Naja melanoleuca (forest cobra) venom. The amino acid sequence of phospholipase A, fraction DE-III. Reduced and S-carboxymethylated phospholipase A (Fraction DE-III) from Naja melanoleuca venom was digested with trypsin, chymotrypsin and thermolysin. The resulting peptides were purified by ion-exchange chromatography on DEAE-cellulose, gel filtration on Sephadex G-25 or G-50 and chromatography and electrophoresis on paper. The amino acid sequences of the intact enzyme and the pur peptides were determined by the Edman procedure, either through the use of the automatic sequencer or by manual manipulation. The chymotryptic digest provided the necessary overlapping peptides which allowed the alignment of the tryptic peptides into a single chain of 119 amino acids. The amino acid sequence of N. melanoleuca phospholipase A shows a high degree of homology with phospholipases A from Bitis gabonica and also from porcine pancreas.


The amino acid sequence of phospholipase A, fractions DE-I and DE-II. The complete amino acid sequences of phospholipase A (Fractions DE-I and DE-II) from Naja melanoleuca (Forest cobra) have been elucidated. The reduced and S-carboxymethylated isoenzyme were digested with trypsin and thermolysin and the peptides were purified by ion-exchange chromatography, gel filtration and chromatography or electrophoresis on paper. The Edman procedure, either through the use of the automatic sequencer or by manual manipulation, was employed to obtain the sequences of the intact isoenzymes and the pure peptidesmthe thermolysin digest provided the necessary overlapping peptides which allowed the alignment of the tryptic peptides of Fraction DE-I. The tryptic peptides of Fraction DE-II were either identical or homologous to the tryptic peptides of Fraction I and Fraction III [12] and were aligned in the same order as that of Fractions DE-I or DE-III. The amino acid sequence of N. melanoleuca phospholipase A, Fraction I, shows a high degree of homology with Fraction DE-II and also with Fraction DE-III, previously reported on [12].


Platelet antiheparin activity. The isolation and characterisation of platelet factor 4 released from thrombin-aggregated washed human platelets and its dissociation into subunits and the isolation of membrane-bound antiheparin activity. Platelet factor 4 was isolated by gel filtration from the soluble release products of thrombin-aggregated washed human platelets as a proteoglycan-platelet factor 4 complex of molecular weight 358 000, Stokes radius (r-s) of 14.0 nm, sedimentation coefficient (s) of 7.1 S and frictional ratio (f/f-o) of 3.04. The complex was dissociated at high ionic strength (I equals 0.75) and the proteoglycan separated from platelet factor 4 by gel filtration. Platelet factor 4 had a molecular weight of 27 100, r-s of 2.52 nm, s of 2.4 S and f/f-o of 1.26, was insoluble under physiological conditions but readily soluble at pH 3. Under these conditions platelet factor 4 dissociated into four subunits with a molecular weight of 6900, r-s of 1.92 nm, s of 0.8 S, and f/f-o of 1.52. Qualitative N-terminal amino acid analysis showed the presence of glutamic acid or glutamine as the major end group. Platelet factor 4 was compared with protamine sulphate, which has similar biological properties, by electrophoresis at pH 2.2, in which both migrated as single bands but with differing mobility, and by amino acid analysis which showed a more normal distribution of residues than occurred in protamine sulphate. Of the basic amino acids platelet factor 4 (molecular weight 27 100) contained 5.97% arginine, 3.18% histidine, and 12.31% lysine compared to protamine sulphate with 64.2% arginine, 0.6% lysine and no histidine. A partial specific volume (v) of 0.747 was calculated for platelet factor 4 from its amino acid analysis. A membrane fraction with antiheparin activity, an isopycnic density of 1.090-1.110 and r-s of 15-35 nm, was also isolated by sucrose density gradient centrifugation from the ultrasonicated insoluble platelet residue remaining after thrombin-induced aggregation of washed human platelets. Trypsin treatment of the membrane fraction neither solubilised nor destroyed the activity.


Polynucleotide phosphorylase from Streptomyces aureofaciens: purification and properties. 1. Polynucleotide phosphorylase from a chlortetracycline-producing strain of Streptomyces aureofaciens was isolated by Polymin P fractionation. Using chromatography on DEAE-cellulose and Sephadex G-150 the enzyme, which appears homogeneous in gel chromatography and sedimentation analysis, was purified 2000-fole giving a final yield of 15%. 2. The sedimentation coefficient (s-o 20, w) of the native enzyme in 0.2 M NaCl is 9.15 S and its molecular weight is 210 000 plus or minus 15 000. Molecular weight estimated by sodium dodecylsulfate gel electrophoresis was about 100 000. 3. We have determined the optimal conditions for nucleoside 5'-diphosphate polymerization, their phosphate exchange and phosphorolysis of polyribonucleotides catalysed by polynucleotide phosphorylase from S. aureofaciens. 4. Chlortetracycline is a competitive inhibitor of S. aureofaciens polynucleotide phosphorylase. 5. Polynucleotide phosphorylase is activated in the polymerization reaction by ionic strength (K+, Na+, NH4+) while polyribonucleotide phosphorolysis is activated only by NH4+.


The two human chymotrypsinogens. Purification and characterization. The two chymotrypsinogens present in human pancreatic juice have been purified and characterized. The zymogens are two immunologically and electrophoretically different proteins. Chymotrypsinogen A, the major chymotryptic component (90% of the total potential N-acetyl-L-tyrosine ethylester activity) is stable in acidic medium. By its molecular weight (approx. 24 000), specific activity (530) and amino acid composition, human chymotrypsinogen A resembles chymotrypsinogens A and B form bovine and porcine pancreas. Chymotrypsinogen B is a minor chymotryptic component (7% of the total potential N-acetyl-L-tyrosine ethylester activity) unstable in acidic medium with a molecular weight slightly higher (approx. 27 000) and a specific activity slightly lower (300) than chymotrypsinogen A. The last 3% of the total potential N-acetyl-L-tyrosine ethylester activity corresponds to a proelastase that we have partially characterized.


Isolation of a glycoprotein and proteodermatan sulphate from bovine achilles tendon by affinity chromatography on concanavalin A-Sepharose. A fraction was isolated from a 3 M MgCl2 extract of bovine achilles tendon on the basis of its affinity for collagen. Affinity chromatography of this material on concanavalin A-Sepharose yielded a mixture which comprised a glycoprotein of approximate molecular weight 60 000 and two constituents containing hexuronic acid. The existence of a complex between the glycoprotein and material containing hexuronic acid was demonstrated by chromatography on Sephadex G-200 and by equilibrium sedimentation in CsCl density gradients. The complex was completely dissociated in 4 M guanidinium chloride. One of the constituents containing hexuronic acid was identified as a proteodermatan sulphate of low molecular weight and which had an abnormally high protein content (45-50%) and low buoyant density (1.46 g/ml) for a proteoglycan. The denser of the two molecules containing hexuronic acid appeared to be a normal proteoglycan, with a low protein content (11%). Analyses are given for the glycoprotein and the proteodermatan sulphate.


Carbohydrate binding specificity of the lectin from the pea (Pisum sativum). Hapten inhibition measurements on the precipitin reaction between Pisum sativum lectin and Pichia pinus phosphomannan showed the lectin to bind D-mannose, D-glucose, D-fructose and L-sorbose. Unmodified hydroxyl groups at the C-4 and the C-6 positions of the D-glucopyranose ring were essential for binding to the protein. Modification of the C-2 hydroxyl group was allowed in the D-glucopyranose ring but not in the D-mannopyranose configuration. Substitution of the hydroxyl hydrogen atom at the C-3 position of D-glucose increased the binding efficiency. With the exception of gentiobiose, the beta-linked glycobioses tested were not bound to the lectin, whereas the alpha-linked glycobioses were potent inhibitorsmin general, the P. sativum lectin was found to be less sensitive to structural variation of inhibiting carbohydrates than concanavalin A, the lectin from Canavalia ensiformis.


Improved purification of cathepsin B1 and cathepsin B2. An improved purification of the cathepsins B1 and B2 from bovine spleen is described. In addition to the formerly used procedure, chromatography with DEAE-Sephadex or -cellulose and mercurated agarose is used. Both enzymes are obtained in an electrophoretically pure form but consist of two or more isoenzymes. The isolation procedure leads to enzymes with high specific activities in satisfactory yields. Cathepsin B1 is frequently accompanied by small amounts of an arylamidase-like enzyme that hydrolyzes leucine p-nitroanilide. However, very probably, cathepsin B1 itself has a low activity toward this substrate too. Cathepsin B2 has a comparatively high activity with its characteristic though not specific substrate, alpha-N-benzoyl-L-arginineamide, whereas the activity toward haemoglobin is far lower. Both enzymes possess an essential SH group and require EDTA and a mercaptane for full activity, but their stability is markedly impaired by storage at higher thiol concentrations; Some other properties of the enzymes are also discussed.


