Freezing temperature of finger skin.

Freezing temperature of finger skin. In 45 subjects, 154 frostnips of the finger were induced by cooling in air at -15 degrees C with various wind speeds. The mean supercooled skin temperature at which frostnip appeared was -9.4 degrees C. The mean skin temperature rise due to heat of fusion at ice crystallization was 5.3 degrees C. The skin temperature rose to what was termed the apparent freezing point. The relation of this point to the supercooled skin temperature was analyzed for the three wind speeds used. An apparent freezing point for a condition of no supercooling was calculated, estimating the highest temperature at which skin freezes at a given wind speed. The validity of the obtained differences in apparent freezing point was tested by an analysis of covariance. Although not statistically significant, the data suggest that the apparent freezing point with no supercooling decreases with increasing wind velocity. The highest calculated apparent freezing point at -15 degrees C and 6.8 m/s was 1.2 degrees C lower than the true freezing point for skin previously determined in brine, which is a statistically significant difference.


Drug modification of hypothermia induced by CNS glucopenia in the mouse. 2-Deoxy-D-glucose (2-DG), insulin, or norepinephrine (NE), when injected into the cerebral ventricles of conscious mice, produce decreased rates of O2 consumption and hypothermia. These changes are accompanied by hyperglycemia with 2-DG, hypoglycemia with insulin, and normoglycemia with NE. Desipramine blocks the reduction in body temperature and O2 consumption produced by each of these agents, but does not modify significantly their effects on plasma glucose. The latter suggests that the thermal and oxidative responses to central glucopenia can be dissociated from concurrent alterations in circulating glucose. Propranolol enhances the hypothermic response produced by administered 2-DG, insulin, or NE. Phentolamine, however, antagonizes the hypothermia only with NE, indicating the 2-DG and insulin are probably not acting through the release of endogenous NE.


Effects of arterial versus venous sampling on analysis of glucose kinetics in man. A compartmental model is presented to account for transient and steady-state changes in blood glucose concentration which result from transit through the forearm and hand in man. This model permits the inter-conversion of arterial and venous data and the derivation of arterial equivalent total body glucose models from venous data. Data were obtained from subjects in the basal state following a pulse injection of [1-14C]glucose tracer. An artery, an antecubital vein, and a dorsal vein of a heated hand (68 degrees C environment) were sampled. Blood transit time is shorter 0.3 vs. 1.0 min) and irreversible glucose loss is reduced (1.9 vs. 2.9%) in the heated hand preparation when compared to the antecubital vein preparation. Because of the smaller correction required and the smaller variation among individuals when heated hand rather than antecubital vein data are obtained, we suggest that for analysis of whole-body kinetics such data should be used along with the compartmental model correction when arterial data cannot be obtained.


Cerebellar pressor response in the dog.

Cerebellar pressor response in the dog. A fastigial pressor response has been elicited in the anesthetized mongrel dog. Stimulation within the rostral portions of this nucleus results in mean arterial pressure rises up to 150 mmHg above control. A proportional tachycardia is simultaneously evoked which may rapidly attain heart rates of 190 beats/min above control levels. Peak tachycardias immediately suside and often the heart rate declines below control values during stimulation while arterial pressure remains elevated. When either the carotid sinuses were isolated by ligation or a bilateral vagotomy was performed, the fastigial tachycardia was sustained. Thresholds for the response are near 0.05 mA and the stimulus intensity curve is linear to 0.4 mA whereupon the pressure rise and tachycardia begin to approach a plateau value. The response could still be attained when submaximal doses of alpha-chloralose anesthesia or high levels of barbiturates (30-40 mg/kg) were given. Both portions of the response result from widespread sympathetic activation; however, buffering of the response through the baroreceptor reflexes is only demonstrated in the cardiac segment of the response.


The kinetics of rabbit muscle pyruvate kinase. Initial-velocity, substrate- and product-inhibition and isotopic-exchange studies of the reverse reaction. 1. An assay, based on the transfer of label from [gamma-32P]ATP to [32P]phosphoenolpyruvate, suitable for a steady-state kinetic analysis of pyruvate kinase in the reverse direction (i.e. phosphoenolpyruvate synthesis), is described. 2. This assay was used in a kinetic investigation of the rabbit muscle enzyme including initial-rate and product-inhibition experiments, at a pH of 7.4 and constant concentrations of total K+ and free Mg2+. 3. These studies indicate that there is a random release of ADP and phosphoenolpyruvate from the enzyme and that there is a competitive substrate inhibition by ATP. Some of the results were suggestive that the rapid-equilibrium assumption, generally used for this enzyme was not valid. 4. Techniques were developed to measure the rate of isotopic exchange between all the substrate-product pairs. 5. By using these techniques the rates of isotopic exchange at chemical equilibrium were measured. The results indicate that this enzyme does not catalyse a truly rapid-equilibrium random mechanism, although in the forward reaction all initial-rate data obtained to date are consistent with this assumption.


Effect of vagal tone on airway diameters and on lung volume in anesthetized dogs. In 18 open-chest dogs we obtained pressure-diameter (P-D) curves from tantalum bronchograms and pressure-volume (P-V) curves by plethysmography. After vagotomy most of the decrease in diameter with decreasing transpulmonary pressure (Ptp) occured below 10 cmH2O and there was no P-D hysteresis. Smaller airways narrowed more with decreasing Ptp than larger ones. Bronchodilators did not increase diameters after vagotomy (P less than 0.2). With vagi intact, diameters were smaller at all Ptp (P less than 0.01) and exhibited hysteresis, but the lung P-V curve was unchanged. Vagal stimulation narrowed airways further at all Ptp and hysteresis was marked. Smaller airways narrowed more with vagal stimulation than larger ones. Vagal stimulation did not change the deflation limb of the P-V curve but decreased inflation volumes slightly at all Ptp (P less than 0.01). We conclude that in vivo tone is vagal and that it affects the physical properties of airways, but not of lungs, making the airways remarkably independent from lung parenchyma.


Endotracheal tube as a factor in measurement of respiratory mechanics. The measurement of respiratory mechanics is frequently performed on intubated patients. This study shows the effect which the turbulent and inertial characteristics of an endotracheal tube have on the accuracy of the measurement. Two techniques are examined utilizing a mechanical model: the least-squares and the phasor technique. It is concluded that both techniques will give erroneous results if the characteristics of the tube are not accounted for. The error is markedly reduced if the turbulent and inertial terms are accounted for in the analysis.


Pyrenebutyric acid as an optical oxygen probe in the intact cerebral cortex. The application of the fluorochrome pyrenebutyric acid (PBA) as an in vivo quantitative probe for intracellular oxygen tension (iPO2) is reported. PBA is nontoxic, noninvasive, nonconsumptive, and allows optical monitoring of iPO2 on line. Cats were anesthetized with ether and lesioned midcollicularly to produce cerveau isolé preparations with optical monitoring via a compensated microfluorometer. The "normal" iPO2 was determined to be 42.8 /- 8.6 (SD) mmHg. Decrease in cerebral perfusion results in observing a decreased iPO2. This method of monitoring iPO2 was found to overcome drawbacks inherent in the polarographic technique and to posess additional advantages.


Measurement of tissue oxygen with a fluorescent probe. Evidence is presented that pyrenebutyric acid is nontoxic at the concentrations required for the measurement of fluorescence quenching by physiological concentrations of oxygen. It appears that this molecule can be used to measure tissue oxygen since it does not interfere with oxygen consumption. Equally, tissue components do not interfere with the quenching of fluorescence to such an extent as to invalidate the method. The spatial resolution of the technique is limited by some tissue geometrical factors which do not, however, appear to be great enough to prevent its use to measure intercapillary oxygen gradients.


Blood-perfused working isolated rat heart. We describe a method for perfusion of a working isolated rat heart with washed erythrocytes suspended in a Krebs-Henseleit bicarbonate buffer containing bovine albumin (fraction V). With washed pig red cells, as hematocrit was varied between 0 and 40%, coronary flow (CF), aortic flow (AF), external work (W), and myocardial oxygen consumption (MVO2) were measured. Hemodynamic data at a hematocrit of 30% (CF = 5.4 +/- 0.7 ml/min per g, AF = 75 +/- 8 ml/min per g) were identical with those reported for the intact animal. Coronary sinus PO2 was highest with a red cell-free perfusate suggesting that coronary flow is partially shunted. Human red cells obtained from banked blood, were tried also with success. With careful filtration, the preparation is stable for 2 h and well suited for study of the dynamics of myocardial oxygen delivery.


Methylated purines in the deoxyribonucleic acid of various Syrian-golden-hamster tissues after administration of a hepatocarcinogenic dose of dimethylnitrosamine. 1. DNA was extracted from livers, kidneys and lungs of Syrian golden hamsters at various times (up to 96h) after injection of a hepatocarcinogenic dose of [14C]dimethylnitrosamine. Purine bases were released from the DNA by mild acid hydrolysis and separated by Sephadex G-10 chromatography. 2. At 7h after dimethylnitrosamine administration liver DNA was alkylated to the greatest extent, followed by that of lung and kidney, the values for which were 8 and 3% respectively of those for liver. 3. The O6-methylguanine/7-methylguanine ratios were initially the same in all three organs and in the liver DNA of rats under similar conditions of dose. 4. O6-Methylguanine was the most persistent alkylated purine in all three hamster tissues. There was evidence for excision of 7-methyl-guanine, the highest activity for this being present in the liver. 5. Detectable amounts of the minor products 3-methyladenine, 1-methyladenine, 3-methylguanine and 7-methyladenine were present in most hamster tissues, and their individual rates of loss from liver DNA were determined. 6. Ring-labelling of the normal purines in DNA was highest in the liver, followed closely by the lung (80% of that in liver) whereas the kidney had very low incorporation (3% of that in liver). 7. The results are discussed with respect to the hepatotoxicity of dimethylnitrosamine, the miscoding potential of the various alkylation products and the induction of liver tumours in hamsters.


Rabbit muscle myogen. Interactions with phosphate as the source of non-enantiography in moving-boundary electrophoresis. Rabbit muscle myogen has been subjected to moving-boundary electrophoresis and velocity sedimentation in 0.0187 M-potassium phosphate buffer, pH7.7, I = 0.05. The ascending and descending and descending electrophoretic patterns are sufficiently non-enantiographic to suggest the existence of rapid, reversible interactions in the myogen solutions. However, no evidence of pronounced macromolecular association was obtained in velocity-sedimentation experiments. The source of the non-enantiography in electrophoresis has been traced to interactions of phosphate with components of myogen, which should therefore be considered as a mixutre, rather than a complex, of glycolytic enzymes.


