantly lower than the PCG. Corresponding figures were 56%, 99%, 67%, and 1.67 for the ECG, and 75%, 91%, 80%, and 2.23 for the PCG. There is a 99.9% probability that the PCG can detect at least 21% more cases of infarction than the ECG can.


Diagnosis of high posterior infarction: experimental study through the use of body surface isopotential maps.

This investigation was designed to diagnose high posterior infarction easily through the use of body surface isopotential maps. High posterior infarction was experimentally caused by ligation of the branch of the circumflex artery of the canine left coronary artery. Each dog had a series of maps recorded before and four weeks after experimentally induced myocardial infarction. The common features of maps in high posterior infarction are: 1) The positive area expanded into the dorsal surface so slowly that, in the middle stage of the ventricular depolarization, a large area of the dorsal surface was still covered by the negative area. 2) from the middle to the late stage, the absolute value of the minimum significantly decreased and absolute potential of the dorsal surface also significantly decreased. In conclusion, we propose that it is possibile to diagnose high posterior infarction through the use of maps.


Genesis of body surface potential distribution in right bundle branch block.

In order to investigate the specific sites of conduction block in the three types (I, II, III) of right bundle branch block (RBBB) classified by body surface isopotential maps, the simulation of ventricular propagation process and mathematically reconstructed maps were used. Four assumptions were introduced from the results of clinical observations and animal experiments. The maps reconstructed from two of these assumptions, in which the conduction block was placed on the main stem of the right bundle branch, showed two different patterns at late stages of excitation, and these two kinds of map resembled Types I and II in clinical maps, respectively. The maps reconstructed from the other two assumptions, in which the site of the conduction block was located mainly in the Purkinje system of the right ventricular free wall, resembled Type I at the late stage of excitation in one of two assumptions and agreed with Type III through all stages of excitation in other case. Based on the above results, it is speculated that the differences of ranges and degrees of conduction block ascribed to abnormal activation in the Purkinje system of the right ventricular free wall are responsible for the genesis of clinical RBBB map patterns.


Electrophysiologic properties of intravenous oxprenolol in man.

Electrophysiologic modifications produced by intravenous administration of 0.1 mg/Kg Oxprenolol were studied in 16 subjects with estimated normal impulse formation and conduction. Significant effects were sinus bradycardia, mild increase of sino-atrial conduction time, depression of intranodal conduction and prolongation of A-V node refractory periods. Sinus node recovery time and atrial refractory periods were unchanged. Infranodal conduction and the refractory periods of the His-Purkinje system, as well as of the bundle branches, were unchanged. These effects are compared with those observed after intravenous Propranolol and Prindolol.


Multiple conduction defects with markedly prolonged ventricular depolarization in cardiomyopathy.

Electrophysiologic investigation of a diffuse and bizarre conduction abnormality in a patient with cardiomyopathy revealed profound slowing of the impulse in the right ventricular outflow region. The resultant 220 msec QRS duration is the longest reported in right bundle branch block.


Cardiac abnormalities in a female patient with hypogonadotropic hypogonadism with anosmia.

Hypogonadotropic hypogonadism with anosmia was found in a 38 year old female. Cardiac abnormalities were manifested by a second degree heart block as well as a conduction delay below the AV node. Striking T wave inversions developed in the absence of any symptoms. These changes probably reflect a central nervous system defect.


2:1 left bundle branch block in acute bacterial endocarditis with septal abscess.

This report presents an unusual type of conduction disturbance of the left bundle branch which appeared in a patient with acute bacterial endocarditis of the aortic valve in which the septum was also invaded. This is the first case found in which the partial left bundle branch block (LBBB) was of acute inflammatory origin. Other previously reported cases of this functional delay of the left branch were secondary to arteriosclerotic heart disease.


Recurrent paroxysmal supraventricular tachycardia: a complication of ventricular pacing in a patient with occult Wolff-Parkinson-White syndrome.

