Diffusion-limited ion flow through pores. If the permeability of an ion channel is high, the overall transport becomes ultimately limited by the rate with which ions from the aqueous phase arrive at the mouth of the channel. In this study a theoretical treatment of diffusion-limited ion transport through pores is given which takes both concentration and electric potential gradients into account as driving forces for the flow of ions. The current-voltage characteristic of a diffusion-limited channel is found to be strongly saturating. Addition of an impermeable elesults from a reduction in the electric field strength near the mouth of the channel. At low voltages the permeability P of the pore is obtaines as 1/P =(1/Pi)+(1/P'c)+(1/P''c) where Pi is the intrinsic permeability and P'c, P''c are the left-hand and right-hand convergence permeabilities, respectivelyl which are proportional to the aqueous diffusion coefficient of the permeable ion and to the effective capture radius of the pore.


Serum glycoprotein-type sequence of monosaccharides in membrane glycoproteins of Semliki Forest virus. Semliki Forest virus was grown in BHK-21 cells and labelled in vivo with radioactive monosaccharides. The virus was disrupted with sodium dodecyl sulphate and the polypeptides were hydrolyzed with pronase. A mixture of type A glycopeptides (for nomenclature, see Johnson and Clamp (1971) Biochem. J. 123, 739-745) of the membrane glycoproteins E1 and E3 was isolated by gel filtration and subjected to sequential degradation with exo-glycosidases. The reduction in the apparent molecular weight and the cleavage of radioactive monosaccharides were monitored with gel filtration. The results suggest that the type A oligosaccharides have similar average structures and contain at the non-reducing terminus 3.4 mol of alpha-D-sialic acid and 0.7 mol of alpha-L-focose, folloled by 3.1 mol of beta-D-galactose, 4.2 mol of N-acetyl-beta-D-glucosamine, 0.7-1.5 mol of alpha-D-mannose, 0.5 mol of beta-D-mannose and 0.6-2.2 mol of N-acetyl-beta-D-glucosamine attached to 1.0 mol of N-acetylglucosamine resistant to N-acetyl-beta-D-glucosaminidase. This innermost monosaccharide unit, therefore, appears to be attached to the peptide. The peptides attached to this N-acetyl-glucosamine had an apparent molecular weight of 720+/-100. We propose the following average structure, compatible with most of our data, for the type A glycopeptides of Semliki Forest virus:.


The interaction of an anionic photoreactive probe with the anion transport system of the human red blood cell. N-(4-azido-2-nitrophenyl)-2-aminoethyl[35S]sulfonate is employed as a photoreactive probe for the anion transport system in the human erythrocyte. In the dark and at 37 degrees C the probe penetrates the membrane via a pathway sensitive to specific inhibitors of anion permeability. It reversibly inhibits sulfate and chloride fluxes but the inhibition is reduced by higher concentrations of sulfate. Upon photolysis to produce a reactive nitrene (at 0 degrees C to minimize penetration), the probe inhibition of anion permeability. Under appropriate conditions the degree of inhibition after photoactivation (irreversible) is almost the same as that in the dark (reversible). The binding sites for the radioactive probe are largely found in proteins of 95 000 apparent molecular weight (band 3). After pronase treatment of the labelled cells, most of the probe is found in a 65 000 molecular weight segment derived from the 95 000 molecular weight protein. In this respect the photoreactive probe resembles another potent irreversible inhibitor of anion transport, 4, 4'-diisothiocyano-2, 2' stilbene disulfonate. In fact, most of the binding sites for each probe are common to both. Thus, in the dark, the azido derivative protects the anion system from inhibition by DIDS and substantially reduces the binding of DIDS to band 3 protein. Conversely, pretreatment with DIDS substantially reduces the binding of the photoreactive probe to the same protein. The fact that an apparent substrate for the anion permeation system competes for binding sites with a specific non-penetrating inhibitor of anion permeability suggests that the inhibitory and transport sites may be closely related and implicates the 95 000 molecular weight protein as the element of the anion transport system which contains the substrate binding site.


Error introduced by small reflection coefficients in permeability constansts obtained by hemolysis. In 1933, M.H. Jacobs (J. Cell. Comp. Physiol. 4, 161-183) developed the theoretical basis for calculating permeability constants of nonelectryolytes passively penetrating erythrocytes from experimentally determined hemolysis times in isotonic solution of penetrating solute. This derivation had assumed that the reflection coefficient sigma=1, whereas, usually 0 less than sigma less than 1, By comparison of Jacobs' original derivation with the equations revised to include sigma, it is shown that: (see article) where r=(k1/k2), the apparent ratio of solute permeability constant (k1) to osmotic volume flow constant of water (k2) as determined by the Jacobs approach; and ro=(omega/Lpcs), the ratio of the true permeability constant (omega) to the osmotic flow calculated from the product of the pressure-filtration coefficient (Lp) and the concentration gradient (cs). The correct ratio may be expressed as a function of the apparent ratio: (see article) For large or small values of r, simpler approximations may be used: r greater than 1, ro approximately r r less than 1, ro approximately sigma4r These provide less than 20% error if sigma greater than 0.4 and r greater than 10 or r less than 0.1. Corrections for sigma less than 1 are applied to the classical (k1/k2) permeability constants of ethylene glycol, glycerol and propanol for bovine eryghtocytes. The sigma values for monoacetin and diacetin are predicted to be 0.7 and 0.6, respectively, on the basis of the deviation of their (k1/k2) constants from the expected relationship to partition coefficient and vapor pressure.


Effect of bilayer curvature on vibrational Raman spectroscopic behavior of phospholipid-water assemblies. In order to clarify the effect of bilayer curvature upon phospholipid conformation, vibrational Raman spectra were recorded for dipalmitoyl and dimyristroyl phosphatidylcholine in the gel state for both multilayer and single-wall vesicle assemblies. An intensity comparison, based upon a nonperturbing internal standard, between the two classes of bilayrer systems reflected a decrease in peak height intensity for the observed hydrocarbon chain transitions in the single shell vesicle form. No intensity change between bilayer form was detected, however, for the two observed head group modes. Trends in the peak height intensity rations for the 1100 cm-1 carbon-carbon stretching vibrations indicated an increase in hydrocarbon chain transgauche isomerization for the vesicle in comparison to the multilayer arrangements. The sensitivity of the methylene carbon-hydrogen stretching modes to interchain interactions was demonstrated by comparisons of the intensity patterns in the 2900 cm-1 region to the intensity characteristics of the carbon-carbon stretching region for polycrystalline, multilayer and vesicle materials. Examination of various carbon-carbon stretching mode intensity ratios for cholesterol doped dipalmitoyl phosphatidylcholine bilayers indicated that while 25 mol% cholesterol increased the transgauche acyl chain isomerization in multilayers, no comparable effect was observed for the vesicle forms. In contrast, the methylene twisting/methylene deformation intensity ratios for the cholesterol containing systems suggested that some further type of interchain perturbation occurs in the vesicle aggregations.


The preference of cholesterol for phosphatidylcholine in mixed phosphatidylcholine-phosphatidylethanolamine bilayers. The following phosphatidylethanolamines were studied by differential scanning calorimetry: 1,2-dipalmitoleoyl-, 1,2-dioleoyl-, 1,2-dilauroyl-, 1,2-dielaidyl-, 1,2-dimyristoyl- and 1,2-dipalmitoyl-sn-glycero-3-phosphoryl-ethanolamine. The saturated and trans-unsaturated species underwent thermotropic phase transitions at temperatures about 20-30 degrees C higher than the corresponding phosphatidylcholines but the enthalpy changes were nearly identical. The transition temperatures for the cis-unsaturated species were about the same as those of the corresponding phosphatidylcholines but here the enthalpy change was markedly decreased as compared with the phosphatidylcholines. Freeze-fracture electron microscopy revealed phase changes from a lamellar to a hexagonal phase for 1,2-dipalmitoleoyl- and 1,2-dioleoyl-sn-glycero-phosphorylethanolamine at 20 and 0 degrees C respectively. At these temperatures no transitions were apparent in the calorimeter scan. Incorporation of increasing amounts of cholesterol into phosphatidylethanol-amine bilayers gradually decreased the enthalpy changes of the phase transition in the same manner as was demonstrated before for phosphatidylcholine/cholesterol mixtures. This was studied both for 1,2-dipalmitoleoyl- and 1,2-dimyristoyl-sn-glycerophosphorylethanolamine. In an equimolar mixture of 1,2-dioleoyl- and 1,2-dipalmitoylphosphoryl-ethanolamine, which showed phase separation, cholesterol preferentially decreased the transition of the lowest melting component. In equimolar mixtures of phosphatidylethanolamines and phosphatidylcholines, which showed phase separation, cholesterol preferentially abolished the transition of the phosphatidylcholine component present. This occurred both in experiments where the phosphatidylcholine was the lowest melting and where it was the highest melting component present in the mixture. These experiments strongly suggest that in phosphatidylcholine-phosphatidylethanolamine mixtures at temperatures where both components are in the liquid-crystalline state cholesterol is preferently associated with the phosphatidylcholine component in the mixture.


Chemical modification by trinitrobenzenesulfonate of a lipid and proteins of intracytoplasmic membranes isolated from Chromatium vinosum and Azotobacter vinelandii. 1. The structure of intracytoplasmic membranes of a photosynthetic bacterium chromatium vinosum and a nitrogen-fixing bacterium Azotobacter vinelandii was studied by chemical modification of amino groups of phosphatidylethanolamine and proteins with trinitrobenzenesulfonate. 2. Almost all the constituents of intracytoplasmic membranes of C. vinosum were solubilized in a mixture of chloroform, methanol and trichloroacetic acid. One-third of proteins in the intracytoplasmic membranes of C. vinosum was found solubilized in a mixture of chloroform and methanol. By using a column chromatography with Sephadex LH-20 in organic solvents, the unmodified as well as the trinitrophenylated proteins and also the trinitrophenylated phosphatidylethanolamine were separated from the other colored substances. 3. In the chemical modification of the intracytoplasmic membrane preparations, 30% of phosphatidylethanolamine and 15% of protein amino groups in C. vinosum and 45% of phosphatidylethanolamine and 20% of protein amino groups in A. vinelandii were estimated to be exposed to the aqueous phase. In the single-layered liposomes composed of phosphatidylethanolamine and phosphatidylglycerol with a ratio of 2:1, 40% of phosphatidylethanolamine were estimated to be exposed to the aqueous phase.