The binding of echinomycin to deoxyribonucleic acid. Echinomycin is a peptide antibiotic which binds strongly to double-helical DNA up to a limit of approximately one molecule per five base-pairs. There is no detectable interaction with rRNA and only extremely feeble non-specific interaction with poly(rA)-poly(rU). Heat denaturation of DNA greatly decreases the binding, and similarly limited interaction is observed with naturally occurring single-stranded DNA. Association constants for binding to nine double-helical DNA species from different sources are presented; they vary by a factor of approximately 10, but are not simply related to the gross base composition. The interaction with DNA is ionic-strength-dependent, the binding constant falling by a factor of 4 when the ionic strength is raised from 0.01 to 0.10mol/litre. From the effect of temperature on the association constant for calf thymus DNA, the enthalpy of interaction is calculated to be about -13kJ/mol (-3kcal/mol). Binding of echinomycin persists in CsCl gradients and the buoyant density of nicked bacteriophage PM2 DNA is decreased by 25 mg/ml. Echinomycin interacts strongly with certain synthetic poly-deoxynucleotides, the binding constant decreasing in the order poly(dG)-poly(dC) greater than poly(dG-dC) greater than poly(dA-dT). For the latter two polymers the number of base-pairs occluded per bound antibiotic molecule is calculated to be three, whereas for poly(dG)-poly(dC) it is estimated to be four to five. Poly(dA)-poly(dT) and poly(dI)-poly(dC) interact only very weakly with the antibiotic. Poly(dI-dC) interacts to a slightly greater extent, but the binding curve is quite unlike that seen with the three strongly binding synthetic polynucleotides. Echinomycin affects the supercoiling of closed circular duplex bacteriophage PM2 DNA in the characteristic fashion of intercalating drugs. At low ionic strength the unwinding angle is almost twice that of ethidium. Likewise the extension of the helix, determined from changes in the viscosity of rod-like sonicated DNA fragments, is nearly double that expected for a simple (monofunctional) intercalation process. On this basis the interaction process is characterized as bifunctional intercalation. At higher ionic strength the unwinding angle relative to that of ethidium and the helix extension per bound echinomycin molecule fall, indicating a smooth progression towards more nearly monofunctional intercalation. Two simpler compounds which act as analogues of the quinoxaline chromophores of echinomycin, quinoxaline-2-carboxamide and the trypanocidal drug Bayer 7602, interact with DNA very much more weakly than does echinomycin, showing that the peptide portion of the antibiotic plays an essential role in determining the strength and specificity of the interaction.


The occurrence of cis-octadec-15-enoic acid as a major biohydrogenation product from methyl linolenate in bovine rumen liquor. The biohydrogenation products from methyl linolenate varied according to its concentration in the rumen. At low concentrations, over 60% of the biohydrogenated products was stearic acid, but when the concentration was raised above 100 mg/100 ml of rumen liquor, monoenoic acids, including cis-octadec-15-enoic acid, were the major products.


The chemical constitution of the proteoglycan of human intervertebral disc. Proteoglycan was prepared from three pools of normal human intervertebral discs by extraction with buffered 4M-guanidinium chloride followed by CsCl-density-gradient ultracentrifugation. Chromatography on agarose (Bio-Gel A-150m) and on DEAE-cellulose suggested a single polydisperse proteoglycan species. The intrinsic viscosities of three preparations were 166, 122 and 168 ml/g. After degradation with 0.5M-KOH containing 0.02M-NaBH4, the glycosaminoglycans were recovered quantitatively and their Ca2+ salts separated into a hexuronate-rich fraction (fraction 1), which was precipitated in 0-45% (v/v) ethanol, and a hexose-rich fraction (fraction2), which was precipitated in 45-70% (v/v) ethanol. Qualitative and quantitative analyses of the glycosaminoglycans revealed fraction 1 to be chondroitin sulphate, and fraction 2 to be keratan sulphate; the latter was contaminated with protein and possibly a small amount of another glycosaminoglycan. For both glycosaminoglycans, plots of log(mol.wt.) against weight fell close to a normal distribution. The mode for chondroitin sulphate was close to 20000; that for keratan sulphate, 10000. A threefold range of molecular weight included the central 16-84% [+/- 1 S.D. of log(mol.wt.)] of the weight of both fractions.


The subunit structure of thyroglobulin.

The subunit structure of thyroglobulin. Human and rat thyroglobulin were reduced and alkylated in aqueous alkaline conditions in the absence of denaturants; the product of reduction in both cases has been found to have mol.wt. about 165000, or one-quarter that of the native molecule.


A new type of representation of dipeptide conformation. The possibility of representing the conformations of a pair of peptide units in terms of two parameters, namely angle (nu) between the two peptide planes and the virtual-bond angle delta (C(alpha)i-1, C(alpha)i, C(alpha)i+1), and its usefulness in recognizing unfavourable conformations, is discussed. To exemplify the concept, the local conformations in lysozyme have been plotted in the (nu,delta) plane.


The separation and sequencing of permethylated peptides by mass spectrometry directly coupled to gas-liquid chromatography. The use of g.l.c. coupled to mass spectrometry to separate and sequence permethylated acetyl- and trifluoroacetyl-peptides in a single operation is described. Both electron impact and chemical ionization were used to induce fragmentation, and the latter was found to be more sensitive. Chromatographic retention data are presented which suggest that peptide derivatives of molecular weight of at least 750 are accessible to the technique. The application of our methods to the determination of the primary sequence of proteins is discussed.


Activation by cholera toxin of adenylate cyclase solubilized from rat liver. Cholera toxin, or peptide A1 from the toxin, activates adenylate cyclase solubilized from rat liver with Lubrol PX, provided that cell sap, NAD+, ATP and thiol-group-containing compounds are present. The activation is abolished by antisera to whole toxin, but not to subunit B.


Response of poly(adenylic acid) polymerase in rat liver nuclei and mitochondria to stravation and re-feeding with amino acids. Poly(adenylic acid) polymerase was extracted from liver nuclei and mitochondria of rats either fed ad libitum, starved overnight or starved and then re-fed with a complete amino acid mixture for 1-3 h. The enzymes were partially purified and assayed by using exogenous primers. Starvation resulted in an 80% decrease in the total activity of the purified nuclear enzyme, and the mitochondrial enzyme activity diminished to almost zero after overnight starvation. Measurements of the protein content of whole nuclei or mitochondria and of the enzyme extracts from these organelles indicated that the decrease in enzyme activity on starvation was not caused by incomplete extraction of the enzyme from the starved animals. Re-feeding the animals with the complete amino acid mixture increased the total activity of poly(A) polymerase from the nuclei and mitochondria by 1.9-fold and 63-fold respectively. Under these conditions, the total protein content of the nuclei and mitochondria increased by only 13 and 32% respectively. These data indicate that poly(A) polymerase is one of the cellular proteins specifically regulated by amino acid supply.


Effect of 2-allyl-2-isopropylacetamide on poly(adenylic acid)-containing ribonucleic acid. A single administration of 2-allyl-2-isopropylacetamide, a porphyrinogenic drug, enhanced the 32P-labelling of nucleoplasmic as well as cytoplasmic poly(A)-containing RNA in rat liver. The synthesis of total microsomal RNA is only marginally increased under these conditions. The drug enhances the labelling of a variety of cytoplasmic poly(A)-containing RNA species, and this effect is counteracted by the simultaneous administration of haemin. 2-Allyl-2-isopropylacetamide also enhanced the release of RNA from the nucleus to the cytoplasm.


The effect of glucose, insulin and adrenaline on glycerol metabolism in vitro in rat adipose tissue. The uptake and utilization of [1-14C]glycerol was determined in pieces of rat epididymal fat-pads incubated in Krebs--Ringer bicarbonate buffer containing albumin. Insulin (200 muunits/ml), adrenaline (epinephrine; 0.5 mug/ml) and glucose (0, 5, 15 and 20 mM) were added to the medium. Changes in the specific radioactivity of the tracer during the incubation were taken into account in calculating the rate of glycerol utilization. Adrenaline decreased glycerol uptake, whereas insulin plus adrenaline increased it. The rate of incorporation of glycerol into glycerides was decreased by adrenaline and insulin, singly or together. Insulin increased the rate of formation of CO2 and fatty acids from glycerol. The formation of CO2 and fatty acids was further enhanced by insulin plus adrenaline. The decrease in glycerol uptake induced by adrenaline, the decrease in incorporation of glycerol into glycerides induced by insulin and insulin plus adrenaline and the synthesis of fatty acids were dependent on the presence of glucose in the medium. Thus insulin and adrenaline act on glycerol utilization in adipose tissue and some of their effects are mediated by action on glucose metabolism, but others are independent of this.


The biochemical pathway for the breakdown of methyl cyanide (acetonitrile) in bacteria. [2-14C]Methyl cyanide (acetonitrile) is metabolized to citrate, succinate, fumarate, malate, glutamate, pyrrolidonecarboxylic acid and aspartate. Non-radioactive acetamide and acetate compete with 14C from methyl cyanide, and [2-14C]acetate and [2-14C]methyl cyanide are metabolized at similar rates, giving identical products. This evidence, combined with the inhibitory effect of fluoroacetate and arsenite on methyl cyanide metabolism, indicates that the pathway is: methyl cyanide leads to acetamide leads to acetate leads to tricarboxylic acid-cycle intermediates. The pathway was investigated in a species of Pseudomonas (group III; N.C.I.B. 10477), but comparison of labelling patterns suggests that it also exists in several higher plants.


Uptake of mevalonate and acetate during plastid development. 1. The envelopes of etioplasts and 1-2h etiochloroplasts are permeable to mevalonate, but plastids from etiolated tissue illuminated for longer than 4h show progressive impermeability towards mevalonate. 2. Acetate permeates the envelopes of 1-4h etiochloroplasts but does not significantly cross the envelopes of etioplasts or 8-24h etiochloroplasts. 3. A translocator system exists within the plastid envelopes for mevalonate which relies on malate as a counter-exchange anion.


Comparison of renal and hepatic glutathione S-transferases in the rat. Renal and hepatic GSH (reduced glutathione) S-transferase were compared with respect to substrate and inhibitory kinetics and hormonal influences in vivo. An example of each of five classes of substrates (aryl, aralkyl, epoxide, alkyl and alkene) was used. In the gel filtration of renal or hepatic cytosol, an identical elution volume was found for all the transferase activities. Close correspondence in Km values was found for aryl, epoxide- and alkyl-transferase activities, with only the aralkyl activity significantly lower in kidney. Probenecid and p-aminohippurate were competitive inhibitors of renal aryl-, aralkyl-, epoxide- and alkyl-transferase activities and inhibited renal alkene activity. Close correspondence in Ki values for inhibition by probenecid of these activities in kidney and liver was found. In addition, furosemide was a potent competitive inhibitor of renal alkyl-transferase activity. Hypophysectomy resulted in significant increases in aryl-, araklyl-, and expoxide-transferase activities in liver and kidney. The hypophysectomy-induced increases in renal aryl- and aralkyl-transferase activities (approx. 100%) were more than twofold greater than increases in hepatic activities (approx. 40%). Administration of thyroxine prevented the hypophysectomy-induced increase in aryltransferase activity in both kidney and liver. The renal GSH S-transferases, in view of similarities to the hepatic activities, may play a role as cytoplasmic organic-anion receptors, as previously proposed for the hepatic enzymes.