A 60 year old man suffering from syncope believed to be due to the sick sinus syndrome was treated with a permanent demand ventricular pacemaker. This led to almost continous bouts of paroxysmal supraventricular tachycardia (SVT) over the ensuing two years, mistakenly believed to be part of the sick sinus syndrome. Careful study showed that this man had a type A Wolff-Parkinson-White accessory atrioventricular connection which consistently conducted retrogradely, but only rarely antegradely, during applications of carotid sinus massage. Episodes of SVT were repeatedly induced whenever ventricular-paced impulses captured the atria retrogradely. All episodes of SVT stopped when the ventricular pacemaker was removed. Following insertion of an atrial pacemaker, the patient had no episodes of SVT or syncope over a nineteen month follow-up period. This case illustrates the care required in selecting a proper site for protective pacing in patients who suffer from paroxysmal SVT.


Stimulus similarity as a determinant of Pavlovian conditioning.

Three experiments carried out second-order Pavlovian conditioning using either similar or dissimilar first- and second-order stimuli. Experiments 1 and 2 used rat subjects in a conditioned suppression and an appetitive conditioning preparation, respectively. Experiment 3 used pigeons in an auto-shaping procedure. All three experiments were designed to identify the effects of similarity upon conditioning as distinct from its effects upon sensitization or stimulus generalization. Each experiment found superior conditioning when similar stimuli were paired. Several theoretical implications of these findings are discussed.


Bisection of temporal intervals.

Eight rats were trained to make one response if a signal was shorter than a criterion duration and a different response if the signal was longer than the criterion. When exposed to intermediate durations, the rats bisected the interval at the geometric mean and the difference limen divided by the geometric mean was a constant. The rats learned new temporal discriminations more easily when the response maintained its relative, rather than its absolute, meaning. These data were interpreted in terms of a model of an internal clock that included a clock, a criterion, and a response rule.


Rolyadenylic acid [poly(A)] sequences associated with measles virus intracellular ribonucleic acid (RNA) species.

The kinetics of 3H-uridine incorporation into measles-infected Vero cells demonstrated that maximum virus-specific RNA synthesis occurred between 16 and 20 h after infection. Sedimentation analysis on sucrose gradients revealed the presence of four species of RNA having sedimentation coefficients 4S, 12 to 26S, 28 to 36S and 50S. Annealing studies showed that RNA sedimenting in the 12 to 36S regions was 100% complementary in base sequence to nucleocapsid 50S RNA, and at least 96% of the 50S genomic RNA was transcribed during virus replication. Polynucleotide binding experiments ane ribonuclease treatment indicated that poly(A) sequences were associated with the intracellular 12 to 26S, 28 to 36S and 50S RNAs. Denaturation of intracellular 50S RNA followed by sucrose gadient centrifugation demonstrated that this was a mixture of genomic 50S and heterogeneous RNAs which sedimented at 4 to 40S. The genomic RNA did not contain poly(A) sequences, and these are presumably associated with the heterogeneously sedimenting RNAs. The size of poly(A) sequences present on the 12 to 36S RNAs was estimated to be in the range of 70 to 140 nucleotides. Treatment of the 12 to 36S RNAs and their poly(A) sequences with polynucleotide phosphorylase indicated that the poly(A) was located on the 3' end of the RNAs, but that under the experimental conditions used this was protected by the secondary structure of the molecules.


Metabolism of interferon: hepatic clearance of native and desialylated interferon.

The effect of removal of sialic acid on the survival of rabbit serum and urinary interferon (IF) has been investigated in isolate, perfused rabbit liver preparations. In contrast with native IF, which may be already partially desialylated. IF freed of 80 to 90% of its sialic acid was rapidly cleared from the perfusate of normal livers, or livers pre-treated with actinomycin D. The results suggest that the mechanism of IF catabolism by the liver is similar to that reported for several other circulating glycoproteins.


Complementation of adeno-associated satellite virus (AAV) by temperature-sensitive mutants of adenovirus type 31.