Isolation, composition and functional properties of alpha-lactalbumins from several species. 1. Galactosyltranferases were isolated from bovine and pig milk and alpha-labtalbumins from milks of cow, pig, dog, rabbit, baboon and chimpanzee. 2. Alpha-Lactalbumins were characterized by gel electrophoresis, amino acid composition, ultraviolet absorbance, and ability to promote synthesis of lactose by the galactosyltransferases. 3. The order of K-m values of the various alpha-lactalbumins was the same with both cow and pig galactosyltransferases and differed from the order predicted from taxonomical classification of the species. 4. A280/A260 abosrbance ratios were correlated with K-m values suggesting involvement of tyrosyl groups in the function of alpha-lactalbumin.


Evaluation of the precision of difference chromatography. Difference chromatography, a method of chromatography in which elution volumes are compared directly, has been tested for precision under conditions used for estimating the relative dissociation of human oxyhaemoglobin and methaemoglobin and their hybrid. The test was carried out by first establishing a steady flow of haemoglobin through a column of gel particles, and then interrupting the flow temporarily by injecting a known weight of buffer solution, which thus simulated a small difference in elution volume. The dip in absorbance on the elution record was evaluated by digital methods. Statistical analysis of the results showed that the injected weight was estimated from the elution record with a standard error of plus or minus 0.02 g. Since the elution weight was about 50 g, the precision of difference chromatography must approach that of difference ultracentrifugation, and has obvious potential for the accurate comparison of the molecular weights of polypeptides in strongly dispersing media.


The relevance of the structure of lysine bound to Sepharose for the affinity of rabbit plasminogen;. The features of the structure of lysine, linked to Sepharose by the alpha-amino group, which are important for affinity chromatography of rabbit plasminogem were studied. Nine lysine and lysine-like conjugates, including epsilon-aminohexanoic acid DL-norleucine, DL-alpha-aminoadipic acid, DL-alpha-epsilon-diaminopimelic acid, cadaverine L-ornithine, L-arginine and D-lysine, were prepared; Using labelled rabbit plasminogen added to plasma, the ability of each conjugate to absorb plasminogen and separate the allomeric forms, type I and type II, during epsilon-aminohexanoic acid gradient elution was compared to Sepharose-L-lysine. Plasminogen had no affinity for Sepharose-epsilon-aminohexanoic acid, and was only weakly attracted by Sepharose-norleucine, Sepharose-cadaverine and others. Sepharose-ornithine held a greater attraction to the protein but the strongest binding was obtained with Sepharose-arginine. The affinity of plasminogen type I was always less than type II for the Sepharose-lysine analogues and the recovery of type II greater than type I from Sepharose-ornithine and Sepharose-arginine. Plasminogen affinity was in the order of Sepharose-arginine greater than Sepharose-lysine greater than Sepharose-ornithine. However, because of the present difficulty in recovering plasminogen from Sepharose-arginine the use of Sepharose-lysine in the affinity chromatography of rabbit plasminogen remains unchallenged. It is concluded that binding of rabbit plasminogen to conjugates of lysine and its analogues is determined by the presence of both a free carboxyl and a free amino group and that the distance between these groups is critical.


Vitamin D-dependent calcium-binding protein from rat kidney. A calcium-binding protein has been partially purified from rat kidney. It is found in the cortex, but not in the medulla. It is Vitamin D-dependent, as it occurs in normal, but not in Vitamin D-deficient rats. The molecular weight is 28 000, more than twice that of the Vitamin D-dependent calcium-binding proteins from rat intestinal mucosa. The apparent dissociation constant of the partially purified renal calcium-binding protein is approx. 10-5 M.


The protein composition of rat parotid saliva and secretory granules. Rat parotid saliva was collected by surgical cannulation of the ducts and stimulation with pilocarpine; The secreted salivary proteins were resolved on columns of DEAE-Sephadex into five major Fractions, I-V, which were characterized by polyacrylamide disc gel electrophoresis, amino acid analyses and enzymatic assay. Rat parotid secretory  granules were isolated by density gradient centrifugation and lysed in hypotonic buffers. Granule content proteins were resolved and examined by the same techniques as for secreted proteins. In both experiments, Fraction I contained RNAase and a major unidentified protein, M1, Fraction II contained the isoenzymes of amylase; DNAase was present in Fraction III and, to a lesser degree, in Fraction IV. The proportions of the enzyme-containing peaks were the same in saliva and granule contents. Fractions IV and V contain proteins of unknown function; Fraction IV contains exceptionally high levels of glutamic acid, glycine and proline in its protein moieties and approx. 6-8% neutral sugars.


The thermochemistry of reactions between alpha-s1-casein and calcium chloride. The enthalpies of reactions between alpha-s1-casein and Ca2+ in solution were measured using a gradient layer calorimeter. The reactions are exothermic between 0 and 4.3 mM CaCl2. In the region of 4.3 mM CaCl2 there is a change to an endothermic reaction corresponding to micellisation. A calcium-binding curve has been obtained under the same conditions as the calorimetry experiments and this shows two sigmoidal binding phases. Turbidity measurements show that there is an association process corresponding to the second sigmoidal phase. A tentative interpretation of the heat curve in the region before micellisation is given in terms of the site binding of Ca2+, conformational changes in the protein and association. The main thermal processes are taken to be exothermic intramolecular hydrogen bonding induced by calcium binding and endothermic hydrophobic bonding.


Metabolism of long-chain isoprenoid alcohols. Incorporation of phytol and dihydrophytol into the lipids of rat brain. [U-14-C]Phytol (3,7,11,15-tetramethylhexadec-2-en-1-ol) and [U-14-C]dihydrophytol (3,7,11,15-tetramethylhexadecanol) were administered intracerebrally to 18-day-old rats and incorporation of radioactivity into brain lipids was determined after 6 and 24 h. Radioactivity from [U-14-C]phytol was found in free phytenic (3,7,11,15-tetramethylhexadec-2-enoic), phytanic (3,7,11,15-tetramethylhexadecanoic) and pristanic (2,6,10,14-tetramethylpentadecanoic) acids, in phytanic and pristanic acid moieties of neutral and polar lipids, and in esters of phytol. In addition, evidence is presented for the utilization of phytol to form 1-O-phytenyl-2-acyl glycerophosphatides. Radioactivity from [U-14-C]dihydrophytol was found in free phytanic and pristanic acids, the corresponding acyl groups of neutral and polar lipids, esters of dihydrophytol and 1-O-phytanyl-2-acyl glycerophosphatides. Incorporation of either substrate into O-alkylglycerols was very low, and labeled branched-chain alk-1-enylglycerols could not be detected.


The activation of short-chain fatty acids by the soluble fraction of guinea-pig heart and liver mitochondria. The search for a distinct propionyl-CoA synthetase. 1. The ATP dependent acetyl-, propionyl- and butyryl-CoA synthetase activities were measured in the soluble fraction of both guinea-pig heart and liver mitochondria. 2. When measured in 300 mM Tris-HC1, the V of propionate activation in heart (equals 892 munits/mg protein) is much higher than the V of acetate (equals 637 munits/mg protein) and butyrate activation (equals 143 munits/mg protein. Fatty acid competition experiments, however, clearly show that most of the propionate activation (Km equals 7.94 mM) is caused by the acetyl-CoA synthetase (EC 6.2.1.1) (Km for acetate equals 0.8 mM), while the remaining activity is probably caused by a butyryl-CoA synthetase (Km for butyrate equals 0.83 mM). This indicates that in guinea-pig heart the presence of a distinct propionyl-CoA synthetase is very unlikely. 3. In liver a completely different pattern of short-chain fatty acid activation is found: low acetate activation and moderate propionate and butyrate activation. Substrate competition experiments and kinetics of fatty acid activation indicate that in this tissue a distinct propionyl-CoA synthetase is present with high affinity for propionate (Km equals 0.6 mM) and some affinity towards acetate and butyrate (Km values respectively 11 mM and 5.4 mM).


Colchicine-induced inhibition of plasma lipoprotein lipase release in the intact rat. The release of plasma lipoprotein lipase by heparin was studied in fed and food-deprived rats pretreated with colchicine and vinblastine. Four hours after the administration of either drug the lipoprotein lipase activity released by heparin was only half of that found in controls. Colchicine affected the release of both protamine-sensitive and protamine-resistant lipoprotein lipase. It is suggested that colchicine and vinblastine interfere with the transport of lipoprotein lipase from the site of its storage to the vascular cell surface.