Phosphatidic acid biosynthesis in rat liver mitochondria and microsomal fractions. Regulation of fatty acid positional specificity. The positional and fatty acid specificity of phosphatidic acid biosynthesis in rat liver mitochondria and microsomal fractions was studied by using acylcarnitines, CoA and an excess of carnitine palmitoyltransferase (EC 2.3.1.21) as the source of acyl-CoA. In the mitochondria, the preference for palmitic acid at the 1-position is increased at high acyl-CoA concentrations, whereas it is decreased in the microsomal fraction. There was no change in the fatty acid specificity at the 2-position with different acyl-CoA concentrations in any of the factions. The preference in mitochondria for linoleic acid at the 2-position is strongly increased at high concentrations of lysophosphatidic acid.


Androgen-dependent synthesis of basic secretory proteins by the rat seminal vesicle. 1. Two basic proteins were purified from secretions of rat seminal vesicles by using Sephadex G-200 chromatography and polyacrylamide-gel electrophoresis under denaturing conditions. 2. It is not certain that these two proteins are distinct species and not subunits of a larger protein, but their properties are similar. Highly basic (pI = 9.7), they migrate to the cathode at high pH and their amino acid composition shows them to be rich in basic residues and serine. Threonine and hydrophobic residues are few. Both proteins are glycoproteins and have mol.wts. of 17000 and 18500. 3. Together these two proteins account for 25-30% of the protein synthesized by the vesicles, but they are absent from other tissues. 4. Changes in androgen status of the animal markedly affect these proteins. After castration, a progressive decrease in the basic proteins is observed and the synthesis of the two proteins as measured by [35S]methionine incorporation in vitro is is decreased. Testosterone administration in vivo rapidly restores their rates of synthesis. 5. These effects on specific protein synthesis are also observed for total cellular protein, and it is suggested that testosterone acts generally on the total protein-synthetic capacity of the cell and not specifically on individual proteins. Proliferative responses in the secretory epithelium may also be involved. 6. The extreme steroid specificity of the induction process suggests that the synthesis of these basic proteins is mediated by the androgen-receptor system. 7. The biological function of these proteins is not clear, but they do not appear to be involved in the formation of the copulatory plug.


Arachidonic acid release from diacyl phosphatidylethanolamine by human platelet membranes. At pH9.5 in the presence of 10 mM-Ca2+, human platelet membranes released 22% (167 of 785 nmol) of arachidonic acid that was esterified to phospholipids. With the use of synthetic choline (dinonadecanoyl) and ethanolamine (diheptadecanoyl) phosphoglycerides as internal reference compounds, 115 nmol of the released arachidonic acid was shown to be derived from endogenous breakdown of the phosphatidylethanolamine fraction. Further, the lysophosphatidylethanolamine that was released along with the arachidonic acid was shown virtually to lack fatty aldehydes and to contain a preponderance of fatty acids that have a preference for esterification at the 1-position of naturally occurring phosphatidylethanolamine of human platelets. These findings ruled out plasmalogen phosphatidylethanolamine as the source of the released arachidonic acid. We conclude that diacyl phosphatidylethanolamine was the principal source of arachidonic acid released by human platelet membranes under the conditions described.


The effect of lead on the calcium-handling capacity of rat heart mitochondria. 1. Very low concentrations of Pb2+ decrease the capacity of rat heart mitochondria, oxidizing pyruvate plus malate, to remove Ca2+ from the medium. 2. The primary effect is on the rate of Ca2+ sequestration; this is reflected in the overall extent of Ca2+ removal. 3. Pb2+ has at least two separate actions. Below about 0.5 nmol/mg of protein, it acts solely by competing with Ca2+ (Ki = 0.4 muM); above this concentration it also inhibits the production or use of respiratory energy, so that at 1 nmol of Pb2+/mg of protein, Ca2+ removal is almost completely abolished. 4. Pb2+ inhibits coupled and uncoupled respiratory O2 use by mitochondria oxidizing pyruvate plus malate, but at higher concentrations than those that affect Ca2+ removal; similar concentrations of Pb2+ inhibit pyruvate uptake, but not malate uptake, by the mitochondria. 5. Mg2+ only decreases Ca2+ removal by competition, and is a far-less effective competitor than Pb2+ (Ki = 0.15 mM). It is possible that the primary cause of the second effect of Pb2+ is displacement of membrane Mg2+. 6. The consequences of these results are discussed in terms of the possible involvement of heart mitochondria in excitation-contraction coupling, and the Pb2+ levels that might occur in heart tissue in vivo.


A kinetic analysis of the changes in fluorescence on the interaction of 8-anilinonaphthalene-1-sulphonate with submitochondrial particles. A comparison of the fluorescence change on the addition of 8-anilinonaphthalene-1-sulphonate to succinate-energized submitochondrial particles with that on the addition of succinate to submitochondrial particles incubated with 8-anilinonaphthalene-1-sulphonate shows that these changes in fluorescence may be explained solely in terms of 8-anilinonaphthalene-1-sulphonate binding. This comparison does not support the proposal of an 8-anilinonaphthalene-1-sulphonate-monitored change in the conformation of submitochondrial-particle membranes [Brocklehurst, Freedman, Hancock & Radda (1970) Biochem. J.116, 721-731]. The biphasic nature of the decrease in fluorescence, which was found to follow the addition of uncoupler to submitochondrial particles incubated with ATP or succinate, or of antimycin A to submitochondrial particles incubated with succinate, does not support the existence of 'aplectic' and 'symplectic' states of the mitochondrial membrane [Barrett-Bee & Radda (1972) Biochim, Biophys. Acta 267, 211-215].


The effects of diphenyleneiodonium on mitochondrial reactions. Relation of binding of diphenylene[125I]iodonium to mitochondria to the extent of inhibition of oxygen uptake. 1. Several ring-substituted derivatives of diphenyleneiodonium catalyse the exchange of Cl- and OH- ions across the inner membrane of rat liver mitochondria. They also inhibit state 3 and state 3u oxidations of glutamate plus malate in the presence of Cl- more than in its absence. Most have activities similar to diphenyleneiodonium, although 2,4-dichlorodiphenyleneiodonium is up to 50 times more active. 2. Diphenyleneiodonium inhibits soluble rat liver NADH dehydrogenase and NADH oxidation by rat liver sub-mitochondrial particles directly; 2,4-dichlorodiphenyleneiodonium is only about twice as inhibitory. 3. Liver mitochondria contain two classes of binding sites for diphenylene[125I]iodonium, namely high-affinity sites with an affinity constant of 3 X 10(5) M-1 (1--2 nmol/mg of protein), and low-affinity sites with an affinity constant of 1.3 X 10(3) M-1 (80 nmol/mg of protein). Both sites occur in hepatocytes with a relative enrichment of the low-affinity site. Nadh dehydrogenase preparations only apparently contain high-affinity binding sites. Only low-affinity sites occur in erythrocytes. 4. 2,4-Dichlorodiphenyleneiodonium competes with diphenylene[125I]iodonium for both low- and high-affinity sites, whereas tri-n-propyltin only competes for the low-affinity sites. 5. The high-affinity sites are apparently associated with NADH dehydrogenase and the low-affinity sites probably represent electrostatic binding of diphenylene[125I]iodonium to phospholipids. The high-affinity site does not appear to be associated with a rate-limiting stage of NADH oxidation.


The regulation of folate and methionine metabolism. 1. The isolated perfused rat liver and suspensions of isolated rat hepatocytes fail to form glucose from histidine, in contrast with the liver in vivo. Both rat liver preparations readily metabolize histidine. The main end product is N-formiminoglutamate. In this respect the liver preparations behave like the liver of cobalamin- or folate-deficient mammals. 2. Additions of L-methionine in physiological concentrations (or of ethionine [2-amino-4-(ethylthio)butyric acid]) promotes the degradation of formiminoglutamate, as is already known to be the case in cobalamin of folate deficiency. Added methionine also promotes glucose formation from histidine. 3. Addition of methionine accelerates the oxidation of formate to bicarbonate by hepatocytes. 4. A feature common to cobalamin-deficient liver and the isolated liver preparations is taken to be a low tissue methionine concentration, to be expected in cobalamin deficiency through a decreased synthesis of methionine and caused in liver preparations by a washing out of amino acids during the handling of the tissue. 5. The available evidence is in accordance with the assumption that methionine does not directly increase the catalytic capacity of formyltetrahydrofolate dehydrogenase; rather, that an increased methionine concentration raises the concentration of S-adenosylmethionine, thus leading to the inhibition of methylenetetrahydrofolate reductase activity [Kutzbach & Stokstad (1967) Biochim. Biophys. Acta 139, 217-220; Kutzbach & Stokstad (1971) Methods Enzymol. 18B, 793-798], that this inhibition causes an increase in the concentration of methylenetetrahydrofolate and the C1 tetrahydrofolate derivatives in equilibrium with methylenetetrahydrofolate, including 10-formyltetrahydrofolate; that the increased concentration of the latter accelerates the formyltetrahydrofolate dehydrogenase reaction, because the normal concentration of the substrate is far below the Km value of the enzyme for the substrate. 6. The findings are relevant to the understanding of the regulation of both folate and methionine metabolism. When the methionine concentration is low, C1 units are preserved by the decreased activity of formyltetrahydrofolate dehydrogenase and are utilized for the synthesis of methionine, purines and pyrimidines. On the other hand when the concentration of methionine, and hence adenosylmethionine, is high and there is a surplus of C1 units as a result of excess of dietary supply, formyltetrahydrofolate dehydrogenase disposes of the excess. When ample dietary supply causes an excess of methionine, which has to be disposed of by degradation, the increased activity of formyltetrahydrofolate dehydrogenase decreases the supply of methyltetrahydrofolate. Thus homocysteine, instead of being remethylated, enters the pathway of degradation via cystathionine. 7. The findings throw light on the biochemical abnormalities associated with cobalamin deficiency (megaloblastic anaemia), especially on the 'methylfolate-trap hypothesis'. This is discussed. 8...