Temperature-sensitive (ts) matants of human adenovirus type 31 were able to complement adeno-associated satellite virus (AAV) antigen production in both HEK and KB cells at both permissive and non-permissive temperatures. However, mutant ts 94, an adenovirus 31 mutant which produces apparently normal amounts of structural protein and DNA but is defective in maturation, was significantly inhibited in its ability to potentiate AAV infectivity at the non-permissive temperature. Normal AAV DNA and adenovirus DNA were isolated from co-infections with AAV and mutant ts 94 at the non-permissive temperature. We suggest that an adenovirus-coded maturation function common to both adenovirus and AAV maturation is defective in the ts 94 system.


Isatin-beta-thiosemicarbazone causes premature cessation of vaccinia virus-induced late post-replicative polypeptide synthesis.

The effects of isatin-beta-thiosemicarbazone on vaccinia virus-induced polypeptide synthesis has been examined by polyacrylamide gel electrophoresis and autoradiography. The synthesis of pre-replicative and early post-replicative polypeptides proceeded normally in the presence of the drug; the onset of late post-replicative polypeptide synthesis occurred with normal timing under these conditions but the rate of synthesis of all virus polypeptides then declined rapidly.


The effect of cycloheximide on the replication of measles virus.

Treatment of measles virus-infected cells with cycloheximide results in a three-fold increase of 3H-uridine incorporation into the 12 to 36S mRNA species and in the inhibition of genomic 50S RNA synthesis. Consistent with these observations was the finding of a build-up of polyribosomes but an absence of nucleocapsids in the infected cells. These results suggest that measles virus RNA replication, but not transcription, is dependent upon active protein synthesis.


The relation of auditory, visual, and auditory-visual matching to reading performance of Israeli children.

Six same-different matching tests, both verbal and nonverbal, in three modalities, along with a set of reading tests, were administered to 120 Israeli children in second, third, and fourth grade. The main effect of all S variables, except sex (grade, socioeconomic level, and ability) were significant, as were the test factors of modality (visual, auditory, cross-modal) and form (verbal-nonverbal), but interactions between S and test factors were small. Multiple regression analysis revealed that overall matching test scores accounted for 35% of the variance in reading scores, although the additional contribution of specific subtests was negligible. Performance on the visual-visual tests was virtually perfect. Auditory-auditory matches were more difficult than auditory-visual matches with nonverbal stimuli, while the reverse was true with verbal stimuli.


The factorial structure of adolescent responses to a fear survey schedule.

The Wolpe-Lazarus Fear Survey Schedule was administered to a sample of 553 girls and 559 boys ranging in age between 12 and 18 years, and their responses were factor analyzed. The most prominent factors to emerge were those loading on items in the Social stimuli and Tissue damage categories. An analysis of frequency of response to schedule items indicated that this prominence was not a problem of item distribution in the schedule. The findings are discussed in relation to theoretical accounts of fears, and the author concludes that no one particular theoretical position can adequately account for the major findings.


Effects of estrogen on prolactin and tuberoinfundibular dopaminergic neurons.

Serum prolactin concentrations and dopamine turnover in the striatum and median eminence were studied in male rats after the administration of estradiol benzoate. The alpha-methyltyrosine-induced reduction of dopamine concentrations in these brain regions was used to evaluate relative rates of turnover. Steady state dopamine concentrations in the median eminence and striatum were not altered by 1, 3 or 5 days of estradiol treatment. However, 3 or 5 days of estradiol administration enhanced dopamine turnover in the median eminence but not in the striatum. Estradiol treatment failed to alter dopamine turnover in the median eminence of hypophysectomized rats. Estradiol increased serum prolactin concentrations at all of the times examined. Although alpha-methyltyrosine also increased serum prolactin, this increase was further enhanced in estradiol-treated rats. The increased prolactin response to alpha-methyltyrosine and increased dopamine turnover in the median eminence of estradiol-treated rats suggests that tuberoinfundibular dopaminergic neurons may be part of a hormonal-neuronal negative feedback loop which functions to regulate prolactin secretion.