The biosynthesis and turnover of different molecular species of rat testicular choline phosphoglycerides and triacylglycerols following intratesticular injection with (1(3)-14C)glycerol. Rats were sacrificed 1, 4, 8, 24, 48 and 96 h after an intratesticular injection with [1(3)-14-C]glycerol. The choline phosphoglycerides and triacylglycerols were isolated and the choline phosphoglycerides were converted to diacylglycerols by treatment with phospholipase C. These diacylglycerols and the triacylglycerols were fractionated into molecular species using argentation thin-layer chromatography. At early time periods the percent of radioactivity incorporated into the oleate- and linoleate-containing diacylglycerols exceeded the molar contribution of these two fractions. The percent of radioactivity in the linoleate fraction declined rapidly at later time periods while the percent of radioactivity in the oleate fraction also declined but not as rapidly. In contrast the percent of radioactivity in the arachidonate and docosa-4,7,10,13,16-pentaenoate-containing diacylglycerols at early time periods was lower than the molar amounts of these diacylglycerols. At later time periods the percent of radioactivity in these two fractions increased. These findings suggests that oleate- and linoleate-containing choline phosphoglycerides are made largely by total synthesis while a deacylation-acylation pathway plays a major role in the biosynthesis of the highly unsaturated choline phosphoglycerides. The calculated apparent half-lifes for the oleate-, linoleate-, arachidonate- and docosa- 4,7,10,13,16-pentaenoate-containing diacylglycerols were respectively 35, 18, 41 and 46 h. The percent incorporation of [1(3)-14-C]glycerol into the various triacylglycerol fractions did not differ markedly from the molar contribution of the respective molecular fractions suggesting that the various molecular species of rat testicular triacylglycerols are all synthesized and turn over at about the same rate.


The occurrence of dihydrophytyl wax esters in bovine rumen liquor. The purpose of this study was to examine the bovine rumen liquor wax esters and compare them with those of the dietary white clover leaf and stem tissues (Body, D.R. (1974) Phytochemistry 13, 1527-1530). The major fatty acid composition of the total bovine rumen liquor wax esters was made up of saturated fatty acids (62.1%) including palmitic (20.4%), stearic (16.2%) and other acids over the n-C19-n-C30 range (20.5%). However, 13.2% phytanic acid was also present. Of the two major fatty alcohol components, dihydrophytol (33.4%) and triacontanol (20.2%) were the principal contributors. Approximately 30% of the total intact rumen liquor wax esters was a series of dihydrophytyl wax esters. These represented 0.6% of the total rumen liquor lipid extract. Individually they were comprised of dihydrophytyl palmitate (0.05%), dihydrophytyl phytanate (0.10%) and dihydrophytyl n-C18 fatty acids (0.45%). The appearance of saturated isoprenoid moieties was the most striking difference between the composition of rumen liquor and dietary white clover wax esters.


[Action of microbial flora of the digestive tract on the metabolism of bile acids in the rat (author's transl)]. An isotopic balance is established in rats receiving a regular feed intake of [4-14-C]cholesterol so that various chemical species of bile acids have the same specific activity. This property is used to study bile acids distribution in the rat liver, digestive tract and fecal excretion. Bile acids are separated by thin-layer chromatography, radioactivity is determined by liquid scintillation, and the mass by 3-hydroxysteroid dehydrogenase action. The resulting comparative study made between the germ-free rat (axenic rat) and the rat exposed to microbes ("holoxenic" or conventional rat) receiving a semi-synthetetic feed, shows the influence excercised on the metabolism by the microbial flora of the digestive tract. This study confirms that the axenic rat compared to its holoxenic homologue has a higher bile acids pool and a lower fecal excretion. At all levels of the digestive tract (small intestine and the whole caecum and large intestine), probably as well as in the liver, the total amount of bile acids which is observed in the axenic rat is about twice the amount observed in the holoxenic rat, but fecal excretion is decreased by 20%. Values obtained by this method are higher than those previously observed by other authors using gas-liquid chromatography or [14-C]cholic acid isotopic dilution. This study also confirms that cholic and beta-muricholic acids are the main bile acids in the axenic rat and in addition establishes that in this animal bile acids composition is complex and varies from the small intestine to the feces. Besides cholic, alpha- and beta-muricholic, chenodeoxycholic and ursodeoxycholic acids, unidentified chemical species constitute 21% of the whole in the feces. Comparing the compositions observed in axenic and holoxenic rats in this experiment, it could not be determined if the relative activity of the two pathways of bile acid biosynthesis is deeply or only slightly changed by the presence of microbial flora. This is because of a large fraction of unknown composants in the feces of the axenic rat and the extreme complexity in the feces of the holoxenic rat.


Bile acids. XLIV, quantitation of bile acids from the bile fistula rat given (4-14C) cholesterol. The bile acids derived from [4-14-C]cholesterol administered intracardially to rats with cannulated bile ducts were identified and quantitated. Over a period of 28 days about 90% of the administered 14-C was found in bile of which 73% was retained in the biliary acid fraction. [7beta-3-H]cholic acid, alpha-muri[3beta-3-H]cholic acid, beta-muri[3beta-3-H]cholic acid and litho[3beta-3-H]cholic acid were prepared with specific activities of about 30 muCi/mg by reduction of appropriate ketonic precursors with NaB3H4 and were added to the biliary acid fraction. After separation and purification of the bile acids, cholic, chenodeoxycholic, alpha- and beta-muricholic acids accounted for 70, 16, 7.5 and 6.1%, respectively, of the 14-C in the biliary acid fraction. The specific activities of these isolated 14-C-labeled acids were almost identical. Lithocholic acid accounted for a maximum of 0.2% and ursodeoxycholic acid and 7-oxolithocholic acid could account for no more than 2% of the biliary 14-C. Gas-liquid chromatography on 3% OV-17 of the trimethylsilyl ether derivatives of the methyl esters of the common bile acids of rat bile results in their complete separation and provides a convenient means of estimating the relative proportions of these acids in rat bile. By this method, the relative amounts of the four major acids, cholic, chenodeoxycholic, alpha- and beta-muricholic acids were 63, 20, 8 and 6%, respectively.


Inhibition of cholesterol esterification by polyene antibiotics in blood plasma. The inhibition of the lecithin-cholesterol acyltransferase reaction in human plasma by the polyene antibiotics, levorine and amphotericin B, has been demonstrated by in vitro experiments. It is suggested that the inhibition of the esterification reaction is the result of interaction between polyenes and non-esterified cholesterol.


Specific effects of triarimol on sterol biosynthesis in Ustilago maydis. 1. Triarimol (2 mug/ml) severely inhibited ergosterol synthesis in sporidia of Ustilago maydis. In control cells ergosterol accounted for 70-85% of the total sterols, In sporidia treated 9.5 h with triarimol the total sterol content was not appreciably reduced; however, ergosterol constituted less than 4% of the sterol fraction. In treated cells 95% of this fraction was composed of 24-methylene-dihydrolanosterol, obtusifoliol and 14alpha-methyl-delta-8,24(28)-ergostadienol. These three sterols are proposed as the first intermediates of ergosterol biosynthesis in U. maydis. 2. This study indicates that in U. maydis triarimol inhibits three reactions in ergosterol biosynthesis, all of which involve the D ring and the side chain of the sterol molecule: demethylation at C-14; introduction of the C-22(23) double bond; and reduction of the C-24(28) double bond. The nature of inhibition suggests triarimol may bind to a sterol-sterol carrier protein complex and prevent interaction of the complex with enzymes necessary to carry out reactions in the affected region. 3. Cells treated with triarimol for 9.5 h contained approximately 100 times the quantity of free fatty acids found in control cells. There were qualitative differences in free fatty acids and fatty acids of the di- and triglycerides in control and treated cells.


Release of lipoprotein lipase from fat cells in vitro. Release of lipoprotein lipase from rat fat cells incubated at 20 degrees in medium with albumin, but without glucose proceeded at a constant rate for 30 min. The initial rate of release was increased when serum was present in the medium. Maximal stimulation (100-300%) was produced with 3.8% serum. The maximal increment in release caused by serum was always greater than that produced by heparin and when both were added release was greater than it was with either one alone. The active component(s) of serum, nondialyzable and stable for 30 min at 56 degrees C, was present in sera from humans and rats in the fed or fasted state. Glucose plus insulin (but neither alone) enhanced the rate of lipase release in the presence of serum but not in its absence. The half-life of the lipase in basal medium of 20 degrees C was 90 min. Heparin decreased this to about 50 min and serum markedly prolonged it whether or not heparin was present. Lipoprotein lipase activity in cells and fractions thereof was assayed in extracts of acetone powders. After centrifugation of fat cell homogenates at 600 times g for 15 min, only 50-60% of the activity was recovered in the supernatant. After centrifugation at 100 000 times g for 60 min, the supernatant contained about 10% of the total activity and the sediment 40%. In some experiments, most of the rest was recovered in the floating fat fraction. Total lipoprotein lipase activity of cells plus medium increased steadily during incubation of fat cells for 1h at 30 degrees C. The major increment occurred in the cells and activity in the medium was always less than 15% of the total. Our observations are consistent with the view that activation may be an important determinant of fat cell lipoprotein lipase activity as well as an integral part of the release process.


Monolayer characteristics of some glycolipids at the air-water interface. Surface pressure and surface potential versus molecular area data have been obtained for some galactosyldiglycerides and some galactosphingolipids at the air-water interface. The physical states of galactolipid monolayers (and bilayers) parallel those of the phospholipids. The molecular packing of the monolayers is determined primarily by the interactions between the hydrocarbon chains and chain melting causes the transition from condensed to expanded monolayer. Thus the long saturated chain cerebrosides from myelin have high chain-melting temperatures and form condensed monolayers with the chains in a quasi-crystalline array. The galactosyldiglycerides from chloroplast membranes contain polyunsaturate chains and form liquid-expanded monolayers. The surface potentials of monolayers of neutral galactosyldiglycerides are similar to those of equivalent lecithins; the contributions of the hydrated galactose and phosphorylcholine moieties to the surface potential are approximately equal. The various galactosphingolipid monolayers studied have quite different surface potentials; this indicates that relatively small variations in molecular structure which do not lead to appreciable changes in the average packing density can cause large changes in surface potential.