Effects of hormones on protein and amino acid metabolism in mammary-gland explants of mice. The effects of insulin, cortisol and prolactin on amino acid uptake and protein biosynthesis were determined in mammary-gland explants from mid-pregnant mice. Insulin stimulated [3H]leucine incorporation into protein within 15 min of adding insulin to the incubation medium. Insulin also had a rapid stimulatory effect on the rate of aminoiso[14C]butyric acid uptake, but it had no effect on the intracellular accumulation of [3H]leucine. Cortisol inhibited the rate of [3H]leucine incorporation into protein during the initial 4h of incubation, but it had no effect at subsequent times. [3H]Leucine uptake was unaffected by cortisol, but amino[14C]isobutyric acid uptake was inhibited after a 4h exposure period to this hormone. Prolactin stimulated the rate of [3H]leucine incorporation into protein when tissues were exposed to this hormone for 4h or more; up to 4h, however, no effect of prolactin was detected. At all times tested, prolactin had no effect on the uptake of either amino[14C]isobutyric acid or [3H]leucine. Incubation with actinomycin D abolished the prolactin stimulation of protein biosynthesis, but this antibiotic did not affect the insulin response. A distinct difference in the mechanism of action of these hormones on protein biosynthesis in the mammary gland is thus apparent.


Studies on the relative biopotencies and intestinal absorption of different apo-beta-carotenoids in rats and chickens. 1. The biopotencies relative to beta-carotene of several apocarotenoids, such as 8'-, 10'- and 12'-apo-beta-carotenal and methyl 8'-apo-beta-carotenoate, were investigated in rats, on a molar basis, by both curative-growth assay and liver-storage tests. 2. In the curative-growth assays, on a molar basis the biopotencies of 8'-, 10'- and 12'-apo-beta-carotenal and methyl 8'-apo-beta-carotenoate were 72, 78, 72 and 53% respectively, whereas on a weight basis the corresponding values were 93, 111, 111 and 63%, with respect to beta-carotene taken as 100%. In terms of yield of vitamin A, these values were much lower in the liver-storage tests. 3. When 8'-apo-beta-carotenal was fed, the unchanged aldehyde together with small amounts of the corresponding alcohol and larger proportions of the acid rapidly appeared in the tissues of both rats and chickens. The 8'-apocarotenol, 8'-apocarotenoic acid and its methyl ester were absorbed unchanged. The free acid disappeared most rapidly from the tissues, but its methyl ester persisted in the tissues longest. 4. On the basis of these observations it is suggested that most of an apocarotenal is oxidized to the corresponding acid, which, in turn, is mostly degraded to retinoic acid, with small proportions of it being attacked by the dioxygenase system giving retinal.


Role of protein degradation in the growth of livers after a nutritional shift. Fractional rates of synthesis and degradation of liver porteins were estimated during the rapid restoration of liver mass observed in protein-depleted mice when they are fed with an adequate diet. 1. Net protein gain was fastest 12h after the nutritional shift, when it reached a rate of 48% per day. 2. The RNA/protein ratio in livers of protein-depleted animals was essentially the same as in normal livers; it increased by a maximum of 13% 12h after the nutritional shift. 3. Rates of protein synthesis in vivo were measured by the incorporation into liver protein of massive amounts of L-[1-14C]leucine. In protein-depleted animals, the rate of synthesis per mg of RNA was 72% of that in normal livers. Normal rates were recovered within 12h of the nutritional shift. 4. The fraction of newly synthesized protein retained by the liver was studied after they were pulse-labelled by the intravenous injection of radioactive leucine, and, 5 min later, pactamycin (an inhibitor of the initiation of protein synthesis); 3h later the livers in both experimental situations retained 58% of the newly synthesized protein. 5. Fractional rates of protein degradation were estimated either from the difference between the synthesis of stable liver proteins and the net protein increase, or by the disappearance of radioactivity from the liver protein previously labelled by the administration to the mice of NaH14CO3. Both procedures demonstrated a large decrease in the rate of protein degradation during liver growth.


Synthesis of delta-aminolaevulinate synthase by isolated liver polyribosomes. 1. Postmitochondrial supernatants were prepared from the livers of chick embryos and were incubated under conditions that supported protein synthesis. delta-Aminolaevulinate synthase (EC 2.3.1.37) was synthesized by supernatants from livers treated with the porphyrinogenic drugs 2-allyl-2-isopropylacetamide and/or 3,5-diethoxycarbonyl-1,4-dihydrocollidine, but synthesis by supernatants from normal livers could not be detected. Synthesis of enzyme released from polyribosomes was measured by immunoprecipitation with specific antibody to the mitochondrial enzyme, and the specificity of the reaction was established by electrophoresis of dissociated immunoprecipitates on sodium dodecyl sulphate/polyacrylamide gels. 2. The relative synthesis of delta-aminolaevulinate synthase in vitro was comparable with that previously measured in vivo, and was correlated with the enzyme activity of the liver. 3. Enzyme synthesis in vitro occurred predominantly on free rather than membrane-bound polyribosomes. 4. The mol.wt. of the product synthesized in vitro was 7000 +/- 7000 by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. However, pulse-labelling of the enzyme in vivo confirmed its mol.wt. to be 49000 +/- 5000 when isolated from the mitochondrion. A small amount of immunoprecipitable enzyme of mol.wt. 70000 was detected in the cytosol in vivo. In chick embryo liver, delta-aminolaevulinate synthase therefore appears to be synthesized on cytoplasmic polyribosomes as a polypeptide of mol.wt. 70000, which in vivo is rapidly incorporated into the mitochondrion, and is then extracted as a lower-molecular-weight form. 5. Haemin added to the postmitochondrial supernatant-containing incubation mixture at concentrations up to 10 muM had no effect on general protein synthesis or the synthesis of delta-aminolaevulinate synthase. On the other hand, haemin treatment of induced chick embryo livers in vivo for 3h markedly decreased the relative synthesis of delta-aminolaevulinate synthase in vitro. These results suggest that haemin represses the synthesis of delta-aminolaevulinate synthase by decreasing the amount of mRNA for the enzyme available for translation.


Liposomal retention of a modified anti-inflammatory steroid. In studies with synthetic lecithins, dipalmitoylphosphatidylcholine was found to be the preferred form for liposomal retention of cortisol esters at 37 degrees C. Cortisol palmitate was retained longer than cortisol octanoate, whereas unesterified cortisol escaped readily from liposomes. Such a liposome composition may allow the controlled release of modified anti-inflammatory agents, particularly when used for intra-articular administration.


Ribonucleic acid synthesis in the renal cortex at the initiation of compensatory growth. The mechanisms responsible for the increase in RNA per cell during the first 48h of renal compensatory growth were studied in the renal cortex. Unilaterally nephrectomized, sham-operated or non-operated rats were used. Incorporation into RNA of labelled precursors was studied in vivo and in vitro. Sham-operation produced significant changes in precursor incorporation, absolute amounts of UTP and RNA, and the rate of RNA  synthesis. At 6h after surgery, the amount of RNA decreased in sham-operated controls, whereas that in growing cortex remained unchanged. Incorporation into RNA in vivo was greater in the growing cortex, although the rate of RNA synthesis was not increased. At 24h, precursor incorporation into RNA and UTP and RNA synthesis were all increased in the growing cortex. In contrast with results obtained in vivo, slices of growing cortex incorporated less labelled precursor into RNA than did cortex slices from sham-operated controls, from 3 to 48h. Maximal differences were found from 6 to 24h. An attempt was made to equalize endogenous precursor pool sizes by increasing the concentration of unlabelled uridine in the media; incorporation differences were narrowed significantly. Serum from nephrectomized animals did not increase precursor incorporation into RNA in vitro. An increase in RNA synthesis is an important factor in RNA accretion in the renal cortex beyond 12h of compensatory growth. This is accompanied by increased UTP content and preceded by expansion of other pools. The amount of labelled precursor incorporated into RNA is greatly influenced by its delivery rate to the growing kidney in vivo and by intracellular dilution of expanded precursor pools in vitro.


Localization of thyroid peroxidase and the site of iodination in rat thyroid gland. The iodinated protein was localized in thyroid tissue slices by using radioautography. In unfixed tissue, the labelled protein was localized in the colloid, whereas, in tissue that was fixed before the 125I addition, the label was within the follicular cell. This localizes thyroid peroxidase largely on the endoplasmic reticulum of the cell.


Mammary glucocorticoid receptor of mice in pregnancy and in lactation. Activity of glucocorticoid receptor in mouse mammary cytosol changes during lactogenesis. The highest receptor activity is observed in the second half of pregnancy. The receptor from mammary glands from lactating and pregnant mice exhibits the same sedimentation pattern, as well as the same specificity and affinity for binding of steroid hormones.


Inhibition of polyamine accumulation and deoxyribonucleic acid synthesis in regenerating rat liver. Repeated injections of 1,3-diaminopropane into rats after partial hepatectomy caused a repression-type inhibiton of liver ornithine decarboxylase (EC 4.1.1.17) and totally prevented the marked increases in liver putrescine and spermidine concentrations that normally occur in response to partial hepatectomy. The inhibition of polyamine synthesis by diaminopropane was accompanied by a profound decrease (about 80%) in the synthesis of DNA in the regenerating rat liver without any changes in the synthesis of RNA and total liver protein.


Failure of a cell-free system from rabbit reticulocytes to remove iron from transferrin. 1. Cell-free solutions prepared from rabbit reticulocytes were not able to release iron from rabbit transferrin. 2. The results, which differ from those obtained with Rana catesbeiana immature erythrocytes, indicate that cellular integrity is a requirement for this process in the rabbit reticulocyte system.


Identification of a sialoglycopeptide released by self-digestion from human erythrocyte membranes. Membranes from human O Rhesus-positive erythrocyte 'ghosts' were tested in vitro for their ability to digest their own glycoproteins. 'Ghost' membranes incubated in Tris/HCl buffer, pH 7.4, release a sialoglycopeptide, which contains glucosamine, galactosamine, galactose and mainly polar amino acids. Chemical composition, molecular size and aggregation properties suggest that this glycopeptide may be a fragment of glycophorin.


Leucine and D-3-hydroxybutyrate as lipid precursors in developing rat spinal cord and liver. By using a labelled ketone body, D-3-hydroxy[3-14C]butyrate, or a ketone-body precursor, L-[4,5-3H]leucine, preferential labelling of spinal-cord cholesterol was demonstrated. In contrast, the phospholipid fraction was more heavily labelled in the liver. In both tissues phosphatidylcholine was the major labelled complex lipid. Incorporation ob both substrates into total lipid, on a tissue-weight basis, was appreciably greater for spinal cord.


Cholinergically stimulated phosphatidylinositol breakdown in parotid-gland fragments is independent of the ionic environment. Carbamoylcholine stimulated phosphatidylinositol breakdown in rat parotid-gland fragments incubated in either a Krebs--Ringer medium or an ion-depleted iso-osmotic sucrose medium. This suggest that phosphatidylinositol breakdown is not initiated by a change in the intracellular concentration of Na+ or of K+, and previous experiments have already indicated that it is independent of cyclic nucleotides and Ca2+. Thus it seems that this reaction may be initiated by a direct interaction at the plasma membrane between the activated muscarinic cholinergic receptor system and the enzyme that catalyses phosphatidylinositol breakdown.