Fusion and protein-mediated phospholipid exchange studied with single bilayer phosphatidylcholine vesicles of different density. A novel method has been developed for the study of phospholipid exchange and fusion of phospholipid vesicles. Two homogeneous populations of single bilayer phosphatidylcholine vesicles of similar size but markedly different density have been prepared. "Dense" vesicles were made from brominated dioleoyl phosphatidylcholine. "Light" vesicles were prepared from dioleoyl phosphatidylcholine. The two populations were easily separated by density gradient centrifugation. Phosphatidylcholine exchange protein from beef liver was used to promote lecithin exchange between the vesicle populations. Only the lecithin of the external monolayers of the vesicles was available for exchange by exchange protein, implying that flip-flop of vesicle phosphatidylcholine did not take place at a detectable frequency. No spontaneous intervesicle phosphatidylcholine exchange was observed. However, the dense and light vesicles did spontaneously fuse, over several hours, to produce particles of hybrid density.


Lectin-receptor interactions in liposomes. II. Interaction of wheat germ agglutinin with phosphatidylcholine liposomes containing incorporated monosialoganglioside. Bovine brain gangliosides incorporated into phospholipid liposomes provide receptors for wheat germ agglutinin. Purified monosialogangliosides were mixed with egg phosphatidylcholine, and unilamellar liposomes were generated. Addition of wheat germ agglutinin induced the liposomes to fuse, and gel filtration analysis revealed that the lectin was incorporated into the fused liposomes. The fusion process was studied by following the changes in the 90 degrees light scattering. Increasing the proportion of the monosialoganglioside in the liposomes was found to increase both the extent of the lectin-induced liposome fusion and the rate of the reaction; below a threshold of approx. 5 mol%, the process was extremely slow. The increase in light scattering could be prevented by the addition of the hapten inhibitor, N-acetyl-D-glucosamine (1 mM). Addition of the inhibitor, subsequent to the lectin, caused a partial decrease in light scattering due to the dissociation of unfused vesicle aggregates. Electron microscopic examination revealed that the ganglioside-containing liposomes were vesicles, 244 +/- 25 A (S.D.) in diameter. Upon addition of wheat germ agglutinin, the vesicles appeared to fuse to form larger vesicles, corresponding to dimers and trimers of the initial vesicles. Inhibition studies with a variety of monosaccharides indicated that the sialic acid moieties present in the ganglioside acted as the lectin-receptor sites. This was confirmed by the observation that wheat germ agglutinin did not interact with phosphatidylcholine vesicles containing desialyated ganglioside.


The lipid composition of plasma membrane subfractions originating from the three major functional domains of the rat hepatocyte cell surface. 1. The neutral and phospholipid compositions of three rat liver plasma membrane subfractions originating predominantly from the three major functional domains of the hepatocyte viz the blood sinusoidal, contiguous and bile canalicular fractions, were determined. 2. The sinusoidal and canalicular plasma membrane subfractions, both of which were vesicular, contained a higher lipid to protein weight ratio than the contiguous plasma membrane subfraction that consisted of membrane strips, junctional complexes and some larger vesicles. The three plasma membrane subfractions contained a similar neutral lipid to phospholipid ratio. The highest unesterified cholesterol content was associated with the canalicular plasma membrane subfraction. 3. The phospholipid profiles of the three subfractions were generally similar. However, the canalicular plasma membrane subfraction contained a higher proportion of sphingomyelin than the other subfractions. 4. Correlations between the neutral and phospholipid composition of the subfractions and membrane integrity and function are discussed, especially with respect to a possible role of lipids in governing the resilience of the canalicular plasma membrane to the action of bile salts.


Mechanism of ion transport through lipid bilayer-membranes mediated by peptide cyclo-(D-Val-L-Pro-L-Val-D-Pro). The cyclic dodecapeptide PV, cyclo-(D-Val-L-Pro-L-Val-D-Pro)3, a structural analogue of the ion-carrier valinomycin, increases the cation permeability of lipid bilayer membranes. This paper reports the results of two types of relaxation experiments, namely relaxation of the membrane current after a voltage jump and decay of the membrane voltage after a charge pulse in lipid bilayer membranes exposed to PV. From the relaxation data, the rate constant for the translocation of the ion carrier complex across the membrane, as well as the partition coefficient of the complex between water and membrane solution interface were computed and found to be about one order of magnitude less than the comparable values for valinomycin (Val). Furthermore, the dependence of the initial membrane conductivity on ion concentration was used to evaluate the equilibrium constant, K, of complexation between PV and some monovalent cations in water. The values of K yield the following selectivity sequence of PV: Na+ less than NH4+ less than K+ less than Cs+ less than Rb+. These and earlier results are consistent with the idea that PV  promotes cation movement across membranes by the solution complexation mechanism which involves complexation between ion and carrier in the aqueous phase and transport of the carrier across the membrane. In the particular form of the solution complexation mechanism operating here, the PV present in the PV-cation complex carrying charge across the membrane derives from the side from which the current is flowing (cis-mechanism). As shown previously, valinomycin, in contrast to PV, acts by an interfacial complexation mechanism in which the Val in the Val-cation complex derives from the side toward which current is flowing (trans-mechanism). The comparison of the kinetic properties of these two closely related compounds yields interesting insights into the relationship between chemical structure and function of ion carriers.


Transport kinetics of hydrophobic ions in lipid bilayer membranes. Charge-pulse relaxation studies. A modified version of the charge-pulse relaxation technique with improved time resolution was applied to the study of transport kinetics of hydrophobic ions (tetraphenylborate, dipicrylamine) through lipid bilayer membranes. Besides a better time resolution the charge-pulse method has the additional advantage that the perturbation of the membrane can be kept small (voltage amplitudes between 1 and 10 mV). The results of the analysis support the model proposed earlier, according to which the overall transport takes place in three consecutive steps, adsorption of the ion from water to the interface, translocation to the opposite interface, and desorption into the aqueous phase. The translocation rate constant ki and the partition coefficient gamma of the hydrophobic ion between water and the membrane were measured for lecithins with different mono-unsaturated fatty acid residues. Increasing the chain length of the fatty acid from C16 to C24 resulted in a decrease of ki by a factor of about 9 in the case of tetraphenylborate and by a factor of about 17 in the case of dipicrylamine.


Voltage-induce capacitance relaxation of lipid bilayer membranes. Effects of membrane composition. The specific capacity of black lipid membranes of different phospholipids dissolved in n-alkanes was investigated. The hydrocarbon thickness of these membranes, as calculated from the electric capacity with a dielectric constant of 2.1, was in most cases close to 5 nm. It was found that the specific capacity is not constant with time after blackening. It shows a linear time dependence characteristic for each lipid/solvent system. The influence of a transmembrane potential on the capacity of the membranes was measured. It was shown that the extent of the capacity change, obtained 10 s after applying a voltage, was strongly dependent on the lipid composition as well as the solvent content of the membranes. The capacity change of the membranes seems to be caused mainly by a thickness change and not by an area increase of the membranes.


Structural changes in bilayer membranes by multivalent ions. Formation of microlenses, a reduction in thickness, and a change in mechanical properties have been observed when Ca2+ or other multivalent ions are added to bathing solutions of Mueller-Rudin membranes of acidic phospholipids. The observations are interpreted as an extrusion of residual solvent from the hydrocarbon core of the bilayer due to changes in packing imposed by the reduction of electrostatic repulsion of the head groups.


Active potassium transport in reticulocytes of high-K+ and low-K+ sheep. The kinetics of active K+ transport were studied in immature red blood cells cells from high-K+ and low-K+ sheep particulary with respect to the effects of varying intracellular K+ concentration, [K]i. Comparison was made with active transport, or pump, activity in mature high-K+ and low-K+ red cells. Reticulocytes from both types of sheep had much higher maximal active K+ influxes than did mature cells. In both types of reticulocytes, and in mature high-K+ cells as well, the pump was relatively insensitive to increasing [K]i. In contrast, intracellular K+ markedly inhibited the pump in mature low-K+ cells. Active K+ transport in low-K+ reticulocytes, however, as in mature low-K+ cells, is stimulated by specific isoimmune anti-L serum. Therefore the K+ pumps of high-K+ and low-K+ reticulocytes have similar kinetic properties. Maturation of the red cells, involving inactivation of most of the pump activity in both cell types, results in mature high-K+ and low-K+ cells with K+ pumps of very different kinetic characteristics.


A specific decrease of the fluorescence depolarization of perylene in muscle membranes from mice with muscular dystrophy. The microviscosity of erythrocyte membranes and muscle microsomes from age matched 6-week old control mice REJ 129 Dy/Dy, and mice with muscular dystrophy REJ 129 DY/DY has been estimated by measuring the fluorescence depolarization of perylene. There was no difference between the erythrocyte membranes. The muscle microsomes from dystrophic animals had about 20% lower values than the controls. The temperature dependence indicated that a transition occurs in both sets of muscle microsomes, but the transition temperature was lower in the dystrophic microsomes. Cholesterol, phospholipid and triglyceride analyses of the membranes showed no difference between the erythrocyte membranes. The largest difference in the muscle microsomes was a two-fold increase in cholesterol level found in the dystrophic microsomes. No simple correlation could be made between the lipid analysis and the microviscosity measurements. Since the change in microviscosity is found in membranes isolated from the tissue primarily affected by the dy gene, we suggest that the change in microviscosity may be important in the development of the disease.


Voltage-clamp experiments on oxidized cholesterol membranes modified with excitability-inducing material and comparison with a model. The conductance of oxidized cholesterol membranes modified with excitability-inducing material was observed in membranes containing either single conductance channels or 100-1000 channels. Membranes containing single channels have several conductance states for each voltage polarity, and the current through membranes containing manychannels decays with at least two, and probably three, time constants following a step change in voltage (voltage-clamp). The time constants differ by about an order of magnitude. The multi-state behavior seems more pronounced in membranes made from highly oxidized cholesterol. Although a given conductance state could occur at either positive or negative voltages, each state was much more frequent at one polarity or the other. The most frequently observed single-channel conductance states in 0.1 M NaCl are about 0.3, 0.1, 0.03, and 0.0 n-1 for negative voltages and 0.25, 0.05, 0.03, and 0.0 n-1 for positive voltages. The current following a voltage clamp decays to a quasi-steady state within 1 min for positive voltages and 1-15 min for negative voltages. When the holding voltage is --20 mV, the decay constants and quasi-steady state conductances as functions of clamping voltage are reasonably well described by either a three-state model of the conductance or a two-state model applied independently at negative and positive voltages. However, for high voltages, the quasi-steady state does not appear to approach a state in which all the channels are in a low conductance state.