The interpretation of proton magnetic resonance linewidths for lecithin dispersions. Effect of particle size and chain packing. Two previously reported theoretical treatments of the effect of sonication on the PMR spectrum of phospholipid bilayer membranes have led to divergent conclusions regarding the effects of sonication on the structure of the bilayer membrane. In this report these two theoretical treatments will be critically reviewed, and it will be shown that only the theory of Seiter and Chan (Seiter, C.H.A. and Chan, S.I. (1973) J. Am. Chem. Soc. 95, 7541-7553) yields predictions which are in agreement with experiment. Analysis of available and newly acquired NMR results for sonicated bilayer vesicles of different sizes, both above and below the thermal phase transition, indicates that sonication does disrupt the regular molecular packing of the phospholipid molecules in these systems.


Modification of optical responses associated with the action potential of lobster giant axons. The sources of optical retardation changes and light scattering changes occurring during the action potential propagation of lobster giant axons have been investigated. A technique has been developed for resolving the total transmitted-light intensity change into a retardation change component, dI-r, and a forward direction light scattering change, dI-s. Trypsin, pronase, neuraminidase and hyaluronidase all reduce the magnitude of dI-r without diminishing the action potential, probably by cleaving charged saccharides. Dithiothreitol has no effect. This suggests that glycoproteins and hyaluronic acid polymers at the surface of the axon are involved in the optical responses, either by being passively realigned or by contributing to compression and expansion forces as the membrane electric field changes. Large dI-s responses are generated by trypsin and pronase treatment. The modifying effects of these proteases may be due to modification of the membrane or to increases in the refractive index of the medium surrounding the axon, since similar large dI-s, responses are produced by increasing the refractive index with sucrose. Since large reductions in dIr can be produced without concurrent reductions in the action potential, a significant portion of the optical retardation responses cannot be attributable to structural changes that are causally related to membrane ionic permeability changes during the action potential.


Some chemical characteristics of dimethylsuberimidate and its effect on sarcoplasmic reticulum vesicles. Sarcoplasmic reticulum vesicles treated with dimethylsuberimidate lose the capacity for ATP-promoted Ca2+ accumulation and show other properties indicative of leaky vesicles. As an aid to assessing whether this effect was caused by cross-linking or by hydrolysis products, characteristics of dimethylsuberimidate hydrolysis under incubation conditions used were measured. At pH 7.0, 25 DEGREES C, dimethylsuberimidate is hydrolyzed with an apparent first order rate constant of 0.016 min-1, to give dimethylsuberate as the principal product. The effect on ATP-promoted Ca2+ accululation was shown to be caused by partially and fully hydrolyzed products of the diimido ester, and not to cross-linking of membrane components.


Barrier properties of lecithin/lysolecithin mixtures. Light scattering, birefringence and X-ray studies showed that liposomes, with lipid molecules orientated in bilayers, are formed from egg licithin/lysolecithin mixtures up to 50 mol percent of lysolecithin; above this concentration much smaller mixed micelles are formed. Permeability studies demonstrated a dramatic increase in the permeability of the liposomes when the lyso concentration exceeds 22.5 mol percent. X-ray studies indicated a significant decrease in bilayer thickness with increasing lysolecithin concentration. It is suggested that decreased interaction energy between the lipid molecules in the bilayer is responsible for the inability of the thin bilayers to act as an effective permeability barrier.


Kinetics of macrotetrolide-induced ion transport across lipid bilayer membranes. Ion transport across lipid bilayer membranes in the presence of macrotetrolide antibiotics has been studied by stationary conductance and nonstationary relaxation methods. The results are discussed on the basis of a carrier model which has already been successfully applied to valinomycin induced ion transport. Again a kinetic analysis has been performed from which the single rate constants of the carrier model could be derived. In addition the equilibrium constant of complex formation in the aqueous phase could be determined. Measurements have been made for 4 macrotetrolides, for several ions and for various chain lengths of the lipid molecules composing the membrane.


The effects of the membrane-penetrating polypeptide segment of the human erythrocyte MN-glycoprotein on the permeability of model lipid membranes. The insoluble peptide, T(is), prepared by trypsin hydrolysis of the MN-glycoprotein (glycophorin) of the human erythrocyte has been incorporated into phospholipid membranes in the form of liposomes and black lipid membranes. The permeability of liposome membranes to 42K+ and of black lipid membranes to water and ions is increased significantly by the presence of the T(is) peptide. Electrophoresis measurements indicate that these effects are not due to the T(is) peptide carrying a net charge. The results suggest that the peptide causes local disordering of the bilayer membrane structures. This is considered in the light of findings published elsewhere: that the MN-glycoprotein penetrates through the cell membrane via a non-polar segment of its polypeptide chain, which is contained intact within the T(is) peptide; that the T(is) peptide is partially helical when associated with phospholipid and forms multimeric 8.0 nm structures within the hydrophobic plane of phospholipid bilayers.


A laser Raman spectroscopic investigation of phospholipid and protein configurations in hemoglobin-free erythrocyte ghosts. Configurations of both the protein and lipid components of human red blood cell ghosts are examined by laser Raman spectroscopy. Protein configuration is estimated from bands observed in the Amide III region at 1240-1267 cm-1 in water and the Amide I' region at 1630-1670 cm-1 observed in 2H2O. The protein fraction appears to contain 40-55 percent alpha-helix with little beta-configuration. The hydrophobic side chains of the phospholipid component, as interpreted from the 1060-1130 cm-1 C-C stretching region, are estimated to contain 55-65 percent all-trans rigid configuration. These estimated are within the limits set by other physical techniques.


Postreplication DNA repair in ultraviolet-irradiated Micrococcus luteus. Postreplication DNA repair was studies in three strains of Micrococcus luteus having different sensitivity to ultraviolet light: a wild type ATCC 5698, a ultraviolet-sensitive mutant G7, deficient in the incision step of repair and in ultraviolet-resistant transformant obtained from G7 by treatment with DNA of wild type cells, Trf(G7). It is shown that the G7 mutant has a low capacity for repair of postreplication DNA gaps compared with the wild type or Trf(G7). It seems to be that postreplication repari capacity contributes significantly to the ultraviolet resistance of M. luteus in addition to the excision repair. In contrast with G7 the size of the DNA fragments synthesized immediately after ultraviolet irradiation in the wild type (and Trf(G7)) seems to be much higher than that expected if each dimer produces one DNA gap in the daughter strand. Since this cannot only be explained by the excision of dimers from parental DNA we have suggested that a rapid repair of postreplication DNA gap occurs in M. luteus.


Early estrogen action. Stimulation of the synthesis of methylated ribosomal and transfer RNAs. The effects of estrogen on the rates of incorporation in vivo of radioactive uridine and Me-methionine, administered together, into RNA in the uterus of the ovariectomized adult rat have been measured. The ratio of incorporation of methionine to uridine during a 45-min labeling period was increased several-fold by hormone treatment. The increased rate of methylation was apparent in the uterus taken from the rat administered estrogen for 1 h, and the effect was more striking following 2 and 3 h of hormone treatment. This stimulation of methylation of RNA occurred in association with an increase in the whole-organ concentration of RNA. Analysis of the doubly-labeled uterine RNA on sucrose gradients revealed that the methylated species were mainly ribosomal and transfer RNA. These results show that very little methylation of RNA occurs in the atrophied uterus of the ovariectomized rat. During the first 3 h following estrogen administration to the ovariectomized animal, an increasing percentage of the newly synthesized RNA formed by the uterus is methylated ribosomal and transfer RNA. This result is discussed in light of recent studies of the efficiency of processing of ribosomal precursor RNA, as well as the synthesis of high-molecular-weight heterogeneous RNA in the early action of estrogen.


DNA-dependent RNA polymerases from Acanthamoeba castellanii: properties and levels of activity during encystment. Three DNA-dependent RNA polymerases have been isolated and partially purified from trophozoites of Acanthamoeba castellanii. Separated by DEAE-Sephadex chromatography, they have been designated polymerases, I, IIa and IIB according to their alpha-amanitin sensitivity and kinetic properties. I is completely insensitive to alpha-amanitin. IIa and IIb are sensitive to low concentrations (0.1 mug/ml) of alpha-amanitin; however, in order to achieve 100% inhibition much higher concentrations (130 mug/ml) are needed. Both I and II (a or b) have rather broad ionic strength optima (0.06--0.10 M (NH4)2SO4). All three prefer denatured over native DNA (I, 4:1; II, 2:1). Polymerase I utilizes magnesium better than manganese as divalent cation whereas II prefers manganese. When Acanthamoeba is transferred to a medium lacking nutrients, the cells undergo a synchronous differentiation resulting in cyst formation. In general agreement with the decrease in the rate of synthesis of its product (rRNA), the amount of polymerase I decreases relative to the amanitin sensitive polymerase(s). However, the absolute amount of polymerase I does not change. Rather, the levels of the amanitin sensitive enzymes increase during the first 10 h of encystment. Since the overall RNA synthesis rate decreases, these results suggest that the transcription rate is not controlled by specific enzyme levels alone.