Neuronal N-acetyl-beta-D-glucosaminidase. Evidence for its biosynthesis in vitro. Neuronal cell bodies, isolated in bulk from 8-day-old rat cerebral cortices, were incubated in the presence of a 3H-labelled amino acid mixture, and subcellular fractions isolated by differential centrifugation. The particulate fractions were frozen/thawed in 0.20 M-sucrose/0.1 M-KCl [Selling et al. (1973) Biochim. Biophys. Acta 315, 128-146] and the profiles of acid-insoluble radioactivity and N-acetyl-beta-D-glucosaminidase (glucosaminidase) activity compared in the resulting non-sedimentable fractions by DEAE-cellulose chromatography and cellulose acetate electrophoresis. Radioactivity and glucosaminidase activity co-migrated to a significant extent. Electrophoresis revealed that after 1 min of incubation 42% of the radioactivity of the non-sedimentable microsomal fraction after freezing and thawing co-migrated with an intensely fluorescent band of glucosaminidase activity. Since the pellet fraction obtained on freezing/thawing the microsomal fraction contained up to 75% of the RNA, 95% of the radioactivity and 45% of the glucosaminidase, a detailed study of the association between its radioactivity and nascent glucosaminidase activity was undertaken. After 1 and 2 min of incubation, followed by centrifugation of the microsomal pellet on 35-60% (w/v) sucrose density gradients, radioactivity and glucosaminidase activity exhibited parallel profiles in the region of heavy polyribosomes and at the top of the gradient which contains spontaneously released nascent polypeptide chains. DEAE-cellulose chromatography of these chains revealed glucosaminidase A to be the principal nascent glucosaminidase component, with glucosaminidases B and C as minor peaks. After 2 min of incubation, all of the glucosaminidase components appeared labelled, and glucosaminidase A exhibited two distinct sub-components. The pattern of glucosaminidase labelling in the soluble and microsomal fractions suggested that newly formed glucosaminidase molecules traverse both the cellular sap and the lumen of the endoplasmic reticulum. Only glucosaminidase A reacted specifically with concanavalin A and radioactive glucosaminidase A could be successfully regenerated by treatment with alpha-methyl-D-glucoside. Glucosaminidase A and a substantial portion of the radioactivity associating with it could be readily converted into glucosaminidase B by re-chromatography on DEAE-cellulose and by reaction of the concanavalin A-glucosaminidase A complex with methyl glucosides.


Altered polyamine metabolism in the PRO/Re strain of inbred mice. The PRO/Re strain of inbred mice are characterized by abnormally high concentrations of proline in both blood (hyperprolinaemia) and urine (prolinuria). They excrete increased amounts of polyamines in their urine. Male PRO/Re mice excreted putrescine at 175% and spermidine at 300% the amount of male C57BL/6J controls. Female PRO/Re mice excreted putrescine at 115% and spermidine at 150% of the amount in the urine of female controls. Examination of the enzymes involved in polyamine biosynthesis revealed that ornithine decarboxylase, the initial enzyme in the polyamine-biosynthetic pathway, was increased by 150% in the kidneys and by 100% in the liver of male PRO/Re mice. There was no significant difference between PRO/Re and C57BL/6J male mice for either putrescine- or spermidine-stimulated S-adenosylmethionine decarboxylase activity. Female PRO/Re mice showed no significant difference from female C57BL/6J mice for any of the enzymes examined. When the concentrations of the polyamines in the tissues of the PRO/Re mice were determined, spermidine and spermine concentrations in the kidneys of the male PRO/Re mice were twice those of the controls. Spermidine concentration in the livers of both male and female PRO/Re mice was approx. 130% that of the controls. Polyamine concentrations in the brains were similar in controls and mutants. The increased polyamine biosynthesis and excretion in the PRO/Re mutant mice may be a mechanism to decrease the extent of proline accumulation.


The effects of added purines on urate and purine synthesis de novo by isolated chick liver, kidney and lymphoid cells. 1. Isolated chick lymphoid cells, together with isolated chick liver and kidney cells, incorporate [1-14C]glycine or [14C]formate into urate. 2. Of the cell types used, bursal cells incorporate 14C into urate at the fastest rate, although the output of total urate by bursal cells is only 10% that of liver cells. 3. When suspended in Eagle's medium the incorporation of 14C into urate is inhibited by adenine and guanine up to 1 mM. In contrast, the addition of 1 mM-AMP or -GMP results in a relatively large stimulation of this incorporation. 4. Added adenine is rapidly taken up by liver cells and then released in an unmetabolized form; AMP is taken up more slowly and is rapidly metabolized. The metabolites (possibly including adenine) are then released. 5. Intracellular liver 5-phosphoribosyl 1-pyrophosphate is approx. 0.7mM and remains constant or falls slightly during a 3 h incubation of the cells. 6. The addition of adenine or guanine, AMP or GMP, does not alter liver intracellular 5-phosphoribosyl 1-pyrophosphate concentrations. Added 5-phosphoribosyl 1-pyrophosphate is not taken up by liver cells. 7. The results are discussed in the context of the control of urate and purine synthesis de novo in the chick.


The synthesis of glycosaminoglycans by cultures of rabbit corneal endothelial and stromal cells. Confluent monolayer cultures of rabbit corneal endothelial and stromal cells were incubated independently with [35S]sulphate and [3H]glucosamine for 3 days. AFter incubation, labelled glycosaminoglycans were isolated from the growth medium and from a cellular fraction. These glycosaminoglycans were further characterized by DEAE-cellulose column chromatography and by sequential treatment with various glycosamino-glycan-degrading enzymes. Both endothelial and stromal cultures synthesized hyaluronic acid as the principal product. The cell fraction from the stromal cultures, however, had significantly less hyaluronic acid than that from the endothelial cultures. In addition, both types of cells synthesized a variety of sulphated glycosaminoglycans. The relative amounts of each sulphated glycosaminoglycan in the two cell lines were similar, with chondroitin 4-sulphate, chondroitin 6-sulphate and dermatan sulphate as the major components. Heparan sulphate was present in smaller amounts. Keratan sulphate was also identified, but only in very small amounts (1-3%). The presence of dermatan sulphate and the high content of hyaluronic acid are similar to the pattern of glycosaminoglycans seen in regenerating or developing tissues, including cornea.


Adenosine triphosphate-evoked catecholamine release in chromatin granules. Osmotic lysis as a consequence of proton translocation. Chromaffin granules suspended in C1-containing media release catecholamine and protein when ATP is added. This phenomenon is inhibited in hyperosmotic media and in the presence of uncouplers of oxidative phosphorylation. Release requires a permeant anion in the medium, but is independent of the cation. The release process appears to be driven by an inwardly directed proton-translocating adenosine triphosphatase. The resulting proton-anion influx causes osmotic lysis of the chromaffin granules.


Glucosamine metabolism in regenerating rat liver. 1. Glycoprotein synthesis was investigated with [1-14C]glucosamine in vivo. [14C]Glucosamine was administered intravenously 24h after hepatectomy to rats. 2. Incorporation into the acid-soluble fraction was maximum at 15 min after injection both in sham-operated and hepatectomized rats. 3. Enhancement of incorporation into UDP-N-acetylhexosamine in regenerating liver was observed. However, its specific activity was lower, because of a greater enhancement of synthesis de novo of the amino sugar. 4. In the liver acid-insoluble fraction, maximum incorporation of [14C]glucosamine was at 30 min in sham-operated rats and 2 h in hepatectomized rats respectively. 5. In sham-operated rats, incorporation into the plasma acid-insoluble fraction followed that of the liver acid-insoluble fraction, but hepatectomy resulted in a rapid enchancement of incorporation into plasma. 6. It is concluded that synthesis of liver glycoproteins is stimulated after partial hepatectomy and that glycoproteins synthesized are released rapidly into the plasma.


High rates of [1-14C]acetate incorporation into the lipid of isolated spinach chloroplasts. Spinach chloroplasts, isolated by techniques yielding preparations with high O2- evolving activity, showed rates of light-dependent acetate incorporation into lipids 3-4 fold higher than any previously reported. Incorporation  rates as high as 500 nmol of acetate/h per mg of chlorophyll were measured in buffered sorbitol solutions containing only NaHCO3 and [1-14C]acetate, and as high as 800 nmol/h per mg of chlorophyll when 0.13 mM-Triton X-100 was also included in the reaction media. The fatty acids synthesized were predominantly oleic (70-80% of the total fatty acid radioactivity) and palmitic (20-25%) with only minor amounts (1-5%) of linoleic acid. Linolenic acid synthesis was not detected in the system in vitro. Free fatty acids accounted for 70-90% of the radioactivity incorporated and the remainder was shared fairly evenly between 1,2-diacylglycerols and polar lipids. Oleic acid constituted 80-90% of the free fatty acids synthesized, but the diacylglycerols and polar lipids contained slightly more palmitic acid than oleic acid. Triton X-100 stimulated the synthesis of diacylglycerols 3-6 fold, but stimulated free fatty acid synthesis only 1-1.5-fold. Added glycerol 1-phosphate stimulated both the synthesis of diacylglycerols and palmitic acid relative to oleic acid, but did not increase acetate incorporation into total chloroplast lipids. CoA and ATP, when added separately, stimulated acetate incorporation into chloroplast lipids to variable extents and had no effect on the types of lipid synthesized, but when added together resulted in 34% of the incorporated acetate appearing in long-chain acyl-CoA. Pyruvate was a much less effective precursor of chloroplast fatty acids than was acetate.


Protein synthesis in salivary glands of Drosophila hydei after experimental gene induction. Several treatments, namely incubation at 37 degrees C, in the presence of arsenite, 2,4-dinitrophenol or vitamin B-6, or release from anaerobiosis induce the same set of puffs in the polythene chromosomes of salivary glands of Drosophila hydei. Analysis of changes in protein-synthetic patterns (as determined by radioautography of sodium dodecyl sulphate-gel electrophoretograms of extracts from [35S]methionine-labelled salivary glands) showed that concomitant with puff induction by these various treatments the same six strongly labelled polypeptide bands appeared. The amount of radioactive label in these peptides accounted for 25% of the total incorporation of [35S]methionine, except during incubation at 37 degrees C when it accounted for about 50%. The rate of synthesis of these peptides was maximal 1 h after the start of the puff-inducing treatment. The rate of decay of the rate of synthesis showed first-order kinetics both after removal of the puff-inducing stimulus or in the presence of actinomycin, with a half-life of approx. 4h.


The intracellular distribution of arylsulphatase C and oestrogen sulphatase activities in rat kidney and spleen. The arylsulphatase C of rat kidney and spleen is localized predominantly in the microsomal fraction. The distribution pattern is paralleled by that of oestrogen sulphatase, as in rat liver. The general usefulness of arylsulphatase C as a microsomal marker enzyme is indicated.