Purification and partial characterization of two glycoproteins in bovine peripheral nerve myelin membrane. Two major glycoproteins of bovine peripheral nerve myelin were isolated from the acid-insoluble residue of the myelin by a procedure involving delipidation with chloroform/methanol (2:1, v/v) and chromatography on Sephadex G-200 column with a buffer containing sodium dodecyl sulfate. The separation patterns of the proteins on the gel were affected considerably by the dodecyl sulfate concentration in the elution buffer. At above 2% dodecyl sulfate concentration in the elution buffer, the glycoproteins, could be separated clearly on the gel and were purified. The purified proteins, the BR protein (mol. wt. 28 000) and the PAS-II protein (mol. wt. 13 000), were homogeneous on dodecyl sulfate-polyacrylamide gel electrophoresis. The NH2- terminal amino acids of the BR and the PAS-II proteins were isoleucine and methionine, respectively. The BR protein contained glucosamine, mannose, galactose, fucose and sialic acids and the PAS-II protein contained glucosamine, mannose, galactose, fucose and glucose. Neither the BR protein nor the PAS-II were a glycosylated derivative of a basic protein of bovine peripheral nerve myelin, a deduction based on the results of amino acid analysis. The two major glycoproteins were observed commonly in the peripheral nerve myelin of cows, pigs, rabbits and guinea pigs, using dodecyl sulfate-polyacrylamide gel electrophoresis.


Fractionation of human erythrocyte membranes. Presence of the nucleoside transport complex in an insoluble residue. (1) Human erythrocyte membranes, when dialysed against water at pH 9.5, were partly solubilized, losing 80% of the membrane proteins and 65% of the membrane lipids. Sodium dodecyl sulphate gel electrophoresis of the particulate material revealed selective removal of proteins from the membrane. (2) The lipid-rich particulate material remaining retained the ability to bind specifically the nucleoside transport inhibitor, nitrobenzylthioinosine, previously shown to bind selectively to the nucleoside transport mechanism of whole erythrocytes and erythrocyte ghosts.


Modulation of membrane drug permeability in Chinese hamster ovary cells. The kinetics of colchicine uptake into Chinese hamster ovary cells have been investigated and found to be consistent with an unmediated diffusion mode. A variety of compounds such as local anesthetics and non-ionic detergents as well as drugs such as vinblastine, vincristine, daunomycin and actinomycin D potentiate the rate of colchicine uptake into these cells and into colchicine resistant mutants. In all cases, higher concentrations of these compounds were required to stimulate colchicine uptake in the colchicine resistant mutants than in the cells of the parental line. This stimulation was observed also in the uptake of puromycin, a structurally and functionally different drug. These stimulatory agents did not, however, cause the cells to become nonspecifically leaky since the uptake of 2-deoxy-D-glucose was unaffected. In addition, the activation energy of colchicine uptake was unaltered in the presence of stimulating agents, implying that they were not causing colchicine to enter the cells via a different mechanism. The results are compatible with the view that these compounds are membrane-active, and are able to stimulate an increased rate of unmediated diffusion of colchicine into the cells. It appears that a mechanism for the regulation of passive permeability is modified in the resistant mutants.


Testing the simple carrier using irreversible inhibitors. 1. We analyse the kinetics of irreversible inhibition of the simple carrier. We consider how the rate of inactivation is influenced by the concentrations of permeant on the two sides of the membrane. 2. We consider various kinetic schemes for the simple carrier and show that these are all indistinguishable kinetically, using steady-state transport or inactivation data. We point out the advantages of using the simplest kinetic scheme and the possible pitfalls of using more complicated schemes, including the conventional carrier model. 3. We show that in the absence of information on the transport properties of the system, irreversible inhibition data are ambiguous. Taken together with transport data, however, inactivation data provide new tests for the applicability of the simple carrier. 4. The new tests show that for the simple carrier model to be applicable, the substrate dependencies of transport and of inactivation must be identical in comparable experimental situations. Further, the maximal rates of transport and of stimulation (or inhibition) of inactivation must obey a simple relationship, which we derive. We illustrate the use of these tests with published data on the glucose and choline transport systems of the human red blood cell.


Determination of Triton X-100 binding to membrane proteins by polyacrylamide gel electrophoresis. The molecular weight of proteins in protein-detergent complexes can be determined from ultracentrifugation experiments if the amount of bound detergent is known. A new sensitive method to measure the binding of the nonionic detergent Triton X-100 to proteins has been developed. For the membrane proteins studied, less than 50 mug of protein was required to achieve an accuracy of 10% in the determination of the detergent-protein weight ratio. The proteins were equilibrated with the detergent by electrophoresis into polyacrylamide gels containing radioactively labelled Triton X-100. The gels were then sliced and the amount of bound detergent calculated from the increase in radio-activity in the slices containing the protein zone. The amounts of protein were determined by amino acid analysis of identical protein zones cut from gels running parallel.


Molybdate and tungstate transfer by rat ileum. Competitive inhibition by sulphate. For both MoO42- and WO42- the maximum rate of uptake by the small intestine of the rat (studied in vitro using the everted sac technique) occurs in the lower ileum. Kinetic constants, derived by a least squares procedure, are compared with those previously obtained for SO42- transport. For both V and Ka, SO42- greater than MoO42- greater than WO42-, with only small differences between sacs IV and V. Mutual inhibition of MoO42- and WO42- transport and inhibition of both by SO42- are competitive processes. This is shown by the generally good agreement between Ka values and derived Ki values and by V values in the presence and absence of the inhibiting species. The three ions SO42-, MoO42- and WO42- are probably transferred across the intestine by a common carrier system. Implications for the sulphate-molybdenum interaction in molybdosis are discussed.


Stoichiometry of hemolysis by the polyene antibiotic lucensomycin. The stoichiometry of hemolysis by the polyene antibiotic lucensomycin was investigated. It appears that hemolysis occurs only when a relatively high fraction (probably between 15 and 40%) of the cholesterol sites in the erythrocyte membrane have combined with the polyene. Also in phospholipid-cholesterol vesicles the increase of permeability requires occupancy of 40-50% of the existing cholesterol sites. As for the possible cooperative effect in the hemolytic process, it is probable that several (at least 9-10) lucensomycin-cholesterol adducts must interact on each side of the membrane to form an aqueous channel; the distribution of these adducts in the erythrocyte membrane occurs, however, apparently at random.


Selective effects of nonionic detergent and salt solutions in dissolving nuclear envelope protein.?20U. Protein has been selectively extracted from isolated chicken erythrocyte nuclear envelope by (1) dilute MgCl2/Triton X-100 followed by (2) concentrated MgCl2/Triton X-100 solutions. Certain proteins appear to be selectively dissolved in the first solvent and may occur in the nuclear envelope primarily as lipoproteins. Among the proteins insoluble in the low MgCl2/Triton X-100 wash, as well as in 500 mM MgCl2 without Triton previously used in the preparation of the envelope fraction, the quantitatively major polypeptides dissolve in a combination of high MgCl2 and Triton X-100. Further, much of this dissolved protein precipitates when the MgCl2 concentration is lowered by dialysis. The insolubility of these proteins appears to result from a combination of ionic and hydrophobic interactions and may explain the resistance of nuclei to various manipulative procedures including nonionic detergent washes. The procedures described provide a route for gently and selectively dissolving representative proteins from the nuclear envelope lipoprotein matrix and from the envelope "residual" protein.


[Inducible cephalosporinases from Proteus morganii]. It has been shown that two Proteus morganii strains produce an inducible cephalosporinase. No significative difference was shown between them: they present the same isoelectric poit: 8,3 and very similar kinetic constants. The parameter tau, proportional to the half life of antibiotic at low concentration, in presence of beta lactamase, shows that cefamandole is the most stable cephalosporin studied.


Structural studies on RNA from Bombyx mori L. I. Nucleoside composition of enriched tRNA species from the posterior silkgland purified by coutercurrent distribution. A large scale fractionation of tRNA from the posterior silkgland of the silkworm Bombyx mori L. by countercurrent distribution is described. One single 1,500 transfer distribution carried out with Phosphate buffer-Fromamide-Isopropanol (PFI) solvent system yields highly enriched isoaccepting species with increasing mobility order: tRNA1Gly, tRNA1-2Ala, tRNATyr, tRNA2Gly, tRNA1Ser and tRNA2Ser with 75%, 70%, 90%, 60%, 60%, and 90% purities respectively. Nucleosides fingerprint analysis of each iso-tRNA species confirms the anticodon structures previously suggested for tRNA2Ala (IGC), tRNA2bGly (U-CC) (U-CC) and tRNA2bSer (IGA). Twenty two minor nucleosides, three of them with unknown structure, have been detected. They are: m5C in tRNA1Gly, m1I in all tRNAAla species, polar A and U called X in tRNATyr, polar U derivative in tRNAGly2, mt6A in tRNASer1 and i6A tRNA2Ser. Both tRNASer species have m3C and ac7C. We do not detect Q, Y and thiol derivatives. The elution characteristics of silkgland tRNA species may be expressed in a semilogarithmic diagram where log K (K is the partition coefficient) is related to the base ratio A/Y) and the coding properties. The distribution pattern of silkgland tRNAs has been compared with that of Yeast and Rat liver tRNAs fractionated by countercurrent distribution with the PFI and PMB (Potassium phosphate buffer, 2-methoxy ethanol, 2-butoxy ethanol) solvent systems.


Mouse brain uptake and metabolism of stearic acid. After injection, labelled stearic acid is transported directly into the brain and incorporated into brain lipids without prior oxydation to acetate and resynthesis of fatty acids. Contamination by blood can be excluded. (The preparation contains all subcellular fraction except cytosol). The labelled stearic acid taken up is partly metabolized in the brain either by elongation or by degradation and in situ resynthesis of fatty acids. The activity in oleic acid and mono-unsaturated chains is hardly detectable. The labelled acids are incorporated into lipids or subcellular particles following characteristic kinetics, which show a diminution by 24 hours. When analysing the evolution of each lipid, it is shown that this profile is followed by phospholipids, but not by cerebrosides and free fatty acids. The formers are still increasing up to 50 hours, the latters are stable (suggesting a physical binding between membranes and free fatty acids). Thus nutrition is an important parameter for the synthesis of brain membranes as far as exogenous saturated fatty acids are needed.