Reaction of foetal tissues to intra-uterine subcutaneous injection of barium sulphate.

Subpannicular injection of sterile barium sulphate in newborn rabbits produces an acute inflammatory response which is clearly seen at 24 hr and is well established at 48 hr. The processes of repair has begun by the 4th day with the appearance of proliferating capillaries and fibroblasts. In rabbit foetuses both vascular and cellular components of the acute inflammatory response are more prominent and appear earlier than in newborn rabbits, being well developed within 24 hr. The process of repair also begins earlier, proliferation of capillaries and fibroblasts being already prominent by 48 hr. By the 4th day the lesion is compact, less cellular and relatively avascular.


Morphological study of lesions induced by snake venoms (Naja naja and Agkistrodon piscivorus) in the lung and cremaster vessels of rats.

The morphological effects of two snake venoms, N. naja and A. piscivorus, and of the Direct Lytic Factor and Phospholipase-A, compounds purified from N. naja crude venom, were investigated on lung and cremaster vessels of rats. The microcirculation of the rat reacts to these two venoms differently: N. naja produces congestion, haemolysis and increased vascular permeability, whereas A. piscivorus causes these alterations, plus haemorrhage and thrombosis. Direct Lytic Factor elicits reponses similar to the N. naja venom but Phopholipase-A has no effect on the vessels. Phospholipase-A does not seem to potentiate the effects of Direct Lytic Factor on the cremaster vessels. The colloidal carbon technique showed that intrathroacic administration of N. naja venom results in a generalised permeability increase of pleural and subpleural capillaries and A. piscivorus injections provoke only carbon retention in capillaries at sites of localised haemorrhage. In cremasters treated with N. naja venom the carbon blackened the venules predominantly but in cremasters in which A. piscivorus venom was administered the carbon particles labelled both venules and capillaries. There was evidence that the main vascular action of the venoms is local and not systemic, that the permeability factors involved in these lesions are different and that the cremaster vessels are much more sensitive to these snake venoms than the pulmonary vessels. Electron-microscopic studies showed mesothelial and epithelial lesions in lungs and an early inflammatory reaction in the cremaster vessels with both venoms. Erythrocyte fragmentation was a constant feature in all vessels. Endothelial degeneration and capillary disintegration in lung and cremaster vessels were observed in animals treated with A. piscivorus venom.


Adult Niemann-Pick disease--a case report.

A case is presented of the rare adult form of Niemann-Pick disease occurring in a Cantonese. The diagnosis was verified by biochemical analysis of the affected tissues.


Humoral antibodies against spermatopoietic cells associated with mediastinal germinoma.

A case of mediastinal germinoma in a 21-yr-old male is reported. The neoplasm exhibited a morphology similar to that of a testicular seminoma and was heavily infiltrated by lymphocytes and plasma cells. Germinal centres were also present in lymph follicles. Humoral antibodies (IgG), specific for spermatopoietic tissue, were present in the patient's serum. Antibody titres declined after surgical removal of the germinoma over a period of 2 yr, and antibody is no longer detectable at the present time. The implications of this observation are discussed with regard to defence mechanisms of the tumour host, and to the histogenesis of these rare neoplasms.


The fine structure of human macrophage granules in sarcoidosis.

Human macrophages in the lesion of sarcoidosis contain cytoplasmic granules with an ordered paracrystalline substructure. Similar though less ordered granules are seen in human macrophages elsewhere. These appearances as well as the general ultrastructure of the cell are compatible with the view that these macrophages have a secretory function.


Transformation in the presence of metals of lymphocytes from patients with total joint prostheses.