Selective resistance to desiccation of nuclear ribonucleic acid synthesis in isolated nuclei of Artemia salina embryos during pre-emergence development. The developing gastrula embryos of Artemia salina are resistant to a complete redesiccation during a period of pre-emergence development. Isolated nuclei from these dehydrated embryos could retain a transcriptional activity in vitro comparable with that of non-desiccated controls. On the other hand, redesiccation of both prenauplii and nauplii completely destroys their viability as well as the nuclear transcriptional activity. However, those gastrula embryos that did not develop in a first incubation period could remain viable and develop with a considerable time-lag after a subsequent second incubation.


Binding of diethylstilboestrol to deoxyribonucleic acid by rat liver microsomal fractions in vitro and in mouse foetal cells in culture. Diethylstillboestrol, a synthetic and carcinogenic hormone, binds to DNA as a result of incubation with a liver microsomal preparation in vitro and on incubation with primary mouse foetal cells in culture. Enzymic digestion of DNA samples thus prepared gives several covalent deoxyribonucleoside-diethylstilboestrol products from the microsomal system. One of these is produced in small but significant yield in the tissue-culture system.


Poly(ethylene glycol), surface potential and cell fusion. Poly(ethylene glycol), glycerol and dimethyl sulphoxide markedly decrease the surface potentials of monolayers of phosphatidylcholine and phosphatidylethanolamine. This finding is discussed in relation to the properties of hen erythrocytes undergoing fusion induced by poly(ethylene glycol).


Increase in sialic acids removable by neuraminidase during the shape change of platelets. Human or rabbit platelets were activated by ADP or 5-hydroxytrypatamine and rapidly fixed with glutaraldehyde. The shape change associated with activation gave rise to an increase in sialic acids removable by neuraminidase. This increase, like the shape change, was prevented by adenosine or methysergide added before ADP or 5-hydroxytryptamine respectively. The results indicate the exposure of additional glycoprotein(s) on the platelet surface.


Light- and electron-microscopic characteristics of artrial smooth muscle cell cultures subjected to hypoxia or carbon monoxide. Smooth muscle cells from the tunica media of piglet aortae grown under hypoxic conditions undergo the following changes: First, they become modified by partial loss of myofilaments and proliferation of organelles, which are characteristics of young primitive cells. Second, an increase in number of pinocytotic vesicles at and below the cell surface, indicating increased extracellular uptake of material, can be detected. This is followed by accumulation of Oil Red O positive intracytoplasmic granules and vacuoles as well as the subsequent formation of mount-like protrusions. The latter consist of a core of extracellular material and necrotic debris covered with a cap of viable cells. A third feature of the cells subjected to hypoxia is a conspicuous rise in the number of lysosomes. This is considered to be a manifestation of a defense mechanism of the cells to remove undesirable material from cytoplasm. Cells exposed to an atmosphere rich in carbon monoxide exhibit basically the same alterations as those grown under hypoxic conditions; however, formation of mound-like aggregates is less prominent, while the rise in the number of lysosomes is more evident than in the hypoxic cells. The above alterations are similar to changes observed in smooth muscle cells of rabbit with experimental atherosclerosis. It is suggested that whereever the arterial smooth muscle cell is subjected to adverse conditions basically the same mechanism, consisting of dedifferentiation, increased permeability and lysosomal defense reaction, takes place.


Effects of hypophysectomy on alloxan-diabetic, arteriosclerotic, breeder vs. non-arteriosclerotic, virgin rats. A single s.c. injection (10 mg/100 g bw of alloxan) was given to nonarteriosclerotic, virgin, Sprague--Dawley rats and to breeder rats with preexisting arteriosclerosis, hyperlipidemia and hyperglycemia. All of the animals promptly developed severe diabetes with ketosis, hyperglycemia, and hyperlipidemia. Insulin therapy was deliberately withheld. Mortality was high. Seven days later one group was subjected to hypophysectomy and 30 days later, all of the animals were autopsied. The diabetes + hypophysectomy animals maintained their body weight better, did not have hypertrophied adrenal glands, showed the least elevation of serum enzymes, e.g., CPK, SGOT, SGPT and LDH, less hyperlipidemia and hyperglycemia and reduced corticosterone production than the animals with untreated severe diabetes. Despite the relative amelioration of metabolic derangements prognostic of cardiovascular degenerative changes, the diabetes + hypophysectomy animals manifested extensive renovascular damage and the breeder rats with pre-existing arteriosclerosis showed definite exacerbation of their arterial disease in response to the severe alloxan diabetes regardless of hypophysectomy. It is suggested that although hypophysectomy may alleviate certain metabolic derangements attributed to growth hormone, ACTH and adrenal steroids, the angiopathic damage proceeds inexorably.


Susceptibility to experimental atherosclerosis: relation to mast cells and heparin. Fifteen chicken tissues were examined for mast cells. Numerous mast cells were found in peritoneum and spleen. The sulfated mucopolysaccharides extracted from these two tissues, corresponding in amount to that in mast cells, were found to be dermatan sulfate, chondroitin sulfate and heparitin sulfate but no evidence of heparin. We have shown a similar situation occurs in the rabbit which is also highly susceptible to the production of atherosclerosis by diet. These observations provide further evidence of a role for heparin and mast cells in limiting atherogenesis.


Composition of acidic glycosaminoglycans in human cerebral arteries. (1) The composition of acidic glycosaminoglycans (AGAG) in the pooled human cerebral arteries was investigated by electrophoretic characterization before and after digestion with chondroitinases. Constitution of the chondroitin sulfate (CS) family was determined qualitatively on the basis of the disaccharide subunits of the CS chains after depolymerization with the enzymes. (2) The data obtained indicated that the AGAG in cerebral arteries consisted of, in order of amount, heparan sulfates, chondroitin-6-sulfate, dermatan sulfate, chondroitin-4-sulfate and hyaluronic acid. (3) The existence of oversulfated CS and undersulfated CS in the cerebral AGAG was supported by the detection of unsaturated di-sulfated and non-sulfated disaccharides on paper chromatography. In addition, the presence of hyaluronic acid was indicated by electrophoretic and enzymatic separation. (4) The distribution of the individual AGAG in cerebral arteries was also examined on the basis of molecular weight by gel filtration on a Sephadex G-100 column.


Radio and microwave radiation and experimental atherosclerosis. The effect of radio and microwave radiation on dietary atherosclerosis of rabbits was tested. 16 New Zealand white rabbits were exposed to microwave (2.45 GHz) radiation at a power density of 20--30 mW/cm2 for 4 h a day, 5 days a week for 8 to 10 weeks. Irradiated animals had serum cholesterol concentrations, aortic wall cholesterol concentrations and percentage of intimal surface involved in atherosclerotic lesions which were not different from age and weight matched controls. Continuous radio frequency irradiation (1 MHz) for 8 to 11 weeks with a field strength of 30 V/cm also failed to change these indices of atherogenesis. We conclude that under the conditions of these experiments radio and microwave irradiation do not affect the course of diet induced atherogenesis.


Ascorbic acid metabolism in rats fed high fat cholesterol diet. The synthesis and catabolism of ascorbic acid has been studied in rats fed an atherogenic diet as also has the relation between the concentration of ascorbic acid and concentration of lipids in the tissues. The concentration of ascorbic acid was found to decrease in the serum, liver, spleen and adrenals, but not in the kidney, in the animals fed an atherogenic diet when compared with those fed normal diet. An inverse relationship was found between the concentration of ascorbic acid in the tissue and the concentration of lipids. The synthesis of ascorbic acid from D-glucuronalactone did not significantly differ in the liver and kidney, but decreased in the spleen in the animals fed the atherogenic diet. On the other hand, the catabolism of ascorbic acid significantly increased in the liver and spleen.


Emotional history and pathogenesis of cancer. Emotional history was evaluated in a sample of cancer patients and a sample of noncancer patients who were hospitalized for treatment. An in-depth interview was conducted with each S to identify the frequency, duration, and intensity of each emotion-provoking event that he could remember across his life history. Evaluations of the interview information identified more frequent and intense emotional events in the Ss with cancer than in the controls who did not have cancer.


Combination effects in chemical carcinogenesis (experimental results). Most of the malignant tumors of man are not caused by only one specific carcinogen, but have to be attributed to additive effects of various carcinogenic factors, predominantly from the environment. Experimental studies in rats and mice showed syncarcinogenic effects of small doses of different carcinogens with the same organotropy, given either simultaneously or consecutively. These effects were proved in both local and systemic carcinogens. Extensive studies carried out in some thousand rats showed that the action of pre-or postnatally applied chemical carcinogens is not influenced by immunostimulating or immunodepressive measures. In view of the topicality of combination effects of chemical carcinogens and lacking influences by regulative mechanisms of the host organism, it is recommended to eliminate chemical carcinogens as far as possible from the human environment.


Growth hormone's role in diabetic microangiopathy. The state of the blood vessels is normal at the clinical onset of juvenile diabetes. Vascular changes develop slowly and progressively. According to the growth hormone hypothesis, elevated serum growth hormone is one casual factor in the development of diabetic angiopathy. The hypothesis proposes an effect of growth hormone not on blood glucose but directly on blood vessels. This hypothesis is based on serum growth hormone studies and on a controlled clinical trial of the effect of hypophysectomy on small blood vessels. An animal model of large-vessel disease in diabetes is briefly described. There is a large molecule in diabetic serum causing proliferation of aortic myomedial cells in culture. Growth hormone causes a similar proliferation. A short summary is given of the present situation in somatostatin research relating to diabetes mellitus.


Regulation of renin release and intrarenal formation of angiotensin. Studies in the isolated perfused rat kidney. 1. Isolated rat kidneys were perfused at a constant pressure of 90 mmHg in a single-pass system with either a cell-free medium or a suspension of washed bovine red blood cells, free of the components of the renin-angiotensin system. In red blood cell perfused kidneys renal haemodynamics and sodium reabsorption corresponded closer to values observed in the intact rat than in cell-free perfused kidneys. 2. In red blood cell-perfused kidneys in the absence of plasma renin substrate autoregulation of renal blood flow was almost complete at pressures above 90 mmHg, provided that perfusion pressure was changed rapidly. 3. Renin release varied inversely with perfusion pressure within a pressure range from 50 to 150 mmHg; the greatest changes of renin release occurred, when perfusion pressure was reduced from 90 to 70 mmHg; maximal stimulation of renin release was observed at 50 mmHg. After reduction of perfusion pressure, renin release immediately started to rise and reached a new level within 5 min. Local reduction of perfusion pressure in small arteries and arterioles by the injection of microspheres induced a short-lasting decrease in renal plasma flow and a transient stimulation of renin release. 4. High concentrations of furosemide stimulated renin release by a direct intrarenal mechanism. 5. Isoproterenol stimulated renin release in low concentrations without a concomitant vasodilation, whereas high concentrations induced an increase in both renal plasma flow and renin release. The effects of isoproterenol were completely blocked by propranolol. 6. Sodium nitroprusside induced similar increases in renal plasma flow, as did high concentrations of isoproterenol, but only a small and slow increase in renin release was observed. 7. Angiotensin II (AII) suppressed renin release in concentrations corresponding to plasma levels measured in the intact rat independently of its vasoconstrictor effects, whereas vasopressin in antidiuretic concentrations did not affect renin release. 8. AII, AI, synthetic tetradecapeptide renin substrate (TDP), crude and purified rat plasma renin substrate induced a dose-dependent reduction in renal plasma flow. SQ 20 881, a competitive inhibitor of converting enzyme, and low doses of 1-Sar-8-Ala-AII (saralasin), a competitive antagonist of AII, did not change renal plasma flow, whereas high concentrations of saralasin had a vasoconstrictor effect on their own. 9. Saralasin inhibited the vasoconstrictor effects of AII and TDP to a similar degree. SQ 20 881 inhibited the vasoconstrictor effects of AI and purified renin substrate, but did not influence the actions of TDP and the crude renin substrate preparation. 10. From these data it is concluded, that AI is converted into AII within the kidney at a rate of 1-2%. The vasoconstriction induced by the crude renin substrate probably does not involve the AII receptors. TDP may act by itself on the AII receptors or via the direct intrarenal formation of AII...