[Kinetic and structural characteristics of succinate dehydrogenase components reacting with natural and artificial electron acceptors]. A new catalitic activity of soluble succinate dehydrogenase, i.e. the reduction of low (20-200 muM) concentration of ferricyanide in the presence of succinate is described. The apparent Km value for the acceptor is about 200 muM. The turnover numbers of the enzyme measured in this reaction, with PMS as an electron acceptor and in the system reconstituted from soluble enzyme and alkali-treated submitochondrial particles (succinate oxidase) are found to be almost the same. The new succinate. ferricyanide reductase activity is very sensitive to oxygen, high (3 mM) ferricyanide concentration and mercaptide-forming agents. When the enzyme is stored under aerobic conditions the loss of this activity occurs according to the first-order kinetics with the same rate constants as the reconstitutive activity decreases. The rate constants both for ferricyanide reductase and reconstitution decay do not depend on pH within the range of 6,5--7,5 (k = 8.10(-2) min-1) and increase dramatically at pH 8,5 (K = 4.10(-1) MIN-1). When these two activities are lost after oxygen exposure the PMS-reductase fall down to about 50% of its original activity. The new ferricyanide reductase is found only in the soluble preparation of the enzyme succinate: cytochrome c reductase, succinate dehydrogenase of submitochondrial particles and reconstituted succinate oxidase do not interact with low concentrations of ferricyanide. The treatment of the enzyme after inactivation by oxygen exposure with sulfide ion--iron--mercaptoethanol mixture followed by Sephadex filtration completely restores the original reconstitutive, ferricyanide and PMS reductase activities. The hypothesis is suggested that succinate dehydrogenase contains at least two red-ox centers reacting with electron acceptors. The first one is located in hydrophylic environment (mitochondrial matrix) being accessible for high concentrations of ferricyanide. The second one (iron--sulfur complex, Hipip-type) is responsible for ferricyanide reductase activity described, being located intramembraneously and involved in the electron transfer between dehydrogenase and the rest of the respiratory chain.


[Mechanism of the phosphoglucomutase reaction]. The points of bonds ruptures in the course of phosphoglucomutase reaction were studied by two methods: incorporation of 18O and kinetic isotope effect with the use of double label (14C and 3H). The method of 18O determination by means of activation analysis is described. Analysis of data leads to the conclusion that phosphoryl, but not phosphate group, moves in the course of the reaction. Possible mechanisms of phosphoglutcomutase reaction are discussed.


[Protein and RNA synthesis during the transition of potato tuber meristematic tissue from rest to growth]. The rate of RNA and protein synthesis in resting and growing meristem tissues from isolated tuber growth points under their incubation with labelled precursors is studied. It is found that the label incorporation into RNA is low during deep resting, protein synthesis being rather intensive. Both RNA and protein synthesis is intensified with the beginning of growth period. More homogenous in their metabolic rate set of RNAs is found in resting tissues in contrast with growing ones. The data on fractionation of total RNA preparation on MAK column and in sucrose density gradient showed that synthesis of high molecular weight RNA with metabolic period of 4 hours is stimulated in growing meristems. It is possible that synthesis of certain fractions of RNA and protein is one of regulation mechanisms for resting in meristem tissues.


[Glycine and alanine synthesis enzymes in the tissues of the silkworm during its development]. The activity of enzymes of glycine and alanine synthesis (glutamate-pyruvate aminotransferase, aspartate-beta-decarboxylase, threonine aldolase, serine hydroxymethyltransferase, alanine-glyoxylate aminotransferase, aspartate aminotransferase) is studied in haemolymph, fat body, fibroin and sericine divisions of silk gland of silkworm Bombyx mori at terminal period of larva development. Alanine-glyoxylate aminotransferase activity in fibroin division of silk gland (34,6 mu mole of glycine/mg of protein/min-10(-3)), alanine aminotransferase--in sericine division (36,0 mu mole of alanine/mg of protein/min-10(-3)) aspartate aminotransferase 27,3 mu mole of glutamic acid/mg of protein/min-10(-3)) and alanine aminotransferase (35,8 mu mole of alanine/mg of protein/min-10(-3)) on fat body. The ratio of alanine-glyoxylate aminotransferase/glutamate-pyruvate aminotransferase activities in posterior division of silk gland is near to glycine/alanine ratio in silk fibroin. The character of the enzymes activity in silkworm tissues correlates with the silk formation rate.


[Interaction of 4-aminobutyrate-transaminase from swine kidneys with 5'- and 6'-methyl derivatives of pyridoxal-5'-phosphate]. The study of interaction of 4-aminobutyrate transaminase with 5'- 6'-methyl derivates of PLP demonstrated that only the former was capable of forming a catalytically active holoenzyme possessing 0.37 activity of the native holoenzyme and a low affinity substrates. This compound interacts with the apoenzyme at a slower rate than does PLP; it has a reduced affinity towards apotransaminase (Km = 1.10(-4) M) and is replaced from the active site by native coenzyme. The other analog of pyridoxal-5'-phosphate forms a catalytically inactive complex with the apoenzyme; the other analog is not replaced from the active center by native coenzyme and non-competitively inhibits the reconstruction of apotransaminase (Ki = 2.10(-5) M).


[Stability of the polyribosomes of Blakeslea trispora Thaxter during normal translation, its stimulation and inhibition]. beta-Ionone is found to stimulate considerably carotinoids synthesis in Blakeslea trispora. The stabilization of carotene-synthesizing ability of B. trispora in the presence of beta-ionine under prolonged incubation time is observed. Stabilization of polyribosomes in the presence of beta-ionine is observed when studying polyribosome content in B. trispora. A hypothesis is expressed on the existence of biochemical "receptors" as a linkage between synthesized protein and destroying mRNA.


[Effect of hydrocortisone and actinomycin D at different time of the day on incorporation of labeled precursor into the nuclear proteins of intact rat liver]. Variations in the incorporation of labelled precursor into nuclear rat liver proteins during a day in normal conditions and in the presence of hydrocortisone and actinomycin D are studied. Synthesis of nuclear proteins is found to have a fluctuating character. Hydrocortisone and actinomycin D, injected at different day time, induce changes in the character of nuclear proteins synthesis , which are more pronounced for histones. The effect of the hormone and the antibiotic is tightly bound with the functional state of liver cells at given moment.


[Physico-chemical properties of a protein-inhibitor of serine proteinases from the potato]. Serine proteases inhibitor with pl-7.3, isolated from potatoe tubers by isoelectric focusing procedure as described previously (V.V. Mosolov et al., Bioorganic Chem., 1, 1449, 1975), was homogeneous under ultracentrifugation, having sedimentation coefficient S20,w 2.8S. Its molecular weight, investigated by sedimentation equilibrium and gel filtration through Sephadex G-100, was found to be 32500 and 31500 respectively. The Stokes radius R and frictional ratio f/fo were found to be 24 A and 1.14. The molecular weight of the inhibitor as determined by sodium dodecylsulfate polyacrylamide electrophoresis was twice as low as determined in ultracentrifuge and by gel filtration procedure. It is suggested that the inhibitor is dimer and consists of two protomers of equal molecular weight.


[Effect of polyene antibiotics on isolated dog kidney nuclei]. Effect of amphotericin B and nistation on caryoplasmic proteins, nuclear membrane-bound chromatin (DNPm) and soluble DNP (DNP0) from dog kidney isolated nuclei is studied in vitro. The yield of caryoplasmic proteins from nuclei is found to be increased during incubation with amphotericin B and nistatin, the content of some fraction in caryoplasmic proteins being decreased while their qualitative composition being unchanged. It is found that the treatment of nuclei with amphotericin B contributes the association of DNP particles with nuclear membrane and the increase of protein/DNA and RNA/DNA ratios in DNPm. No such effects are observed in the presence of nistatin. Both antibiotics do not affect protein/DNA and RNA/DNA ratios in DNP0 fraction. Polyene antibiotics are shown to change considerably the composition of acid soluble proteins in DNP0 and DNPm and in non-histone proteins in DNP0, and not to affect the content of lipids and their fatty acid composition in DNPm. The data obtained are of certain value for explanation of the mechanism of toxic effect of polyene antibiotics on animal cells.


[DNA degradation during standard alkaline of thermal denaturation]. Essential degradation 8 DNA (up to 10 per cent) with liberation of acid-soluble fragments takes place on the standard alkaline (0,01 M sodium phosphate, pH 12, 60 degrees, 15 min) or thermal (0.06 M sodium phosphate buffer, pH 6.8, 102 degrees C, 15 min) denaturation. This degradation is more or less selective: fraction of low molecular weight fragments, isolated by hydroxyapatite cromatography and eluted by 0.06 M sodium phosphate buffer, pH 6.8 is rich in adenine and thymine and contains about 2 times less 5-methylcytosine than the total wheat germ DNA. The degree of degradation of DNA on thermal denaturation is higher than on alkaline degradation. Therefore while studying reassociation of various DNA, one and the same standard method of DNA denaturation should be used. Besides, both the level of DNA degradation and the nature of the resulting products (fragments) should be taken into account.


[Search for a protein kinase specific for treponin T]. A specific to troponin T (as compared to f1 histone) proteinkinase is isolated using affinity chromatography on Sepharose 4B with covalently bound dephosphorylated troponin T. Homogeneous proteinkinase has a molecular weight of 175.000 and phosphorylates troponin T at a rate exceeding that of histone phosphorylation, while crude proteinkinase preparation has troponin T phosphorylation rate 15--20 times as low as that of histone phosphorylation. The enzyme isolated is capable of phosphorylating troponin T, but not troponin I, in the whole troponin complex.


Human EEG spectra before and during cannabis hallucinations. EEG correlates of subjective experiences induced by delta9-trans-tetrahydrocannabinol (THC) and EEG correlates of individual disposition to such experiences were investigated. Twelve normal volunteers took 200 mug/kg THC orally. The subjects were asked to signal subjective experiences. The EEG was analyzed (period analysis) before and repeatedly after THC injestion, during resting, attention, eye closure, visual hallucinations, and body image disturbances. EEG frequency spectra differed significantly between resting and visual hallucinations and body image disturbances. The differences included slower alpha and more theta during THC experiences, reminiscent of initial drowsiness EEG, and of some results in schizophrenia. The differences between spectra during visual hallucinations and during body image disturbances indicate different functional brain states. Subjects with a high tendency to cannabinol induced experiences exhibited resting spectra before and after THC with higher modal alpha frequences (reminiscent of subjects with high neuroticism scores) than subjects with a low tendency.