A lymphocyte transformation test has been used to study metal sensitivity in patients receiving total joint replacements. In the case of nickel, very significant levels of transformation were obtained with lymphocytes from patients with positive patch tests for this metal. In the case of patch test negative recipients of total joint prosthesis an elevated index of transformation was found in seven out of 15. Transformation in the presence of cobalt was less striking and there was evidence of toxicity of the metal in doses used to obtain stimulation. In eight out of 12 patch test positive individuals a significant level of transformation was obtained. Two of the seven patients carrying joint prostheses who wwere also patch test negative gave an elevated index of transformation. The clinical implications of these results are discussed.


The physiologically relevant information regarding systemic blood pressure encoded in the carotid sinus baroreceptor discharge pattern.

1. The objective was to find out what kind of informatioon regarding systemic blood pressure is transduced by baroreceptors in vivo and how this information is coded in the receptor discharge. 2. Carotid sinus pressure, e.c.g., and receptor action potentials were recorded for fifty-two single fibre carotid sinus receptors found in twenty decerebrated unanaesthetized cats. 3. The inflation and gradual deflation of an intraaortic catheter tip balloon manipulated the blood pressure in the carotid sinus in a way as to define the full in vivo stimulus-response curve for each receptor. 4. Correlation coefficients were computed between stimulus and response variables for several points on the response curve of each receptor and for every possible combination of stimulus and response variables defined. 5. Stimulus variables were (a) systolic, (b) diastolic,, (c) mean, (d) pulse pressures and (e) peak positive dP/dt. Response variables were (a) average discharge rat, (b) peak instantaneous frequency, and (c) average burst frequency. 6. For every fibre in the sample only the correlations between systolic, diastolic and mean pressures vs. average discharge rate were consistently high and positive. All other correlations were numerically low and/or negative. 7. It was concluded that in vivo baroreceptors signal mainly pressure level (systolic, diastolic or mean) as opposed to pulse pressure or dP/dt, and that the average discharge rate is their best index of information content.


The physiological rhythms of subjects living on a day of abnormal length.

1. Fourteen subjects, singly or in groups, have been observed while living on a 21 hr day for 8 or 16 experimental 'days' and fifteen other subjects similarly on a 27 hr day. 2. Rhythmic components of body temperature and excretion of various urinary constituents were calculated. 3. On a 21 hr day, for most components and most subjects, two periods were present, one of 21 hr and one of around or somewhat over 24 hr. 4. On a 27 hr day two periods were less often present and a larger number of observed rhythms could be satisfactorily described by a single period, usually between 23 and 28 hr. 5. In subjects spending a second week on a 21 hr day the circadian component was no less prominent than during the first week. 6. When, after life on a 21 hr day, subjects were deprived of knowledge of time, there was evidence that the 21 hr component did not persist. 7. The results are interpreted as evidence of the continuing existance of an influence with a period of around 24 hr, simultaneously rhythmic influences resulting from the subjects' habits. On a 27 hr day there was sometimes evidence of entrainment, yielding an intermediate period. 8. An attempt is made to compare the relative potency of the exogenous and of the persistent circadian influences on the several variables.


Eimeria galateai sp. n. from the Paradise Kingfisher, and Eimeria duncani sp. n. from the Sacred Kingfisher in Papua New Guinea.

Eimeria galateai sp. n. from the paradise kingfisher (Tanysiptera galatea Gray) and Eimeria duncani sp. n. from the sacred kingfisher (Halcyon sancta Vigors & Horsfield) have been described from Papua New Guinea. Four of 11 paradise kingfishers were infected with E. galateai oocysts, measuring 13 (11-16) X 9 (8-11) micron. The oocysts were ovoid with nipple-like protrusion at one pole. Micropyle and polar granule were absent, while oocyst residuum (5 X 4 micron) was present. Sporocysts, measuring 5 (4-6) X 2 micron, were elongate-ovoid, and had a distinct convex Stieda body; the sporocyst residuum was absent. Two of 9 sacred kingfishers were infected with ovoid-truncated, 22 (19-25) X 16 (12-18) micron oocysts of E. duncani. Polar granule (5 X 2) was present in the oocysts, but there was no micropyle or oocyst residuum. Sporocysts were ovoid, measuring 9 (8-10) X 5 (4-6) micron, with a prominent Stieda body, and granular sporocyst residuum. Eimeria galateai and E. duncani are the first species of this genus to be described from birds of the order Coraciiformes.