Hospital-treated myocardial infarction and the general practitioner. Patients who had a myocardial infarction were interviewed with their spouses in hospital and after returning home. Advice about rehabilitation, such as that recently recommended by the Working Party of the Royal College of Physicians, was often not received. General practitioners have great opportunities for organising a graded programme of rehabilitation and may often communicate ideas more easily to their patients than hospital staff. However, some practitioners are reluctant to do this work.


Combination chemotherapy with adriamycin and cis-diamminedichloroplatinum in patients with neoplastic diseases. Forty-eight patients with a variety of advanced solid tumors were treated with a combination of adriamycin 50 mg/m2, and cis-diamminedichloroplatinum 50 mg/m2, every 2 to 4 weeks. Fifteen patients responded with a greater than 50% regression of measurable tumor; six with lung cancer; one, carcinoma of the breast; one, ovary; one, cervix; one, prostate; one, testis; one, maxillary sinus; and one, salivary gland, plus one patient with chemodectoma and one with adenocarcinoma of unknown primary. Responses lasted 1 to 18 months, with a median of 6 months. An additional six patients, including two with adenocarcinoma of the lung three with carcinoma of the cervix, and one with embryonal cell testicular carcinoma improved (25-50% regression of the tumor). Toxicity encountered included myelosuppression, azotemia, alopecia, nausea, vomitting, and stomatitis. Severe hematologic toxicity occurred only in those with compromised marrow function or with concurrent active hepatitis. Major potentiation of toxicity by the combination does not appear to have occurred.


[Planning work for the week for the purpose of more rational and effective use of available beds]. Planning the discharge and hospitalization of patients a week in advance, organization of checking up its fulfilment made it possible to improve the quality of investigation and treatment of patients, to settle the problems of hospitalization and to shorten preoperative stay of patients in the hospital. This was also promoted by the fact that patients entering the hospital underwent complete investigation in polyclinic conditions according to the patterns suggested by the hospital staff.


Clinical and dosimetric considerations in the radiation treatment of breast cancer. Preliminary results indicate that film dosimetry is a simple and precise method of determining volume dose distributions. Calculated dose distributions using computers or estimations of surface dose using TLD dosimeters are within 5% of the measured dose. Initial attempts to find suitable alternate techniques tailored to each clinical situation are encouraging.


A model of spinal cord injury.

A model of spinal cord injury. We report here a model of submaximal spinal cord injury, employing direct thoracic cord compression in ferrets. This yields consistent and reproducible long term consequences on local cellular structure and on locomotive function. We show also that both the anatomical and functional losses can be measured objectively using simple techniques, and that useful long-tract mediated locomotion is preserved after severe axonal losses at the site of injury.


Studies of nicotinic acetylcholine receptor protein from rat brain. Specific binding of 125I-labeled alpha-bungarotoxin to a 34800 X g pellet of a whole rat brain homogenate has been obtained at levels of 2 pmol toxin per g of whole brain with a Kd of 8-10(-9) M. Binding is reduced 90% by 10(-5) M (+)-tubocurarine chloride and 10(-4) M nicotine, whereas concentrations of 10(-4) M choline chloride, atropine sulfate and eserine sulfate have essentially no effect on toxin binding. These results compare closely with those obtained from binding studies with 125I-labeled alpha-bungarotoxin and soluble acetylcholine receptor protein preparations from Torpedo nobiliana; suggesting that this mammalian receptor protein is nicotinic in character. Extraction of the 34800 X g pellet with 1% Emulphogene yields a soluble fraction with specifically binds 125I-labeled alpha-bungarotoxin with a Kd of 5-10(-9) M. Nicotine and alpha-bungarotoxin at concentrations of 10(-5) M abolish toxin-receptor complex formation and carbachol and (+)-tubocurarine chloride reduce complex formation 35-40% at similar concentrations. Eserine sulfate, atropine sulfate, decamethonium, and pilocarpine had no effect on complex formation at concentrations of 10(-5) M.


Bilateral projection of the canine tooth pulp to bulbar trigeminal neurons. Unit activity was recorded extracellulary from neurons of the cat medulla following electrical stimulation of the ipsilateral and/or contralateral cannine tooth pulps. The majority of the cells (67%) were only responsive to ipsilateral stimulation. However, many (28%) responded to stimulation of either canine pulp and a few (5%) responsed to contralateral stimulation alone. The neurons were localized histologically in the necleus proprius of the rostral trigeminal nucleus caudalis (NVCaud) and in dorsal portions of the ventromedially contiguous lateral reticular formation (LRF). Cells exclusively responsive to ipsilateral stimuli had a relatively wide dorsoventral distribution. In contrast, 'bilateral' and 'contralateral' cells were situated only in the deep NVCaud-LRF border zone or in immediately adjacent portions of the LRF. Generally, ipsilateral stimuli evoked response bursts with shorter latencies, more spike potentials and briefer interspike intervals than equivalent contralateral stimuli. In experiments designed to study afferent interactions, a conditioning stimulus, applied to either the ipsilateral or the contralateral canine, preceded a test stimulus applied to the other canine at predetermined interstimulus intervals. Responses to the test stimulus were either totally or partially suppressed when intervals of moderate duration (90-500 msec) were used. However, responses to the test stimulus frequently were enhanced when the intervals were breif (less than or equal to 60 msec) or when the teeth were stimulated simultaneously. The results reveal that bilateral afferents from the pulps of the canine teeth converge upon neurons of bulbar trigeminal structures, that the neurons are differentially responsive to the activation of ipsilateral and contralateral pulpal receptors and that bilateral afferent barrages originating in the canine pulps interact to modulate the firing patterns of the neurons.


Genetic regulation of the antibody response to H-2Db alloantigens in mice. III. Inhibition of the IgG Response to noncongenic cells by preimmunization with congenic cells. When B10.A (5R) mice (H-12i5) are immunized with spleen cells from congenic B10 mice (H-12b), they respond to alloantigens of the H-2Db region by producing antibodies of only IgM type. In contrast, they produce both IgM and IgG antibodies when immunized with A.BY cells (H-2b) that carry other foreign cell surface antigens (non-H-2) in addition to H-2Db. Preimmunization of 5R mice with two injections of congenic cells leads to an H-2Db specific inhibition of the IgG response to a subsequent immunization with A.BY cells. It is concluded that congenic B10 cells fail to activate helper T cells which are necessary to induce the switch from IgM to IgG production. Instead T- or B-cell tolerance may be induced with prohibits the subsequent IgG response to A.BY cells, possibly by way of suppressor T cells which may act either on B cells directly or via helper T cells.


Fate in humans of the plasticizer, di-2-ethylhexyl phthalate, arising from transfusion of platelets stored in vinyl plastic bags. Platelet concentrates were shown to contain 18 to 38 mg/dl of a phthalate plasticizer (DEHP) which arose by migration from the vinyl plastic packs in which the platelets were prepared and stored. Transfusion of these platelets into six adult patients with leukemia resulted in peak blood plasma levels of DEHP ranging from 0.34 to 0.83 mg/dl (approximately 0.02 mg/dl plasma per mg DEHP administered per square meter of surface area). The blood levels fell monoexponentially with a mean rate of 2.83 per cent per minute and a half-life of 28.0 minutes. Urine was assayed by a method that would measure unchanged DEHP as well as all phthalic acid-containing metabolites. In two patients, at most 60 and 90 per cent of the infused dose, respectively, was excreted in the urine collected for 24 hours posttransfusion. These estimates, however, could be high due to the simultaneous excretion of DEHP remaining from previous transfusions or arising from uncontrolled environmental exposures.


Truncus arteriosus communis. Clinical, angiocardiographic, and pathologic findings in 100 patients. Salient clinical, hemodynamic, angiocardiographic, and pathologic findings are presented in 100 patients with truncus arteriosus communis, 79 of whom were studied at autopsy. In this study of typical truncus, all had a ventricular septal defect (type A). Truncus with a partially formed aorticopulmonary septum (type A1) was much the commonest form (50%). Cases with no remnant of aorticopulmonary septum (type A2) were second in frequency (21%). The distinction between types A1 and A2 could not be made with certainty in 9%, because these types merge into one another. Cases with absence of either pulmonary artery branch (type A3) were the least frequent form (8%). Truncus with interruption, atresia, preductal coarctation, or severe hypoplasia of the aortic arch (type A4) constituted 12%. The diagnosis of truncus is primarily angiocardiographic. The plane of the truncal valve in the lateral projection is distinctive. It tilts anteriorly, facing the patient's toes, which can be of assistance in differential diagnosis. The angiocardiographic features of type A4, although unfamiliar, are pathognomonic. Since the median age at death was only 5 weeks, and in view of the difficulties associated with pulmonary artery banding, our goal should be the surgical correction of truncus during the first and second months of life.


The immunologic identification of abnormal hemoglobins. Guinea pig antisera against human hemoglobin contain antibodies to multiple antigenic determinants. Hemoglobin variants having single amino acid substitutions affecting these determinants are recognized as antigenically deficient in a radioimmunoassay system. The multivalent antisera may be made more selective by appropriate absorption with hemoglobin variants. Three such antisera containing antibodies specific, respectively, for the beta-chain of Hb A, Hb S, and Hb C are described. They can differentiate among the hemoglobins of individuals who possess these variants alone or in any paired combination. Their discriminating ability is not hindered by the presence of 1,000-fold concentrations of Hb F and may therefore be extended to the early prenatal diagnosis of sickling disorders. The antisera described also detect several other variants of Hb A. The approximate location of the substitution in variants so detected can be determined by a complementation test. This is based on the finding that two antigenically deficient hemoglobins will complement each other in mixture provided their amino acid substitutions involve different determinants.