Short-term effects of naltrexone in 155 heroin ex-addicts. The narcotic antagonist naltrexone was administered for periods of up to 8 months to a total of 155 patients at a dose of 40-200 mg per day. The antagonistic effect of naltrexone was tested by injections of heroin. Eighty milligrams of natrexone was effective for 48hr. The antagonistic effect decreased at 72 hr after the administration of 120-200 mg of naltrexone. Laboratory tests indicated no signs of toxicity. Naltrexone may elicit an increase in blood pressure and opigastric pain. Neither of these side effects appear clinically important. No signs of dependence on naltrexone were detected. These results suggest that naltrexone may be useful for clinical treatment of opiate dependence.


Carotid-vertebral artery blood transit time: results in healthy subjects and patients with schizophrenia and brain disease. Estimation of blood transit time in the neck and the extension of this into the head was made by a method of electrical impedance, over a wide age range, in 159 healthy subjects, 160 schizophrenic patients, and 199 patients with organic brain disease. In each case, the distance between the electrocardiogram and the next succeeding pulse-volume impedance wave was measured and averaged over 30 serial wave forms. Chronological age proved a significant variable, transit times lengthening progressively with the age, but only for measurements extending into the head. Sex and hemispheric laterality played no significant role. With age held constant, mean transit times into the head were significantly prolonged in both groups of patients as compared with controls. No significant differences were found however between means of psychiatric andneurological patients. It is suggested that these results reinforce the organic etiology of schizophrenia.


Erythrocyte ITP pyrophosphohydrolase deficiency in a psychiatric population. A higher incidence of erythrocyte ITP pyrophosphohydrolase deficiency was found among a psychiatric population when compared to a nonpsychiatric group of subjects. Although the incidence was highest among schizophrenics, particularly those diagnosed as paranoid, the majority of the patients did not show such deficiency. The parallelisms between ITP pyrophosphohydrolase and hypoxanthine-guanine phosphoribosyl transferase deficiencies, both characterized by the lack of availability of IMP, and correlated to behavioral disorders, suggest that irregularities of hypoxanthine nucleotides may be implicated in abnormal mental processes.


The effect of voluntary control of heart rate deceleration on skin conductance level: an example of response fractionation. Twenty-four undergraduate male volunteers participated in an experimental designed to assess the effect of voluntary control of heart rate deceleration on skin conductance level. One group of subjects received heart rate feedback training and a second group performed a tracking task. Because heart-rate feedback was presented via a visual display, the tracking task group was included to control for display monitoring influences on heart rate. Results demonstrated feedback mediated acquisition of learned control of heart rate slowing. More importantly, the heart-rate slowing performance was accompanied by increases in skin conductance level. This 'fractionation' of physiological responding suggests the presence of a physiological response pattern which may counter initial attempts to produce greater magnitude slowing effects.


Further studies of physiological concomitants of hypnosis: skin temperature, heart rate and skin resistance. Forehead skin temperature, heart rate and palmar skin resistance were recorded during passive hypnosis and compared with corresponding data obtained during the resting awake condition in a group of highly hypnotizable subjects experienced in self-hypnosis. Similar physiological measures were also monitored during experimental periods when subjects were experiencing suggested environmental conditions of cold and heat in hypnosis as compared with imagining the stress conditions. The data from these subjects were also compared with those obtained from a randomly selected group of people who were low in waking suggestibility and had never been hypnotized. The results indicate that the differences in mean physiological parameters were greatest between the two subject groups, although some noteable differences were also apparent between hypnosis and the awake condition within the experimental group.


Relationship between basal body temperature and body activity in a human following a bilateral oophorectomy. Continuous measures of arm movement activity and basal body temperature were obtained on one female over a total of 149 days, beginning 3 months subsequent to this female's bilateral oophorectomy. Prior to the oophorectomy the subject had been menstruating regularly, and earlier data on this subject had indicated a low-order positive correlation between temperature and activity. The present results found no such correlation, lending support to the notion that at least one index of bodily activity is related to ovarian functioning.


Characteristics of a neural clock regulating perception and psychomotor performance in man. A biological clock regulating some psychomotor aspects of perception has been adduced from the results of the repetitive administration of a maximal speed tapping test of 15 sec per hand per min for a total of 20 min. 412 longitudinal timed tests from 8 healthy control subjects, 4 physically healthy and 6 brain damaged psychiatric patients formed the basis for the investigation of the properties of this clock. It was found that this perceptual clock is endogenous, self-sustaining and innate. Its free-running oscillations are independent of psychic state, emotions and affect. They are, however, dependent upon the subject's neural integrity and their phase may be influenced by an orientating reflex. Timing is precise but the preceding phase may influence a succeeding one. Despite the muscular activity of the task the clock is relatively independent of body temperature. It is also immune to most psychoactive drugs.


Interindividual differences in circadian patterns of catecholamine excretion, body temperature, performance, and subjective arousal. Interindividual differences in circadian rhythms of urinary catecholamine excretion, performance, self-ratings of arousal and oral temperature were studied in 80 subjects divided into three groups--morning-active, evening-active, and intermediate. Catecholamine excretion, body temperature, and self-ratings of arousal exhibited pronounced circadian variations. Morning-active subjects exceeded other groups in the 24 h level of adrenaline excretion but crest phases did not differ, occurring close to 13.00 h. No differences between groups were found for noradrenaline excretion. Crest phases occurred close to noon. Self-rated alertness exhibited a significantly earlier (14.12 h) crest phase for morning-active than for evening-active subjects (16.09 h). The performance did not differ between groups.


Catecholamine correlates of radar monitoring performance. Two experiments were concluded in which target detection performance was measured for male and female subjects engaged in a simulated radar monitoring task. In the first, 20 subjects performed for 2 h; in the second, 28 subjects for 1 h. Adrenaline excretion rate was significantly correlated with performance efficiency in both cases. Task duration and subject sex had no effect upon that correlation. The results of the two experiments were combined to show a general adrenaline/performance relationship.


Electrodermal conditioning to potentially phobic stimuli in male and female subjects. There are clear sex differences in incidence of phobias for small animals, and in questionnaire-measured fear for animals. The present study examined whether these sex differences were reflected also in electrodermal conditioning to potentially phobic stimuli. Separate groups of males and females were exposed to a conditioning session involving either potentially phobic, snakes and spiders, or fear-irrelevant, flowers and mushrooms, conditioned stimuli with electric shock as the unconditioned stimulus. A long interstimulus interval differential paradigm was used, allowing analysis of first- and second interval anticipatory responses, and third-interval omission responses. There were 8 habituation trials, 16 acquisition trials and 40 extinction trials. Half of the trials involved the reinforced cue, and the other half the unreinforced cue. There were clear conditioning effects, with superior acquisition and resistance to extinction to the phobic as compared to the fear-irrelevant stimuli. However, there were no differences between the two sexes. The results were interpreted in terms of the preparedness theory of phobias, and in terms of social learning factors.


Development of a short term assay for lymphoblastic transformation in man. Blastic stimulation of lymphocytes by antigens or mitogens is usually measured by 3H-TdR labelling of DNA synthesis in 4 to 7 days. However, it has been recently shown, first in mice and guinea pigs, then in men, that it is possible to detect an earlier event by measurement of protein synthesis with 3H-Leucine in a few hours. The work presented here is devoted to determine the optimal conditions of such a short term assay in man. The technique we propose is described. Preliminary clinical results in lymphomas patients, compared to "in vitro" skin tests, are encouraging.


Neonatal polycythemia in low birthweight infant. 74 low birthweight infants haematologic findings were studied during the first 24 hours and at the 8th day of life. By statistical methods derived from the multivariate analysis: stepwise multiple regression analysis and factor analysis, correlations were investigated between haemogram data (red cells count, haemoglobin and haematocrit levels and 18 others parameters. Most of the infants with polycythemia have toxaemic mothers and were born after a long gestation: they are affected by intra-uterine growth retardation, have a low glycaemia level but a normal calcaemia level, a large skull perimeter and a small thymus volume. But no correlation was observed between polycythemia and the following data: heart volume, neonatal icterus intensity, reticulocytes and erythroblasts counts.


Is there evidence for subclasses of chronic lymphocytic leukemia? A study using numerical classification techniques. Various clinical and biological parameters were analysed among 95 chronic lymphocytic leukemia patients by two numerical classification techniques in an attempt to identify natural subdivisions in the disease. For each patient, a profile made up of 47 dichotomous variables was constructed and the distance between profiles computed with equal weight for each variable. No specific cluster of patients was identified by these techniques. However limited availability of data for some laboratory tests, particularly concerning the immune system, meant that these results had to be excluded from analysis. The profiles of patients who died from leukemia during a mean follow up period of 4 years apparently do not generate a distinct group from the other profiles in spite of some clustering.


Ultrastructure, biological study and culture of C-cell carcinoma of the thyroid gland. The ultra-structural of a C-cell carcinoma of the thyroid confirms previous data: the cells are slender, with a large nucleus. The Golgi apparatus is well developed. Granules in striking abundance are seen in every part of the cytoplasm: their diameter is 85 - 250 mmu.


A long-term effect of adult thymectomy on cortisone sensitivity and alkaline RNAse activity of murine lymphoid cells. The alkaline RNAse content of lymphoid cells was measured in two year old mice that had been thymectomized at six to seven weeks of age and found to be three times higher in spleen and four times in lymph nodes than in sham thymectomized age-matched controls. There was, however, no difference in the content of alkaline RNAse in the liver between the two groups of mice. Following administration of cortisone, the RNAse level in the lymphoid organs was increased to a much greater extent in the control than in the thymectomized mice.


Uptake of aluminium ion by the liver.

Uptake of aluminium ion by the liver. The specific uptake by the rat liver of 28Al was shown. There was specific uptake of 28Al by liver cell nuclei and DNA. This uptake was blocked by prior treatment with stable aluminium. It is concluded that aluminium enters the liver cell by some specific mechanism and that the reaction of aluminium with DNA inside the hepatocyte nucleus could be a mechanism responsible for the development of aluminium-induced experimental porphyria in the rat.


Bilirubin and paranitrophenol glucuronyl transferase activities and ultrastructural aspect of the liver in patients with chronic hemolytic anemias. The hepatic ultrastructural aspect and the hepatic bilirubin (Bil-GT) and paranitrophenol (PNP-GT) glucuronyl transferase activities were studied in twenty subjects with a chronic hemolytic anemia (HA) and five subjects with an aplastic anemia. In chronic HA: the hepatic ultrastructural aspect showed mitochondrial abnormalities affecting structure (paracrystalline inclusions), size (giant mitochondria) and shape (irregular mitochondria); there was a decrease of Bil-GT activity in 80 percent of the patients. No connection could be shown between the HA etiology and the degree of the enzymatic activity decrease. The physiopathological mechanism of this enzymatic activity decrease is unknown; no deficiency of PNP-GT was observed; in chronic aplastic anemias there does not seem to be any modification, either of Bil-GT activity, or of hepatic PNP-GT activity. It is concluded that Bil-GT is often decreased in HA and that neither Bil-GT measurement nor the ultrastructural aspect of the liver distinguish chronic HA from Gilbert's syndrome.