Comparison of ribosomal proteins from neoplastic and non-neoplastic cells. Resolution by two-dimensional gel electrophoresis. Functionally active mammalian membrane-free ribosomes were prepared from rat forebrain, a solid tumor (mouse neuroblastoma C-1300) and a human tumor cell-line in suspension culture (HeLa). Proteins were extracted from these ribosomes and were resolved by a newly developed. Two dimensional gel electrophoretic procedure. Although the ribosomal protein patterns from the three preparations were generally similar, there were four obvious differences between the forebrain ribosomes and the ribosomes from the two neoplastic cell types.


In vitro stimulation of apoferritin synthesis by iron. The apparent induction of apoferritin synthesis by iron has been examined in cell-free systems from rat and rabbit liver. Both systems allowed the complete synthesis de novo of apoferritin isolated by chromatographic or immunological means. Addition of iron at levels of 0.2--1 mM specifically stimulated incorporation of radioactive amino acids into apoferritin purified after classical heat extraction. The effect was also observed when iron was added at the end of the incubation period in the absence of continuing protein synthesis. Further, iron addition had no effect on the amount of newly synthesised apoferritin subunits as estimated by direct immunological precipitation from the reaction mixture. These results suggest that iron acts at some stage subsequent to translation in stimulating apoferritin biosynthesis.


Light scattering from nucleated biological cells. The light scattered from nucleated biological cells has been investigated by using four different theoretical models: an opaque disk, a homogeneous sphere, an opaque ring, and a coated sphere. By comparing these four models, diffraction at the edges of the cell and the nucleus has been found to be the predominate scattering mechanism for nucleated biological cells at low angles. The scattering patterns of nucleated cells are found to have a fine lobe (high-frequency) structure dependent on whole cell size, and an envelope lobe (low-frequency) structure dependent on relative nucleus size. The models indicate that the present technique for measuring cell size with a single low-angle light detector is highly dependent on the nucleus to cell diameter ratio. Whole cell size is better estimated by the ratio of the outputs from two low-angle detectors.


Geometry of the human erythrocyte. I. Effect of albumin on cell geometry. The effects of albumin on the geometry of human erythrocytes have been studied. Individual red cells, hanging on edge from coverslips were photographed. Enlarged cell profiles were digitized using a Gradicon digitizer (Instronics Ltd., Stittsville, Ontario). Geometric parameters including diameter, area, volume, minimum cylindrical diameter, sphericity index, swelling index, maximum and minimum cell thickness, were calculated for each cell using a CDC 6400 computer. Maximum effect of human serum albumin was reached at about 1 g/liter. Studies of cell populations showed decreases in mean cell diameter of up to 6%, area 6%, and volume 15%, varying from sample to sample. The thickness of the rim was increased while that at the dimple was decreased. Studies of single cells showed that area and volume changes do not occur equally in all cells. Cells with lower sphericity indices showed larger effects. In the presence of albumin, up to 50% of the cells assumed cup-shapes (stomatocytes). These cells had smaller volumes but the same area as biconcave cells. Mechanical agitation could reversibly induce biconcave cells to assume cup shapes without area or volume changes. Experiments with de-fatted human albumins showed that the presence of bound fatty acids in varying concentrations does not alter the observed effects. Bovine serum albumin has similar effects on human erythrocytes as human serum albumin.


Analysis of fluorescence decay curves by means of the Laplace transformation. A computational procedure is described for the analysis of fluorescence decay data convolved with a lamp flash of finite width. The computer program calculates the ratio of the Laplace transforms of the decay and the lamp flash for different values of s to give the transforms of the impulse response for each value of s. These are set equal to the analytical Laplace transforms of the decay law involved. Solution of the nonlinear simultaneous equations yields the desired decay parameters. The method can be modified to analyze data that contains a component due to scattered light and can also provide essential information regarding transit time changes of the photomultiplier with changes in emission wavelength. The method was tested by the analysis of real and simulated data. The accuracy of the analysis depends on the degree of correlation among the parameters.


[Influence of ribose on 2,3-diphosphoglycerate concentrations in human erythrocytes]. The effect of ribose on the 2,3-diphosphoglycerate concentration (2,3-DPG) of human red blood cells was investigated and compared to the effect of inosine. Ribose as well as inosine increases 2,3-DPG. Most impressing effects can be demonstrated on stored blood, which is poor in 2,3-DPG concentration. Physiologic 2,3-DPG-concentrations, however, may be increased too. Generally the effect of inosine is greater than the effect of ribose. When ribose is applicated intravenously a slight increase in 2,3-DPG is found. Both compounds own characteristic side-effects, which diminish their value for therapeutical purposes.


[Autoradiographic demonstration of human basophils]. A new technique is described which enables a selective tinctorial visualization of human blood basophils in autoradiographic preparations. Consecutive administration of 5-aminoacridine hydrochloride and aldehyde fuchsin rendered a sufficient preservation of basophils preventing leaching of their water soluble granules. Extensive control tests provided sufficient evidence to exclude chemographic artifacts produced by other histochemical precedures. The technique eliminates the obstacles to study the kinetics of human blood basophils and their realtionshp to atopic hypersensitivity reactions applying tritiated thymidine and radioiodine labeled IgE or corresponding antisera.


[99mTc-sulfur-rhenium colloid and 111In-indium citrate in bone-marrow scintigraphy]. Bone marrow scinitigraphy using 111In-Indium-citrate and 99mTc-sulfur rhenium colloid was compared with each other in 6 male and 3 female patients. Our results in all patients were in favour of 111In-citrate which caused a better delineation of morphological details than 99mTc-sulfur rhenium colloid did. In the first days after intravenous administration of 111In-citrate, radioindium accumulated markedly in the pudendal region, this finding being more distinct in male patients than in female ones. On account of these results a considerable radiation dose has to be assumed in bone marrow scinitigraphy using 111In-citrate as long as the absorbed dose from 111In has not been estimated under consideration of the radioindium accumulation in the pudendal region. 111In-citrate should be applied in bone marrow scanning only exceptionally and 99mTc-sulfur rhenium colloid be preferred in the routine diagnostics of bone marrow.


Errors in decoding tone of voice during dyadic interaction. Predictions derived from evidence on projection, behavioural assimilation and attribution theory suggest differing ways in which one individual may perceive another's tone of voice in dyadic interaction. Pairs of male subjects undertook a competitive or cooperative counting task. One member of each pair was instructed that he could best influence the dyad's performance by the use of an angry or pleased tone of voice. The results indicate that while subjects in the cooperative condition tended to attribute their own tone of voice to their (neutral) partner, competitive subjects attributed a discrepant or neutral tone. The findings are interpreted in terms of attribution theory. Implications for studies of non-verbal communication are discussed.


The role of gaze in impression formation. Three hypotheses about the effects of amount of gaze on the impression created of six confederates were tested, using rating scales and free descriptions. The first hypothesis--that favourableness of impression would be a linear function of amount of gaze--was partly confirmed. The second hypothesis--that favourableness of impression would be a curvilinear function of amount of gaze--was not confirmed, nor was the third hypothesis--that effect of gaze would be a complex function of sex of subject and sex of gazer. The effects of gaze were not as marked as had been predicted. The free descriptions also provided data on the number and type of subjects who commented on the amount of gaze of the confederate. Three possible explanations of the results are discussed.


Encoding and decoding of ingratiation in modes of smiling and gaze. The present research is a first attempt to explore the ingratiation ingredients of non-verbal attractiveness. In particular, the ingratiator's paradox is investigated. This occurs when an increase in attractiveness of A makes B aware of an ingratiation attempt. A first experiment was set up to detect the ways in which ingratiation overtures are displayed in smiling and gaze during task-orientated behaviour. As predicted from empirical research and from Argyle & Dean's (1965) 'intimacy equilibrium' model, a higher percentage of smiling and gaze, together with more frequent smiling and longer gaze, occurred under the ingratiation than control condition. A second experiment investigated if the ingratiation and spontaneity behaviours that were displayed would be differentially evaluated. To this purpose, observers naive to the conditions were confronted with the videotaped behaviours. As predicted from ingratiation and attributional frameworks, ingratiation behaviours were more favourably evaluated, though they were considered as strategic in nature. This discussion highlights the subtlety involved in non-verbal ingratiation.


Covariation of conflict and mood in depression. The proportion of conflict involved in the personal problems of neurotic and depressed inpatients was assessed and correlated with the patients' mood over a period of several months. Mood correlated highly with conflict, i.e. when conflict was high, patients felt more depressed, fatigued and tense. Conflicts in the patients' beliefs about themselves also tended to correlate highly with mood, while conflict connected with their attitudes concerning their problems did not. Individual patients seemed to "react" to conflict with different kinds of mood.