Evidence for dimensional independence in short-term memory of retarded individuals. This experiment was designed to assess the relative importance of dimensional similarity between test and interpolated problems and absolute difficulty of interpolated material as variables influencing short-term retention in the discrimination learning of retarded persons. Twenty moderate-to-borderline retarded individuals were taught a mixed series of two-trial discriminations consisting of: (a) relatively easy multidimensional ("junk") problems and (b) more difficult, form-relevant dotpattern discriminations. Test problems and interpolations were either dimensionally similar (both junk or both dot-pattern problems) or dissimilar (train one type of problem, interpolate the other). The major finding was that short-term forgetting occurred only when test and interpolated stimuli were dimensionally similar. Absolute difficulty of interpolations had no effect on retention. These results were interpreted as supporting Fisher and Zeaman's (1973) assumption of dimensional independence of memory processes in retarded persons. A secondary finding was that performance on dot-pattern discriminations varied with intelligence.


Abnormal or absent beta mRNA in betao Ferrara and gene deletion in delta beta thalassaemia. In patients with betao thalassaemia from Ferrara, beta globin mRNA sequences are either absent or structurally abnormal while in betao thalassaemia in Catania, beta globin mRNA sequences are present. In deltabeta thalassaemia there is a deletion of beta-like globin genes, while in betao Catania DNA, no beta globin gene deletion is detectable.


Effects of reserpine on tissue calcium and contractility of rat and rabbit aorta. Possible effects of reserpine on disposition and availability of tissue calcium, stores for excitation-contraction coupling in isolated rat and rabbit aortae were examined. Contral 40Ca uptake, 45Ca washout, and contraction in Ca2+-free medium (Ca2+-free PSS) indicate species differences in binding or disposition, apparent functional importance, and differential use of tissue calcium by adrenaline (Epi) and high K+. Rat aortae, normally refractory to Epi or high K+ after 7 min in Ca2+-free PSS, can gain labile calcium after brief exposure to Ca2+-rich PSS which supports short-lived responses to high K+ in Ca2+-free PSS. Rabbit aortae contain calcium stores which may sustain either Epi or high-K+ responses as well as more tightly held (or sequestered) stores released by Epi for contraction. After reserpine, decreased 45Ca uptake in a kinetically defined "fast" compartment likely to include membrane calcium could enhance availability of bound tissue as well as free Ca2+ in both species. Enhanced Epi response in Ca2+-free PSS is evidence of the former. Results suggest that increased availability of bound and possibly free calcium contribute to reserpine-induced supersensitivity, but supporting evidence will be required from tissue behavior after less rigorous treatment.


Effect of salicylate on the surgical inducement of joint degeneration in rabbit knees. Degenerative joint changes were produced in one knee of each of fourteen rabbits by surgical induction of instability. The involved knee in rabbits with and without systemic salicylate treatment was compared with the knee not operated on. Salicylate did not significantly change the activities of lysosomal enzymes in cartilage or synovial fluid, the uptake of tritiated thymidine, glycine, or 35S-inorganic sulphate by cartilage, or the histological manifestations of cartilage degeneration.


Mapping of the influenza virus genome. III. Identification of genes coding for nucleoprotein, membrane protein, and nonstructural protein. In previous communications we reported that the eight RNA segments of influenza A/PR/8/34 (HON1) virus could be distinguished from corresponding segments of influenza A/Hong Kong/8/68 (H3N2) virus by migration on polyacrylamide-urea gels. Examination of the RNA patterns of the two parent viruses and recombinants derived from them in concert with serological identification of surface proteins and analysis of the other proteins on sodium dodecyl sulfate gradient gels permitted the identification of the genes coding for hemagglutinin, neuraminidase, and the P1, P2, and P3 proteins (Palese and Schulman, 1976; P. Palese et al., Virology, in press). In the present report we have extended these observations using similar techniques to examine other recombinants and have identified the genes coding for the remaining virus-specific moving RNA segment as 1) and segment 6 of Hong Kong virus coding for the respective nucleoproteins, and that segment 7 of both viruses codes for the membtane protein and RNA segment 8 codes for the nonstructural protein. This completes the mapping of the influenza A virus genome.


Possible association between maternal indomethacin therapy and primary pulmonary hypertension of the newborn. Indomethacin purportedly arrests premature labor by inhibiting the production of prostaglandins. Animal experimentation has demonstrated that prostaglandins are involved in the neonatal rerouting of the circulation. Experience with two neonates with disorders circulatory adjustment at birth following prenatal exposure to indomethacin indicates that indomethacin may interfere with these adjustments in man and favor the development of the serious oxygen dependency known as primary pulmonary hypertension of the newborn.


Percutaneous trephine lung biopsy: evolving role. The Steele trephine was employed to perform 134 transthoracic biopsies in 120 patients. Resultant cores of tissue and "lung juice" provided a diagnosis in 89 percent (107) of the patients. Complications occurred in 41 percent (49) of the patients, the incidence being greatest in those with diffuse parenchymal disease. Most complications were of a minor nature. Although the precise role of trephine lung biopsy in the physician's diagnostic armamentarium remains to be defined, there is no doubt that it is a useful and safe technique in the elucidation of intrathoracic disease.


Major pulmonary resection for bronchogenic carcinoma in the elderly. The results of major pulmonary resection in 58 patients greater than 70 years of age were reviewed. The histological distribution and extent of nodal metastases in this age group are the same as in younger patients. The absolute five-year survival rate for the 55 patients undergoing curative resection was 30% (17 patients). It was 36% (11 patients) for those patients with squamous cell carcinoma and 22% (5 patients) for those with adenocarcinoma. The operative mortality was only 14% (8 patients). Of the 49 patients treated by lobectomy, 17 lived five years or more free of disease, whereas none of the 6 patients treated by pneumonectomy survived five years. The five-year survival rate of 30% in this series of elderly patients treated by major pulmonary resection makes resections in such patients with bronchogenic carcinoma worthwhile.


Psychiatric disturbances of aged patients in skilled nursing homes. The authors found that 85% of 74 patients supported by Medicaid in two skilled nursing facilities had significant psychiatric disorders in addition to serious multiple medical illnesses. Almost two-thirds of the psychiatric disturbances had not been diagnosed. Although staff were more concerned with the psychosocial than the physical aspects of patients' problems in more than half of the cases, they often had difficulty recognizing the legitimacy of psychological complaints and relating to patients with psychological disturbances. Staff were not clear about the orders for psychotropic medications that were prescribed for more than half of the patients. The authors point out that more psychiatric consultation is needed to ensure appropriate and effective care for psychiatric patients in such facilities.


Generalized urticaria to cyclophosphamide: Type I hypersensitivity to an immunosuppressive agent. A 6-yr-old white boy, Patient L. B., who was receiving cyclophosphamide (CY) for chemotherapy of a localized undifferentiated small cell sarcoma of the chest wall, developed generalized urticaria on three separate occasions immediately after CY administration. Serum samples were obtained 2 and 5 wk after the patient's last reaction and were examined for reaginic activity to the drug. Homocytotropic antibody activity to CY was found in the 2-wk postreaction sample by Prausnitz-Küstner type passive transfer skin testing to chimpanzees. This homocytotropic antibody was found to be of the IgE type, by means of an in vitro double-antibody coprecipitation assay measuring binding of free 14C-CY by specific immunoglobulin E and G fractions. Sera from 3 CY-treated patients who did not have adverse reactions to the drug and 6 normal, age-matched volunteers were used as control samples for the coprecipitation assay. None of the 3 CY-treated control patients had significant binding of 14C-CY by IgE when compared to normal individuals. However, it was found that all CY-treated patients had significant binding of 14C-CY by their IgG fraction when compared to normal subjects.


Rapid intravenous injection of tobramycin: suggested dosage schedule and concentrations in serum. A single, rapid intravenous injection of tobramycin in doses of 1.5 mg/kg or 80 mg administered to subjects with normal renal function produced a wide range of concentrations in serum. Serum concentrations peaked at greater than 20mug/ml in one third of the subjects, but these peaks were transitory. At 6 hr the serum levels were less than 2 mug/ml in all but one subject and were less than 0.1 mug/ml in 50% of the subjects. A dosage interval of 6 hr with a total dosage of 6 mg/kg per day is recommended. The variation in individual dose responses emphasizes the need for monitoring of levels of tobramycin in serum during treatment. Serum concentrations of 4-6 mug/ml 1 hr after injection can be considered indicative of adequate therapy. Levels in serum 6 hr after administration should also be monitored, particularly in the elderly. No immediate or delayed toxic effects were observed after intravenous injection of tobramycin.


Sensitivity of 15-day mouse fetal liver cells to erythropoietin (ESF). Its practical application to in vitro methods of ESF bioassay. Demonstration was made of equal reactivity of 13-, 14- and 15-day mouse fetal livers to graded concentrations of Erythropoietin "in vitro". In consequence, replacement by 15-day fetal liver of 13-day liver, currently used as material in Erythropoietin bioassays "in vitro", is proposed as a technical improvement on the method.


Normal values for ventilatory function in Rhodesian Africans. Forced vital capacity, forced expiratory volume in 1 sec and peak expiratory flow were measured in 141 African men and 102 African women aged 20 and above. Prediction formulae relating each measurement to height and age were drawn up and are presented. Rhodesian Africans have lower values for these measurements than subjects in some Europen and North American series, but broadly similar values to those of Africans elsewhere.


[Substitution of protein animal feed through lysine-supplemented high-protein wheat during the breeding and laying periods of hens. 5. Influence of graded lysine supplements fed during the laying period on the production characteristics of hens]. A large-scale trial was performed with 1500 laying hens which were kept under conditions of industrial egg production. The trial was carried out to investigate whether supplements of L-lysine would increase the food value of high-protein wheat and in which way a proportional decrease of dietary protein by 50% would affect the egg production of the hens. Additionally, studies were made to investigate whether the egg production of the hens was influenced by changes in the plane of feeding of the young hens during their rearing period. Both the young hens and the laying hens received isonitrogenous and isocaloric rations containing 4 graded levels of lysine. All of the 16 groups of the hens received a different diet. This was achieved by suitable combinations within the feeding regime. The control animals received, at all periods of age, food prepared on the basis of government-controlled standard rations. No significant differences in the level of food consumption were observed between the experimental groups whereas the control birds consumed more food (124 g per bird/day) than the experimental hens (120 g per bird/day; alpha less than 0.05). The levels of production were in all groups associated with the lysine content of the ration. The lysine-deficient birds produced the smallest eggs, layed the smallest number of eggs and consumed the largest quantity of food per 100 g of egg weight. The egg production of the hens in groups 11 and 12 (receiving a fish meal ration during the rearing period and a fish meal or wheat ration + 0.15% L-lysine during the laying period) was 70% of that of the birds in the control groups. The results of the present trial suggest that lysine-supplemented high-protein wheat rations for laying hens could be used but care should be taken that the plane of feeding for the young hens will not be too much restricted.