Alteration in the uptake and storage of cardiac noradrenaline in spontaneously hypertensive rats. Uptake and storage of noradrenaline administered to Spontaneously hypertensive rate (SHR) (14) was significantly reduced compared with normotensive Wister rats (NR). Initial uptake of noradrenaline at 10 minutes was only 19 per cent less in SHR than in NR. However, 60 minutes following the administration of noradrenaline, its concentration in the heart of SHR was 60 per cent less than that in NR. Noradrenaline administered to NR disappeared from the heart in 10 hours, and that from SHR in 5 hours only.


Fetal intestinal disaccharidases in human amniotic fluid. Based on data from two human fetuses, a correlation exists between amniotic fluid and intestinal disaccharidase activities suggesting that the fetal intestine is the source of these amniotic fluid enzymes. Intestinal disaccharides are apparently released into the amniotic cavity by fetal defecation. As indicated by the drop of disaccharidase activities in amniotic fluid at 22 weeks gestation, defecation stops at this period of gestation.


Measurement of cell growth by a rapid U.V. spectrophotometric method. A spectrophotometric method for measuring mammalian cell growth in culture is described. The attached cells were solubilized in 2N sodium hydroxide and the absorption was measured at 225 nm. The technique is sensitive to about 3000 cells per Petri dish and a linear relationship was obtained between the number of cells and the absorption at this wave length. The method is rapid and the solubilized material can be recovered for liquid scintillation counting when needed.


Plasma prolactin response to L-dopa TRH and metaclopramide in thyrotoxicosis. The plasma PRL repsonse to synthetic TRH, L-dopa and metaclopramide was evaluated in patients with thyrotoxicosis. A PRL response similar to thtaclopramide,while no PRL response was observed after TRH. When the TRH-test was repeated after treatment, all patients showed a normal increment in plasma PRL. This data suggests that, in thyrotoxicosis, PRl response to TRH only is impaired, but PRL can be suppressed by L-dopa or released by metaclopramide notwithstanding the elevated values of thyroid hormone levels.


Bidimensional immunoelectrophoresis in three stages (semi micromethod). A three stage bidimensional immunoelectrophoresis enables a single protein to be identified in a complex rocket pattern. This identification is due to evidence of a complete reaction of identity between the antigen-antibody precipitation lines obtained in two gels.


Anti-adrenocortical activity of SKF 12185 and SKF 252 A. The synthesis of aldosterone and 18-hydroxycorticosterone from triated corticosterone by sheep adrenal homogenate have been studied in presence of increasing concentrations of SKF 12185, a product already known as an inhibitor of cortisol synthesis. An approximately 98% inhibition of the synthesis of both steroids occurred with only 2 x 10(-4) M of the drug. In contrast, incubation of a structurally closely related compound (SKF 252 A) under the same experimental conditions resulted in an only 52% inhibition with higher concentrations (2 x 10(-3) M). These findings could explain the marked aldosterone fall that occurs in SKF 12185 treated subjects and supports the therapeutic use of this product in treatment of hyperaldosteronism.


A possible physiological function of lysozyme. A possible physiological function of lysozyme might consist of an indirect effect arising in the course of its action on bacteria, as some of the bacterial reaction products behave as adjuvants or immunostimulating substances.


Elevated CEA levels in polyvinyl chloride workers. Plasma CEA titer was measured in 200 polyvinyl chloride workers. A positive test was obtained in 48.3% of this high risk population as compared to 9.2% for a normal healthy population.


Effect of 24 hour fast in obese children. Effects of a 24 hour fast were studied in 21 obese children aged 7 to 14 and in 8 controls. Mean blood glucose (BG) during fast dropped more in controls (0.88 to 0.54 g/l) than in obese (0.90 to 0.63 g/l) Plasma cortisol changes were similar in the 2 groups, FFA increased (p less than 0.01) in the 2 groups, but the 24 hour mean level was higher in controls (4.0 mEq/l) than in obese (2.06 mEq/l). At the end of the fast, a ketonuria was present in all obese children except 2. Serum alanine dropped similarly in obese (28 to 24 muM p. cent ml) and in controls (30 to 22 muM p. cent ml). All obese exhibited at the end of the fast a significant rise (p less than 0.01) of branched chain aminoacids, not observed in controls. Responses to glucagon (0.03 mg/kg I.M.) were studied before and after fast. At time 0, BG response was higher and more prolonged in obese in spite of hyperinsulinism. At time 24 hours, BG raised from 0.50 to 0.74 g/1 and insulin from 8 to 35 muU/ml in controls, while in obese BG raised from 0.63 to 1.06 g/l and insulin from 25 to 88 muU/ml. Concomitant hyperinsulinsim and biological criteria of hypoinsulinism demonstrated in obese children the peripheral resistance to insulin. The contrast between a normal degree of protein gluconeogenesis and a reduced rate of fat mobilization during fast may be a major biological feature of obesity in childhood.


Study on the composition of casein from individual human milks. The study of the chemical composition and of the electrophoretograms of 21 different human whole casein samples confirms the presence of an important heterogeneity between the different samples. A very considerable variability of phosphorus and sugars contents and a correlation between the different sugars components of the casein have been demonstrated statistically. The study of 3 milk samples from the same woman showed a decrease of the amino-sugars of the casein after the birth.


Miniature chromatography of haemoglobin A2 and heterozygous beta thalassemia. 172 beta thalassemias are detected by applying a new method of haemoglobin A2 determination: Miniature chromotography on DEAE cellulose. This procedure is simple, fast, accurate and there is no significant difference with macro-chromatography. It is proposed as standard method for determination of hemolgobin A2. This study was made on 400 patients suspected of beta thalassemia, the majority of which presented with a considerable microcythemia.


Relationship of blood and saliva lactate and pyruvate concentrations. Lactate and pyruvate concentrations of blood and saliva were investigated in nine healthy young men before and after maximal exercise. Lactate and pyruvate concentrations in saliva and blood were found to be unrealted. Interpretation favored salivary gland anaerobic metabolism as the origin of salivary lactate, but the large difference of 80 mg per 100 ml between blood and saliva found after 12 min of recovery may represent a threshold value and leaves unanswered the question on the origin of lactate in saliva. Multiple sampling following the recovery period for a longer time may change the present interpretation.


Artefacts due to sodium dodecylsulfate polyacrylamide gel electrophoresis in the study of human erythrocyte membrane calcium binding protein. Sodium dodecylsulfate polyacrylamide gel electrophoresis is not usable for identification of calcium binding protein from human erythrocyte membrane; Ca2+ binds to anionic sites of SDS and with 45Ca radioactivity bands appear in gels in absence of proteins.


Rat leukaemia cell line grown on a chemically defined medium. Leukaemia cells of C12/0 line, derived from Gross virus-induced W/Fu rat thymoma, have been successfully maintained in a completely synthetic, serum-free RPMI 1640 medium. The cells propagated as good as in serum-supplemented medium, though they could not grow from a low cell density. Cells adhered to the substratum and did not survive. This problem was overcome by use of silicone-coated substratum, or of microexudate from C12/0 cells adsorbed on the substratum. The cells underwent degeneration soon after they reached their maximum density. C12/0 cells released macromolecules might possibly have both growth-promoting and toxic effect on the cell itself.


Primary carcinoma of the colon and hyperlipemia: a paraneoplastic syndrome. The human counterpart of the association of hyperlipidemia with cancer is rare, as compared with the relative frequency of the syndrome in experimental animals. A case is presented of adenocarcinoma of the colon with a non casual relationship between the presence and the progression of the tumor and hyperlipemia. Surgical resection and antiblastic chemotherapy moreover seemed to influence the metabolic anomaly.


Oestrogen receptors in male breast cancer. Eleven samples from 7 primary and 4 metastatic male breast cancers were analysed for cytoplasmic oestrogen receptors. Two samples of metastatic cancers were obtained from a same patient 6 months apart. Receptors were assayed by measuring the binding affinity of cytosol fractions for 3H-oestradiol- 17beta. Saturable binding sites were obtained in 6 primary and 1 metastatic cancers. The dissociation constant of the binding reactions were within the range reported for the specific oestrogen receptors from female breast cancers. When present, receptor concentrations varied from 59 to 532 fmoles/mg tissue protein. Competition studies of the binding of 3H-oestradiol- 17beta by excess amounts of unlabelled steroid hormones or antihormones revealed that the receptors were specific for oestrogens and antioestrogens. By all these criteria, receptors from male mammary cancer were identical to those from female breast cancer. Chromosome analysis was performed in 6 patients. All had normal 46 XY male pattern indicating that no conspicuous genetic abnormalities such as those of the Klinefelter syndrome seemed responsible for the presence of the receptors.


Study of two cases of bisalbuminemia by electrofocusing. Isoelectric focusing was used to compare normal seric albumin and seric albumin from two subjects with bisalbuminemia. An additional new albumin fraction was found in each of these two anormal sera. The isoelectric point of these variants was respectively 5.65 and 5.84. These results suggest that isoelectric focusing may be used to identify and classify the different variants of albumin.


Immunological study of hydatidosis. I. Evaluation of the test of immuno-electrodiffusion in the humoral study of human hydatidosis. The importance of immunological arguments in the discussion of hydatidosis, the permanent increase of requirements, the risk of cross reactions, the frequency of sero-epidemiological surveys imply the utilisation of specific and rapid micro-methods. The technique of immunoelectrodiffusion on a cellulose acetate membrane answers partly to those criteria; we have used it since 1971 in the diagnosis of parasitic infections and mainly to that of hydatidosis. Two hundred cases (400 samples) of the latter were studied longitudinally and reported. This micro-method is very specific (co-immunoelectrodiffusion), equal to that of immunoelectrophoresis. Its rapidity (3 to 4 hours) is close to that of indirect immunofluorescence. The revelation of suggestive diagrams of fissuration underlines the relative emergency of the definitive interpretation of an immunological examination in the diagnosis of hydatidosis.