Recall of medical advice: comprehensibility and specificity. Two laboratory experiments are described in which volunteer subjects read advice on how to develop self-control over eating behaviour. Formulating the communications in terms of specific instructions rather than general rules and the use of simple language as determined by a readability formula led to increased recall of the advice. These findings were confirmed in a naturalistic setting in which the same material was presented orally to patients receiving treatment for obesity. The results of these investigations are believed to have implications for improving communications between doctors and their patients and possibly for increasing the degree to which patients comply with medical recommendations.


A proposed basis for delusion formation within an information-processing model of paranoid development. A developmental model for paranoid behaviour is proposed within a broader theory of schizophrenic development. The model stipulates that given exposure to sustained aversive maternal control and a maternal communication style which is subtle and devious, the child comes to adapt with approach, stratagem-based behaviours and heightened vigilance for evaluative information (i.e. open adaptive style). The model, already supported empirically at several points, postulates that the delusion serves to organize an overextended and disorganized information-processing system for the person and generates its reinforcement from reduction of the anxiety associated with thought disorganization. The present study tested an aspect of this postulate. It was predicted that late-adolescent males who had an open style of adapting to experienced aversive maternal control would find the organization of disorganized evaluative cues uniquely reinforcing. Subjects were presented with an anagrams task made up of scrambled words which they were told came from a poll of mothers asked to evaluate their college-age sons. This task was followed by the individual tachistoscopic and slightly unfocused presentation of a longer series of words, again described as maternal evaluative terms. The longer series comprised the words used in the anagrams task and an equal number which were not. Reinforcement value of proper ordering of disorganized cues was inferred from the subject's heightened ability to detect previously solved anagram terms, indicating stronger learning of these terms upon prior solution. Males identified as open-style adapters were far more cognizant of words which they had previously ordered than other child-rearing groups, as predicted.


Impaired recall and the memory disturbance in presenile dementia. Although subjects with presenile dementia have a very poor ability to retrieve material previously presented for learning under conditions of free recall and the usual type of recognition test, their performance was found to be not significantly different from normal controls when the initial letters of the correct words were provided at the time of recall. This finding raises the possibility that the long-term memory failure in presenile dementia may not be due to an inability to establish new material in the long-term store, as had previously been thought, but to a difficulty in retrieving that information.


A two-stage model of attention in schizophrenia research. This paper attempts to indicate the relevance of Broadbent's recent views on the methods by which the systematic selection of information takes place, for the study of attention deficits in schizophrenia. A two-stage model of attention is briefly described, the distinction between "filtering" (stimulus set) and "pigeon-holing" (response set) being emphasized. In an attempt to clarify the nature of the schizophrenics attention defect, various experiments are reviewed and interpreted within Broadbent's theoretical framework. Studies indicate inefficiencies in both the "filtering" and "pigeon-holing" mechanisms. The need for experiments specifically designed to identify the locus of the schizophrenic attention defect, in various subgroups of schizophrenics, is indicated.


Plasma catecholamines in severely injured patients: a prospective study on 45 patients with multiple injuries. Plasma catecholamine levels were studied in 45 severely injured patients for 8 days after the trauma. Sixteen of the patients were classified as critically injured and 29 as seriously injured. The total plasma catecholamine values of the whole group immediately after the injury were almost twice as high as the eighth day reference values and remained significantly higher than these values for 6 hours after the trauma. On admission both the plasma adrenaline and noradrenaline levels were elevated. The plasma adrenaline levels on admission correlated with the blood volume replacement which was required within the first 6 hours. The plasma noradrenaline levels in the critically injured group were significantly higher throughout the observation period than in the seriously injured group. A corresponding difference was observed in the plasma adrenaline concentrations only during the first 12 hours. The results showed that strong stimulation of the sympathetic nervous system occurs in severely injured patients. Factors stimulating the sympathetic nervous system apparently included hypovolaemia, tissue hypoxia, acidosis and the pain produced by the trauma and therapeutic measures.


Postoperative problems and management after hepatic resection for blunt injury to the liver. The postoperative problems and management of 8 patients following right hepatic lobectomy for blunt liver injury are discussed. Multiple injury and in particular chest injury are of importance. Respiratory distress may be insidious in onset and must be anticipated. Most patients require at least temporary positive pressure ventilation. Liver regeneration is rapid and histological evidence of regenerative hyperplasia is present within 3 days of injury. Hypoglycaemia of a degree sufficient to threaten life may occur postoperatively, particularly in the first 48 hours. Jaundice may be a worrying postoperative feature, but the use of T-tube drainage after hepatic lobectomy will enable any possible extrahepatic obstruction to be excluded. Hypoproteinaemia and hypo-albuminaemia occur in the immediate postoperative period, and recovery rapidly follows the peak period of regenerative activity of the liver. Normal levels are regained by the fourth to sixth postoperative weeks. Prophylactic antibiotics have no place in the postoperative management of blunt liver injury. Haemorrhagic diathesis is common after hepatic resection and is of a complex nature. Intravascular coagulation may occur and factor V deficiency is common. The mainstay of treatment is transfusion of fresh blood.


The application of the 99Tcm phosphate bone scan to the study of breast cancer. Comparison of the gamma camera bone scan and radiological survey in 190 patients with breast cancer has confirmed the superiority of the bone scan in the positive diagnosis of skeletal metastases. In 47 patients with known (X-ray positive) bony metastases the scan was positive in all but 2, and in 24 (50 per cent) the scan showed more lesions than the X-ray had indicated. In 60 patients with a clinical suspicion of bone metastases but negative X-rays, the scan was positive in 29 (48 per cent). Eighty-three patients with primary breast cancer were studied at the time of initial presentation. All had a normal radiographic skeletal survey but 24 (27 per cent) had scan evidence of occult metastases. It is concluded that a bone scan is indicated in all patients with breast cancer where skeletal metastases are suspected. Repeated bone scans may introduce new therapeutic possibilities, by providing an earlier objective index of progression of metastatic disease and also by providing a quantitative method of assessing the response to therapy.


Cystic thyroid nodules.

Cystic thyroid nodules. Among 300 patients with thyroid nodules subjected to a needle drill biopsy, 62 were found to contain cystic lesions. Analysis of these 62 showed that in 17 the swelling had completely disappeared after the biopsy. The remaining 45 had residual or recurrent swellings. Operative treatment was carried out in 35, which consisted of 28 (80 per cent) nodular goitres, 6 (17-1 per cent) adenomas and 1 (2-9 per cent) carcinoma.


Motility changes in the antrum after proximal gastric vagotomy. The normal pattern of resting and post-prandial motor activity in the gastric antrum has been established by observations in 6 dogs. There was a gradual increase in the amplitude of contraction during the first 2 hours after eating; this was maintained for 3 hours and then declined. For the first 45 minutes terminal antral contraction occurred, partially retaining and triturating the gastric contents. After 45 minutes the waves became sequential, symmetrical, increased in vigour and actively pumped food into the duodenum. Vagotomy modified the mechanism of the antrum in various ways. Truncal and selective vagotomy reduced the work capability to 20 per cent of its normal value when recorded 1 month after operation. In both groups the waves were disorganized. Proximal gastric vagotomy abolished the braking mechanism and removed the initial inhibitory stimuli to antral motility. Within 1 month of operation the antrum had regained 58 per cent of its normal work capability and the contractions were well organized.


The management of bleeding acute gastric erosions: the role of gastric hypothermia. Eighty-eight consecutive patients with bleeding acute gastric erosions were reviewed with the aid of a computer. Of the eroisons, 31-8 per cent were related to the ingestion of ulcerogenic drugs and 40-9 per cent were associated with surgery and sepsis. The overall mortality was 20-5 per cent. Endoscopy was the most effective diagnostic examination. Conservative management was effective in the treatment of bleeding erosions due to the ingestion of ulcerogenic drugs. Gastric cooling successfully arrested haemorrhage from bleeding erosions due to surgery and sepsis.


Flatulent dyspepsia in patients with gallstones undergoing cholecystectomy. The incidence of flatulent dyspepsia and its relationship to gallbladder function has been studied in 100 consecutive patients with gallstones undergoing cholecystectomy. Thirty-three per cent of patients suffered significant flatulent dyspepsia of whom 80 per cent were cured or improved by operation. In 15 patients gastric function was studied pre- and postoperatively and it was noted that there was no difference in gastric emptying times between patients with flatulent dyspepsia who were cured by operation and those who remained symptomatic. Bacteriological studies on gallbladder bile from 39 patients suggested that infection within the gallbladder may be a factor in the causation of flatulent dyspepsia.


Traumatic haemobilia.

Traumatic haemobilia. A case of traumatic haemobilia which was diagnosed preoperatively, is reported. The site of bleeding was demonstrated by operative angiography through the right hepatic artery. The patient was treated by ligation of the common hepatic artery. Re-bleeding required a second operation and ligation of the right hepatic branch without adverse consequences on the hepatic function.