Coccidia (Protozoa: Eimeridae) of the herbivorous mole-rat, Spalax leucodon Nordmann.

A survey of 96 herbivorous mole-rats, Spalax leucodon Nordmann, in Lalahan district of Ankara, Turkey, revealed the presence of 6 new species of coccidia. The species found, the shape and average length-width dimensions (in micrometers) of their oocysts, and the numbers of animals from which they were isolated were as follow: Eimeria lalahanensis sp. n., subspherical to ellipsoidal (20.3 X 12.5), from 10; Eimeria tuzdili sp. n., spherical to subspherical with radially striated oocyst wall, (20.5 X 16.5), from 3; Eimeria spalacis sp. n., ellipsoidal to cylindrical (16.3 X 9.8), from 90; Eimeria elliptica sp. n., ellipsoidal (14.3 X 10.1), from 8; Eimeria turkmenica sp. n., ellipsoidal (11.3 X 8.9), from 1; Isospora anatolicum sp. n., spherical (7.6 X 5.1), from 1 animal. The overall infection rate for 96 examined herbivorous mole-rats was 100%.


Five new species of Henneguya (Protozoa: Myxosporida) from ictalurid fishes.

Four new species of Henneguya (myxosporidan) are described from Ictalurus punctatus Rafinesque (channel catfish). They are as follows: Henneguya postexilis sp. n. and Henneguya longicauda sp. n. from the gills; Henneguya adiposa sp. n. from the adipose fin; and Henneguya diversis sp. n. from the liver, kidney, connective tissue of muscles and fins, and tumor-like external growths. Henneguya pellis sp. n. is described from the dermis of Ictalurus furcatus (LeSueur) (blue catfish). The development stages of all described species are discussed. Henneguya exilis Kudo was found on the gills of one I. punctatus; notes on its spore characteristics are presented.


Light and electron microscopic observations on encystment of Acanthamoeba palestinensis, Reich.

The ultrastructural changes occurring during encystment of acanthamoeba palestinensis have been investigated. The cyst wall consists of endocyst and exocyst, both having the same fine structure. At irregular intervals in the cyst wall ostioles occupied by opercula are present. The nuclear membrane forms bulb-shaped projections and releases vesicles bounded by double membranes into the cytoplasm. Dense nucleolus-like bodies of different sizes and variable numbers are found in the nucleus of every cyst. The importance of the cyst structure as a taxonomic criterion is discussed.


Effect of chloramphenicol on bacterial endosymbiotes in a strain of Amoeba proteus.

The effect of chloramphenicol (CAP) on the bacterial endosymbiotes of a strain of Amoeba proteus was studied by growing the symbiotic amebae in media containing 0.5-1.6 mg/ml CAP for up to 4 weeks. Treatments with CAP caused such ultrastructural changes as expansion of the nuclear zone and deformation of symbiotes. The CAP treatment also damaged the mitochondria, e.g. disappearance of central and protrusion of peripheral cristae. Number of bacteria per ameba decreased to less than 10% of control in CAP-containing media, but no viable amebae became completely free of symbiotes. The results supported previous studies that amebae were dependent on endosymbiotes.


Piroplasms from Taiwanese rodents.

Piroplasms from naturally infected bandicoot rats, Bandicota indica (Hodgson), and spiny rats, Rattus coxinga (Swinhoe), from Taiwan caused parasitemias in a wide range of laboratory animals after syringe passage in laboratory rats. Bandicoot and spiny rats could be infected with heterologous piroplasms, but neither species was susceptible to infection with the Gray strain of Babesia microti (França). Nevertheless, the Taiwan piroplasms are probably a geographic strain of B. microti.