Duodenal calcium-binding protein (CaBP) and phosphorus deprivation in growing rats. The present study has been undertaken to elicit the effects of a low-phosphorous diet on intestinal calcium absorption. Measurements of duodenal calcium-binding protein (CaBP), mucosal calcium, fractional calcium absorption and duodenal calcium transport capacity has been performed in growing rats fed for three weeks a low-phosphorous diet adequate in vitamin-D3. In animals on low-phosphorous diet, the mucosal concentration of CaBP measured by a quantitative competitive binding assay has increased in parallel with the mucosal calcium concentration, the fractional absorption and the calcium transport capacity. Analytical gel electrophoresis of calcium-binding protein in animals fed low-phosphorous diet has shown the presence of a protein band which would involve more than the effect of the stimulated biosynthesis of 1alpha,25-DHCC.


Chemical ionization mass spectrometry of polyunsaturated fatty acids of human serum. Polyunsaturated fatty acids of human serum were analysed by combination gas-liquid chromatography and chemical ionization mass spectrometry, using isobutane as the reagent gas. The method described yielded the molecular ion of long chain unsaturated fatty acids as the base peak; these were weak or unobservable in the corresponding electron impact mass spectra. In the chemical ionization mass spectrometry, the derivatization with trimethylsilyl ether was demonstrated to be useful for the location of the double bonds of unsaturated fatty acids. Applied toteh serum polyunsaturated fatty acids, these methods have given identification of C18:4, C20:4, C20:5, C22:4, C22:5omega-6, C22:5omega-3, and C22:6.


The structure of narasin and a related ionophore. Using electron impact and field desorption mass spectrometry, structures are proposed for narasin and a related ionophore, A28086B, derived from fermentation. The ionic fragmentation of these compounds and their derivatives is discussed and compared with that of salinomycin, whose structure has been determined by X-ray diffraction.


Electron impact mass spectrometry of some ketosugar derivatives. The electron impact fragmentation of 1,2-O-isopropylidene-alpha-D-glycero-tetros-3-ulofuranose has been studied with the help of deuterium and 18O labelling. The effect of substituting a hydrogen atom of C-4 for a CH3 radical and that of replacing the carbonyl function with a cyanomethylenic substituent is also reported.


Determined of bufuralol and its metabolites in plasma by mass fragmentography and by gas chromatography with electron capture detection. A sensitive method for the simulataneous determination of bufuralol and its pharmacologically active metabolites in huamn plasma is described. The O-timethylsilyl,N-trifluoroacetyl derivatives are assayed by mass fragmentography. Sensitivity is 1 ng 76(-1) plasma for bufaralol and about 250 pg ml(-1) for the metabolites. An alternative procedure which uses gas chromatography with electron capture detection is also described. The sensitivity of this is about 10 ng ml(-1) plasma for all drug-related components.


The direct determination of oestrogen and progesterone in human ovarian tissue by quantitative high resolution mass spectrometry. The concentration of oestrone, oestradiol and progesterone in ovarian tissue have been determined directly by quantitative high resolution mass fragmentography without the necessity of extraction or purification. The changes in the levels of these steroids in the ovary throughout a 28 day period correlate well with those measured in plasma and also with those of the corresponding metabolites excreted in urine. Some benign ovarian tumours (cysts) have been shown to contain abnormally large amounts of progesterone.


Aromatic hydroxylation of ellipticine in rats: lack of an NIH shift. Metabolites of the antitumor agent, elipticine (NSC) 71795), are mainly secreted in rat bile as two conjugates of hydroxylated elipticine. The possible involvement of an arene oxide intermediate prior to hydroxylation and conjugation has been studied using 7,9-dideutero-ellipticine and by determining the conservation of deuterium in the phenolic products. The deuterated drug was synthesized by acid catalyzed exchange and was characterized by nuclear magnetic resonance and high and low resolution mass spectrometry. Mass spectra of two conjugated metabolites from rat bile showed a mass ion at m/e 263 (singly deuterated hydroxylated elipticine). There was no evidence for conservation of deuterium at the position of hydroxylation (m/e 264). These results indicate that the major pathway of metabolism for this cancer chemotherapeutic drug, prior to conjugation and excretion in rat bile, proceeds by aromatic hydroxylation apparently without involving an arene oxide/NIH shift mechanism.


A model for ionization mechanisms in field desorption mass spectrometry. The apparent discrepancy between theory and experimental practices in field desorption mass spectrometry has encouraged a study attempting to focus theoretical concepts more clearly into forms recognizable to the chemical investigator. Preliminary results indicate that best anode temperature for many compounds is independent of the applied voltage throughout the range available on the Varian MAT CH-5 double focusing mass spectrometer with the electron impact-field ionization-field desorption combined source. Studies of mass transfer indicate that desorption of the sample at the best anode temperature occurs in the absence of the applied field and at a rate which is indistinguishable from the rate of desorption in the presence of the field. These results and other observations, have led us to postulate a new mechanism of ion formation, involving chemical attachment reactions in a thin fluidized system. Clearly, the role of the field in this technique must be the subject of continued study.


Monocyte-mediated antibody-dependent cellular cytotoxicity: a clinical test of monocyte function. The lack of a simple, rapid, and quantitative test of the functional activity of the monocyte has hampered studies of the contribution of this cell type to host defense and human disease. This report describes an assay of antibody-dependent cellular cytotoxicity, which depends exclusively upon the monocyte as the effector cell and therefore provides a convenient test of monocyte function. In this system, mononuclear leukocytes (MNL) obtained by Ficoll-Hypaque separation of whole blood are cytotoxic for 51Cr-labeled human erythrocyte targets coated with anti-blood group antibody. Removal of phagocytic monocytes from the MNL by iron ingestion, followed by exposure to a magnetic field, completely abolishes all cytotoxic activity from the remaining MNL population. Similarly, in severely mono-cytopenic patients with aplastic anemia, cytotoxic effector activity is absent. In normals and less severely monocytopenic aplastic anemia patients, cytotoxicity correlates significantly (p less than 0.001) with monocyte number. Application of this monocyte-mediated antibody-dependent cellular cytotoxicity assay to the study of patients with the Wiskott-Aldrich syndrome has revealed defective monocyte cytotoxic activity in spite of normal monocyte numbers, suggesting that this test may be useful for the assessment of monocyte function in a variety of clinical situations.


Steroids and hematopoiesis. III. The response of granulocytic and erythroid colony-forming cells to steroids of different classes. Selected androgenic and nonandrogenic steroids enhance in vitro granulocytic and erythroid colony formation by mouse marrow cells, but do so by influencing either different target cells or cells in different states of cell cycle. Etiocholanolone, a naturally occurring nonandrogenic testosterone metabolite, permits cells not in active cycle to respond to colony-stimulating factor or erythropoietin. Fluoxymesterone, a synthetic androgen, appears to enhance colony growth by increasing the responsiveness of target cells to tropic stimuli. The majority of cells responding to this androgen are in active DNA synthesis. Direct comparison, however, of etiocholanolone-dependent erythroid or granulocytic colony-forming cells demonstrates nonidentity of the target cells. Thus colony-forming units responding to different classes of steroids are in different states of cell cycle and are physically separable. The enhancement of the in vitro response of colony-forming cells to regulating hormones by steroids such as etiocholanolane suggests a mechanism by which such agents may be therapeutically effective in certain cases of marrow failure in man.


The effect of chelating agents on iron mobilization in Chang cell cultures. The investigation of chelating agents with potential therapeutic value in patients with transfusional iron overload has been facilitated by the use of Chang cell cultures. These cells have been incubated with [59Fe]transferrin for 22 hr, following which most of the intracellular radioiron is found in the cytosol, distributed between a ferritin and a nonferritin form. Iron release from the cells depends on transferrin saturation in the medium, but when transferrin is 100% saturated, which normally does not allow iron release, desferrioxamine, 2,3-dihydroxybenzoic acid, rhodotorulic acid, cholythydroxamic acid, and tropolone all promote the mobilization of ferritin iron and its release from cells. They are effective to an approximately equal degree. The incubation of [59Fe]transferrin with tropolone in vitro at a molar ratio of 1:500 results in the transfer of most of the labeled iron to the chelator, reflecting the exceptionally high binding constant of this compound. How far these phenomena relate to therapeutic potentially remains to be seen.


Fitzgerald factor (high molecular weight kininogen) clotting activity in human plasma in health and disease in various animal plasmas. Fitzgerald factor (high molecular weight kininogen) is an agent in normal human plasma that corrects the impaired in vitro surface-mediated plasma reactions of blood coagulation, fibrinolysis, and kinin generation observed in Fitzgerald trait plasma. To assess the possible pathophysiologic role of Fitzgerald factor, its titer was measured by a functional clot-promoting assay. Mean +/- SD in 42 normal adults was 0.99+/-0.25 units/ml, one unit being the activity in 1 ml of normal pooled plasma. No difference in titer was noted between normal men and women, during pregnancy, or after physical exercise. Fitzgerald factor activity was significantly reduced in the plasmas of eight patients with advanced hepatic cirrhosis (0.40+/-0.09 units/ml) and of ten patients with disseminated intravascular coagulation (0.60+/-0.30 units/ml), but was normal in plasmas of patients with other congenital clotting factor deficiencies, nephrotic syndrome, rheumatoid arthritis, systemic lupus erythematosus, or sarcoidosis, or under treatment with warfarin. The plasmas of 21 mammalian species tested appeared to contain Fitzgerald factor activity, but those of two avian, two repitilian, and one amphibian species did not correct the coagulant defect in Fitzgerald trait plasmas.


Evidence suggesting the regulation of a coagulation factor levels in rabbits by a transferable plasma agent. New Zealand white rabbits were given 30 ml of goat serum intravenously. This procedure resulted in an immediate decrease in platelet count, fibrinogen, and levels of coagulation factors II, V, VII, and X, due to consumption coagulopathy. These factors returned toward baseline levels approximately 12 hr after the injection. Plasma from rabbits who had received goat serum 48 hr previously (donor rabbits) was injected into recipient rabbits. This procedure resulted in a slight rise in the level of coagulation factor II (range, 20%-30%) and a significant rise in factors V (35%-75%), VII (35%-235%), and X (35%-75%) in the recipients. When plasma from control donor rabbits who had not received goat serum was injected into recipients, there was no change in these coagulation factors. It is postulated that the reduction in coagulation factor levels in donor rabbits induces a "coagulopoietin" for each factor or one "coagulopoietin" for all factors which stimulates increased synthesis and/or release of these factors in recipient rabbits.