Rupture of subcapsular haematoma of the liver in a case of eclampsia. A case of rupture of subcapsular haematoma of the liver in a patient suffering from eclampsia is reported in which the patient survived. Peritoneal tap provided important diagnostic information. Surgical intervention with control of the liver haemorrhage provides the only chance of survival.


Temporary transcaecal ileal diversion in prevention of anastomotic leak in planned colonic surgery (an alternative to proximal colostomy). An alternative to proximal colostomy in planned colonic resection is reported in 30 patients. The physiopathological background for this procedure of temporary transcaecal ileal diversion is explained together with details of the operation and the results obtained.


A colostomy/ileostomy construction clamp. An atraumatic handleless clamp is described with which it has been found simpler to construct a terminal ileostomy or colostomy than with other forms of clamp in general use.


Ureteric obstruction due to sigmoid diverticulitis. Two cases in which the ureter was obstructed by an inflammatory diverticular mass are presented. The difficulties of distinguishing such a mass from a carcinoma both clinically and at operation are emphasized. In order to minimize the risk of ureteric injury and to facilitate planning of the operative procedure, it is recommended that intravenous pyelography should be performed before resection of a suspected diverticular mass.


The striatonigral degenerations. Putaminal pigments and nosology. Three new cases of striato-nigral degeneration (SND) are presented with particular reference to the identification of putaminal pigments. These were studied by histochemical methods, electron microscopy and elemental analysis. Three interrelated parenchymal perikaryal pigments were identified in the putaminal lesions of all 3 patients: a "haematin" pigment, neuromelanin and lipofuscin. The presence of neuromelanin in the putamen may be due to accumulation of dopamine in the synaptic terminals of the nigro-striatal pathway and its polymerization into pigment. This suggests that the putaminal atrophy is the primary lesion in SND which is a true supranigral form of parkinsonism. Clinically this manifests itself in predominance of rigidity over other parkinsonian symptoms and in a poor, or absent, response to treatment with L-dopa and anticholinergic drugs.


Electrophysiological correlates of pathology and surgical results in temporal lobe epilepsy. Routine pre-operative EEG studies as well as direct brain recording and stimulation carried out during operations were analysed for 59 patients subjected to a standard unilateral anterior temporal lobectomy for the treatment of epilepsy. All patients in the present series were 16 years old or older at the time of operation, which was invariably carried out under local scalp analgesia only. Electrophysiological findings was correlated with pathological changes noted in the resected temporal lobes, and with the effects of surgery upon seizure activity. Pre-operative EEG data correlated with each of four pathological categories when sphenoidal electrodes and intravenous barbiturate narcosis were emplyed. Thirty of 31 patients with mesial temporal sclerosis demonstrated medial temporal primary spike foci, frequently with independent contralateral and extratemporal secondary foci. In addition, one-third of these patients demonstrated unilateral focal decreased barbiturate-induced fast activity in the corresponding sphenoidal to ear channels. Twelve patients with other specific medial focal lesions (mostly hamartomas) also had medial temporal primary foci, often with independent contralateral secondaries but never with extratemporal foci. Two patients in this group also demonstrated focal decreased fast activity in the appropriate sphenoidal-ear channel. Both of these groups did very well post-operatively with respect to their epilepsy. Five patients with large temporal convexity cicatrices antedating seizures all demonstrated lateral temporal primary spike foci without independent secondary foci or focal decreased fast activity and did not do as well post-operatively as the first two groups. Eleven patients had only non-specific changes in the resected temporal lobe and in general did not benefit from surgery. Various combinations of primary and independent secondary spike foci were seen. Only this group demonstrated diffuse or bifrontal spikes during initial EEG recording, and basal mid-line spikes with intravenous thiopentone. Pecilar sharp notched spike were also very common in this group, but not unique to it. Focal decreases in barbiturate-induced fast activity were not noted.


Unmyelinated axons in human ventral roots, a possible explanation for the failure of dorsal rhizotomy to relieve pain. Human ventral roots were examined in the light and electron microscopes. The noteworthy finding is that all roots contain large numbers of unmyelinated axons, and that they make up 27 per cent of the total population of ventral root axons. The function of these unmyelinated axons is not known, but for various reasons it is probable that a significant number are sensory. If so these axons might explain the failure of dorsal rhizotomy to relieve pain, and dorsal root gangionectomy, which would remove both dorsal and ventral root afferents, might be the procedure of choice for those patients where rhizotomy is contemplated.


The tonic ambulatory foot response. A clinical and electromyographic study. Three cases with "pyramidal" symptoms in which standing and/or walking evoked a tonic flexion of the toes and sole of the affected foot, have been studied from the clinical and electrophysiological standpoint. The phenomenon, for which the definition of tonic ambulatory foot respone (TAFR) is proposed, appeared to be triggered by cutaneous stimuli travelling along fibres lying in the range of nociceptive afferents, and to depend on a background of complex sensorimotor patterns of standing posture and/or stepping movements against resistance. The relationship of TAFR to other flexor foot responses described in the literature is discussed.


The pathological anatomy of posthemiplegic athetosis. Disorders of movement after hemiplegia have been described for more than a century, but their pathological anatomy and physiology have remained poorly understood because of ambiguous terminology and incomplete studies. We examined the brains of 5 patients which had been serially sectioned where there had been well documented pure motor hemiplegia acquired in childhood. In 4 patients handicapped by hemiathetosis the main lesion was partial destruction of the caudate nucleus and putamen. In the fifth case, where non-disabling involuntary movements only appeared in later life, there was gliosis of the caudate nucleus and thalamus. Striatal lesions produce involuntary movement disorders if the corticospinal and other major motor tracts are partly intact. We propose that degeneration of the thalamic nuclei receiving striatal efferents (ventralis anterior, ventralis lateralis and centrum medianum), wheter primary or secondary, appears to remove an essential modulating influence on the corticospinal system which can only become manifest if this system is relatively preserved.


Trace element concentration in human brain. Activation analysis of cobalt, iron, rubidium, selenium, zinc, chromium, silver, cesium, antimony and scandium. Up to 60 tissue samples were dissected from 13 human brains in defined regions and were analysed by means of neutron activation analysis for trace element concentration of cobalt, iron, rubidium, selesium, zinc, chromium, silver, cesium, antimony and scandium. It was shown that the variation in concentration of the non-essential elements in the different brain regions was greater than the corresponding variations in concentration of the essential elements. The mean values of Fe and Rb concentrations were higher in the basal ganglia than in the cortex areas. With increasing age the Fe concentration in the human brain was found to increase and that of Rb to decrease. Comparison of the trace element concentration in corresponding areas of the right and left hemispheres showed highly significant positive correlations for the essential elements. The concentration of each of the essential elements Fe, Rb and Zn was also found to differ significantly between defined functional regions. The characteristic distribution of essential trace elements in different areas is discussed in the light of known metabolic functions of these elements.


Symptomatic vertebral haemangiomas.

Symptomatic vertebral haemangiomas. Eight cases of vertebral haemangioma causing spinal cord or nerve root compression are described, together with one other which was not causing organic signs or symptoms. The compression was due to extradural tumour in 5 cases, bony expansion encroaching on the neural canal in 2 cases, and a combination of bony expansion and extradural tumour in one case. When present the classical radiological appearance of accentuated vertical striation or honeycomb pattern is easily recognized, but atypical features, such as apparent pedicular erosion, paravertebral soft tissue mass and bony expansion, may occur, making diagnosis more diffcult. In 2 cases the correct diagnosis was not made before surgery. In one of these there were no plain film changes and in the other a metastasis was considered the more likely diagnosis. In the asymptomatic case the absence of extra-osseous extension was an important factor in excluding any possible clinical significance of the haemangioma. The great importance of pre-operative spinal angiography is stressed.


Transfer of radioactive material between electrically coupled neurons of the leech central nervous system. Intracellular application of tritiated precursors by means of microiontophoresis was performed on nerve cells in isolated segmental ganglia of the leech ventral nerve cord. Incorporation as well as intra- and interneuronal transport were studied by autoradiography after injection of fucose, glucosamine, glycine, leucine, orotic acid and uridine. With several minutes of intraneuronal injection the precursors were incorporated into macromolecules. Depending upon the tracer used, the radioactive material was distributed in a specific pattern over the cell somata and then released into the nerve processes. After application of orotic acid and uridine a transport of radioactive material, presumably RNA, could be observed in the processes of the injected neurons at a distance of about 200-500 mum. Fucose and glucosamine injection resulted in the most extended labeling of the nerve cell processes, indicating a transport rate of about 11 mm/day. When the radiochemicals were injected into one of the two electrically coupled giant nerve cells -- the so-called Retzius cells (Rc) -- a specific labeling not only of the injected Rc but also of the coupled but not injected Rc was found. Injection of protein or glycoprotein precursors into one Rc produced heavy labeling of both Rcs including their processes; a slight labeling of other ganglion compartments was only found after increasing the dosage of the amino acids glycine and leucine. With orotic acid and uridine this interneuronal transfer was confined to the electrically coupled Rc twin. Intracellular injection of one Rc with puromycin followed by injection of amino acids or fucose into the same Rc or into the coupled Rc resulted in an inhibition of precursor incorporation within the puromycin-injected Rc and an exclusive labeling of the coupled Rc, thus indicating that the precursors themselves were transferred. It is suggested that after microiontophoretic application an interneuronal transfer of relatively low molecular weight material takes place, probably across the low-resistance junction through which the Rcs are electrically coupled.