[Longtime therapy of congenital factor XIII deficiency using factor XIII concentrate]. Factor XIII was determined by enzymatic and immunochemical methods in 3 patients with congenital factor XIII deficiency. Factor XIII activity measured by trans-glutaminase assay was below 1% of normal value in each of these cases. Immunelectrophoresis determination revealed the absence of the functionally active subunit A, whereas subunit S was only slightly diminished (30 to 50% of the normal value). Substitution with factor XIII concentrate caused a parallel increase of factor XIII activity and subunit A concentration. No uptake of factor XIII activity or of subunit. A by platelets could be demonstrated. Despite discontinuous substitution over a period of six years no antibody against factor XIII activity could be demonstrated in one patient with congenital factor XIII deficiency.


Neutrophil marrow release. A system analysis. Neutrophil marrow egress is governed by several processes. The most important are cell maturation, functional behavior of marrow sinusoids and humoral or neuro-vascular factors. Neutrophil release cannot be observed directly but is reflected in the size, cellular composition and kinetics of the nonproliferating pool of granulocytopoiesis in bone marrow and of blood neutrophil pool. These experimentally determined parameters were used as the basis of a mathematical model study. The model describes two catenated compartments, the nonproliferating pool of granulocytopoiesis in marrow and the total blood granulocyte pool. Cell transit from one pool to the other was assumed to be age-dependent. It was expressed by a positive sloping sigmoidal function that defines the egress potential fo the cells that increases with cell maturation. During maturation granulocytopoietic cells develop intense motility which determines the morphology of the cells on smears. Relationship between cell motility and its morphology was defined by functions determining the age-dependent probabilities of cell fixation as metamyelocytes, band- and segmented forms, respectively. The parameters of this model could be so adjusted that all experimental data were matched within experimental errors. Thus, qualitative and quantitative information on neutrophil marrow egress was obtained for normal and pathological states of granulocytopoiesis.


Tuberculosis control in the Americas: current approaches. Tuberculosi remains a serious public health problem in the Americas, and it has not declined as rapidly or as much as experts projected it would in the 1940's. Scientific advances in control of the disease over the last three decades have produced effective chemotherapeutic agents, established the immunizing capacity of BCG vaccine, and demonstrated the superior value of bacteriologic diagnosis in symptomatic individuals over mass community x-ray surveys, which are both inefficient and costly. They have also shown that most cases can be treated on an ambulatory basis, obviating the need for the lengthy hospital stays which have heretofore weighed so heavily on budgets. By standardization of control methods, both for diagnosis and for chemotherapy, these tasks can be taken on by polyvalent staff in the general health services, whose wide coverage places them in a position to reach a much larger segment of the population than that attended by the traditional vertical system. To a greater or lesser degree, all the countries in the Americas are beginning to orient their strategies in this direction, and some of them already have considerable progress to report.


Measles vaccination.

Measles vaccination. Prevention of measles poses different problems in developed and developing countries. In developed countries the risk of late complications following the use of live vaccine should be further evaluated; additional attempst might also be made to create new types of inactivated vaccine. In developing countries the introduction of live measles vaccine should be given high priority. The main problems in developing areas appear to be ones relating to vaccine stability, distribution, and cost.


Rubella vaccination.

Rubella vaccination. Various attenuated live rubella vaccines now in use are considered generally safe and immunogenic, but to date the longest that vaccinal immunity has been shown to endure is seven years. Subclinical reinfection is not uncommon among vaccines, but its effect on pregnancy and fetal development is not yet fully known. At present two mass immunization strategies are used: vaccination of all children under 12 years of age or protection only of females 10-14 years of age. In either case susceptible women of childbearing age who have no detectable rubella HI antibody are immunized as well. Such women may receive the vaccine during the postpartum period or at any other time when they are not pregnant, but strict precautions must always be taken to make sure they do not conceive for at least two months thereafter.


[Experimental terato-carcinoma: behavior of embryoid bodies inoculated into the abdominal cavity of syngeneic mice]. A suspension of CE "44" embryoïd-bodies obtained from a transplanted OTT 60-50 subline was injected into the abdominal cavity of syngeneic L 129 mice. Terato-carcinoma and ascitic fluid were produced the latter containing embryoïds. In situ examinations were made by serial transverse sectioning of the entire abdomen. The reconstruction of the sequence of events occuring after inoculation was achieved by killing the animals at different time after inoculation. This analysis showed that embryoïd-bodies appear quite similar to blastulas and that they are attracted to evolving granulomas in the peritoneum. Blastulas attach through their outer wall which then breaks to permit the inner-cell masses to penetrate through the hole and contribute towards the subsequent development of terato-carcinomas.


[Value of the cytologic diagnosis of adenopathies by aspiration biopsy. Cytohistologic comparison of 1756 cases]. The value of the cytologic diagnosis by aspiration needle biopsy is analysed in a series of 1756 adenopathies, verified by histological examination (1311 malignant nodes and 445 inflammatory or normal lymph nodes). The result is that a large number of lymph node tumours, either primary or secondary, are identified by the cytologic examination (from 77% to 91% of the cases according to the explored zone). Furthermore the virtually non existent percentage of false positive results (3% of the cases) proves the great accuracy of this method in the diagnosis of malignancy. In contrast, the diagnosis of benign nature must never be stated without biopsy (2% to 4% of the cases are erroneous by negative according to the explored zone). Indeed, even if benign cells are obtained, nothing allows to eliminate a malignant tumour or a limited inflammatory zone, not reached by the needle.


Health education in rural Subinam: use of videotape in a national campaign against schistosomiasis. A videotaped program on schistosomiasis, coordinated with a wide range of other campaign activities, was shown at schools, clinics, and other gathering places in the rural Surinam district of Saramacca. The usual procedure was to have two showings, the first for teachers and the second for pupils in grades four through six. The children were then asked to answer simple questions about the disease in writing. Booklets were also distributed to teachers and children, and in a number of instances follow-up activities were carried out such as classroom exercises and in one case an art contest. By and large the tape was a success. Viewer attention was best when the audience was comfortably seated and undistracted. It was found that the viewing by itself was not enough to ensure basic understanding of the schistosomiasis problem. Writing about the event afterwards helped for it to be taken seriously, reinforced the information conveyed, and allowed children to relate the ideas presented to their own experiences. It also gave the campaign workers a basis on which to estimate the degree of success achieved. At the same time, follow-up projects such as the art contest enlisted the active participation of children, community members, and educators and added greatly to the overall educational effect.


Combined modality approach in small cell carcinoma of the bronchus. A pilot study. Our first experiences with polychemotherapy (COMB-regimen) followed by radiotherapy for patients with small cell carcinomas are encouraging. 7 patients out of 18 showed a complete remission. The treatment was well tolerated by all patients.


Community psychiatry and the Pan American Health Organization: the Jamaican experience. The development of a modern psychiatric service in Jamaica and the role played by PAHO consultants in this development is described. Some fundamental points are raised: -It is essential that a director of mental health services and a senior psychiatric nurse be given senior positions in the Ministry of Health so that the psychiatric services receive due attention and are integrated closely with the other health services. -The local training of psychiatric professionals of all categories is essential for such a service and should be given priority by the Government. -The intelligent use of a large number of briefly trained individuals is the only way to provide psychiatric coverage in a developing country. -PAHO consultants assigned to work in a developing country over an extended period can play a useful role in the introduction of contemporary psychiatric services.


[Clinical peculiarities of nasopharyngeal cancers]. Nasopharyngeal carcinomas are very different from other malignancies of the upper digestive and respiratory tracts. Very uncommon in France, mostly undifferenciated, they occur in all decades, even in children or young adults, with a sex-ratio of 3/1, and without connection with alcohol and tobacco. Their insidious onset leads to the first manifestation as cervical metastasis, or evidence of intracranial involvment. The 5 years Survival rate is 30 p. 100: failures are local or distant metastases, principally in the skeleton. Destruction of the base of the skull and supraclavicular nodes yield poor results. Trials on systemic chemotherapy are in progress.


A simple technique to treat drooling: lip resection and muscle flaps. a simple technique to correct the lax oral sphincter has been developed. A wedge resection of the bulky, atonic lower lip is performed leaving a pedicle flap of orbicularis oris muscle on either side of the resection. The muscle flaps are tunnelled submucosally toward the opposit commissures and anchored. This gives excellent stability to the lower lip and prevents drooling.


A simple diagnostic sign in the superior mesenteric artery syndrome in a burned patient. In the superior mesenteric artery syndrome which may complicate extensive burns, a systolic murmur may be heard in the epigastrium when the patient is supine but not when he is prone. This sign combined with straight abdominal x-rays will clinch the diagnosis and thereby avoid an unnecessary barium meal or laparotomy.


Lymphaticovenous anastomosis by microvascular technique. After developing some basic techniques of lymphaticovenous anastomoses in 60 dogs, a further study of these anastomoses in the greyhound was performed with a patency rate of 74 per cent a 1 week and 66 per cent between 6 and 12 weeks. Points of technique and causes of failure are discussed. The use of this method in the treatment of early secondary lymphoedema is mentioned.


A review of 102 cases of hypospadias treated by the Van der Meulen procedure. One hundred and two cases of hypospadias without chordee which were corrected by van der Meulen's operation were reviewed. There were no fistulas and only 10 complications. The functional result was good in 90 per cent.


Inhibitory innervation of cat sphincter of Oddi. 1 Electrical stimulation with trains of 0.1-0.2 ms pulses of the cat isolated sphincter of Oddi inhibited the spontaneous contractile activity and lowered base-line tension considerably. A contraction usually followed the period of stimulation. 2 These inhibitory effects were prevented by tetrodotoxin 0.1-0.5 mug/ml but were not reduced by hexamethonilm, morphine, or blockade of alpha- or beta-adrenoreceptors of cholinoceptors with phenoxy-benzamine propranolol or atropine, respectively. 3 Adenosine-5'-triphosphate (ATP) and adenosine-5'-diphosphate (ADP) inhibited the spontaneous sphincter activity and caused relaxation thus mimicking the effects of the C-terminal octapeptide of cholecystokinin (C8-CCK), isoprenaline and prostaglandin E1 and E2. 4 ATP alone (greater than 100 mug/ml) or ATP (greater than 10 mug/ml) plus dipyridamole (1 mug/ml), relaxed the sphincter to the same degrees as did the field stimulation. 5 In sphincter maximally contracted by acetylcholine, the effect of stimulation was more marked than that recorded in uncontracted preparations. 6 The present findings suggest that the sphincter of Oddi receives inhibitory nerves that are neither cholinergic nor adrenergic.