High affinity uptake of GABA in presumed GABA-ERGIC nerve endings in rat brain. After interuption of the striato-nigral pathway uptake of gamma-aminobutyric acid (GABA) in the substatia nigra decresaed with 7 days to a constant level 30-40% of the normal. Concomitantly glutamate decarbosylase (GAD) was reduced to 10%. Hence about two-thirds of the GABA uptake activity in substantia nigra are localised, to the alleged GABA-ergic nerve elements originating from corpus striatum. The lesion resistant part of the uptake is probably not localised in cell bodies or large processes, since it was the same in tissuse prisms as in whole homogenates and crude nerve ending fractions. It was also not influenced by aminoozyacetic acid, which would argue against a localisation in glia. Whereas GAD was recovered mainly in a "heavy" nerve ending fraction, a large proportion of the GABA uptake was situated in a "light" fraction. After hemisections, GABA uptake was reduced to a similar extent in both fractions. It is suggested that whereas GAD is concentrated in nerve terminals, a significant proportion of the GABA uptake may be localised in preterminal axon branches in the substantia nigra. GABA uptake in the dorsal part of the lateral vestibular nucleus was not reduced by interruption of the Purkinje axons from the cerebellar vermis whereas GAD was reduced 50%. This indicates that the reuptake mechanism is not concentrated in the Prukinje axon terminals. In the hippocampus neither GABA nor GAD were reduced by lesions of afferent nerve pathoways, in accordance with previous results showing that in this region GABA producing neurones are intrinsic. The order of ratiols of GABA uptake to particulate GAD activity in different regions was: hippocampus greater than cerebellar cortex greater than substantia nigra greater than dorsal part of lateral vestibular necleus approximately equal to nucleus interpositus. The ratio may relfect the degree of specific localisation of the GABA uptake mechanism to the GABA-ergic structures.


Changes in amino acid influx with Na flow in incubated slices of mouse brain. The uptake of amino acids into slices of adult and newborn mouse brain was studied in relation to Na+ flow. (1) The level of Na+ and K+ incubated slices of brain depends on the ionic composition of the incubation medium. The intracellular levels of Na+ in adult tissue are below, in fetal tissue above, Na+ levels in the medium used. Rapid net flow into or out of the tissue can be achieved by transferring slices into media of higher or lower Na+ content. (2) Under conditions of net Na+ inflow, the influx of all amino acids tested increased; under conditions of net Na+ outflow, the influx of all amino acids decreased, as compared to slices in ionic equilibrium. The absolute levels of Na+ in the tissue under the experimental conditions had little effect on amino acid uptake. The stimulatory effect of Na+ inflow and the inhibitory effect of Na+ outflow could be observed at all developmental stages--in adult, newborn, and fetal tissue. (3) We conclude that ion movements influence metabolite transport; these effects are smaller in the absence of fully developed ion pumps. The direction of the net ion flow does not seem to be the main determinant: active accumulation occurred both in adult and in fetal brain, although the tissue-to-medium Na+ gradients were in opposite directions; in addition, amino acid inflow occurred in the presence of Na+ outflow.


Histamine-n-methyltransferase activity of the nervous system of the chick during development. The developmental course of histamine-N-methyltransferase activity was determined in the chick pineal gland, thalamus, cerebral hemispheres, cerebellum and sciatic nerve from the 13-day embryo to 1-week post-hatching. In each tissue, low levels of enzyme activities were detectable in the 13-day embryo. Thereafter, to the stage of hatching activity rose rapidly in the pineals, thalamus and peripheral nerve. Enzyme activity in the pineals decreased after hatching and remined at a relatively low pre-hatch level in the 7-day chick. In the sciatic nerve and thalamus activity also dropped slightly after hatching. The increase of enzyme activity in the cerebellum and cerebellum hemispheres was very gradual after the 13th day of embryonic stage and maximum activities were obtained only 2 days after hatching. Highest specific activities of the enzyme were detected in the sciatic nerve, pineals and thalamus at each developmental stage. The Km values for histamine and S-adenosyl-l-methionine and the behavior towards certain drugs of the enzyme in the pineals and sciatic nerve did not change significantly during development.


Studies on bursting pacemaker potential activity in molluscan neurons. I. Membrane properties and ionic contributions. Bursting pacemaker potential (BPP) activity of identified molluscan neurons has been studied using cells from Aplysia and Otala. The results presented in this paper indicate that (1) a potassium conductance mediates the hyperpolarizing phase of the BPP; (2) the BPP amplitude is directly dependent on [Na+]0; (3) BPP activity requires the presence of divalent cations and is prevented by C02+ and La3+, but not D-600; (4) the apparent increase in membrane resistance during the depolarizing phase of the Bd can be accounted for by the movement of the membrane potential along the non-linear portion of the I-V curve; and (5) non-linear I-V relations and a minimal effective membrane resistance are pre-requisite to BPP generation. Coupled with recent observations on the presence of an inward current in these cells, the results suggest that the mechanisms underlying the BPP are similar to those proposed to describe the myocardial pacemaker potential: the hyperpolarizing phase is due to activation of a potassium conductance which slowly inactivates, resulting in a gradula deplorization until a voltage-dependent inward current is activated which then leads to an increasingly rapid deplorization and initiation of the burst of spikes. It would appear that Na+ may play the major role in carrying the inward current, although a secondary role for divalent cations cannot be discounted.


Studies on bursting pacemaker potential activity in molluscan neurons. III. Effects of hormones. Vertebrate peptides and hormones have been appled to a number of identified neurosecretory and ono-neurosecretory cells in two molluscan preparations. Active peptide hormones included vasopressin and analogues. Active steriod hormones included aldosterone and hydrocortisone. Peptide effects were present at 10-9 M concentration of peptide, were confined to two neurosecrotory cells and consisted of long lasting changes in the membrane properties of these cells (characterized either by the initiation or potentiation of bursting pacemaker potential activity in these cells). The regulatory changes in membrane properties induced by the peptides were unlike the transient conductance changes produced by conventional neurotransmitters. Steroid effects were observed at 10-6M concentration of steroid and consisted of an increase in membrane potential and conductance which was dependent on the species of divalent cations present. The net effect of peptide activation would be to increase the release of neurosecretory material form the cell terminals, while the net effect of the steroids would be to decrease the release of this material. The results obtained with these invertebrate preparations may serve to describe new forms of cellular communication in the nervous system whereby peptides and steroids modulate electrical activity.


Sustained and transient discharges of retinal ganglion cells during spontaneous eye movements of cat. Discharges of 223 retinal ganglion cells during spontaneous eye movements (saccades) across a stationary grating pattern were studied in chronically prepared cats. Of these 83 showed sustained responses to local differences in luminance of the grating stripes (S-units); 84 showed transient responses to saccades and did not register local differences in luminance (T-units); and 56 showed mixed responses, i.e., transient responses to saccades and sustained firings in response to local luminance (M-units). When tested with diffuse light, 93.9% of the S-units showed either ON-sustained or OFF-sustained responses; 95.2% of the T-units showed either ON-transient, OFF-transient, or ON-OFF-transient responses; and 50% of the M-units showed ON-OFF responses. In the overall responses properties, most S-units corresponded to the X-cells, most T-units to the Y-cells of retinal ganglion cells previously known from acute experiments. Under normal conditions of active eye movements, the major function of the S-units would be to register the differences in luminance in their receptive fields, and subserve the mechansim of form recognition. The major function of the T-units would be to register information related to quick image motion, induced either by eye or object movements, and subserve the mechanism of detecting the dynamic aspects of visual stimuli. The other important functions of the T-units are their possible participation in the afferent routes for two recently proposed mechanisms; one for goal-directed saccades and the other for saccadic suppression. The M-units would possess the functions of both S- and T-units.


Intracranial self-stimulation effects along the route of the nigro-striatal bundle. Recent evidence suggesting a possible dopaminergic nigro-striatal substrate of self-stimulation led  us to map this route for both self-stimulation and stimulus-bound motor effects. The results of 128 electrode placements show that the route of the nigro-striatal projection supports strong self-stimulation effects from the substantia nigra to the ento-peduncular nucleus. Beyond this level, such effects disappear. Our results indicate that the striatum itself is neutral with regard to reinforcement, and suggest that such apparent neutrality cannot be ascribed to motor or other artifacts. These findings require a reappraisal of the hypothesis of a dopaminergic self-stimulation system, although they are not in conflict with the idea that dopaminergic manipulations may affect self-stimulation through some more general regulatory influence on operant responding.


Distribution, metabolism and biological activity of deoxycorticosterone in the central nervous system. Intravenously injected [1,2-3H]deoxycorticosterone (DOC) readily enters all parts of the central nervous system. In intact and eviscerated rats the highest concentration of radioactive label was recovered from areas corresponding to the reticular formation from the brain stem. In addrenalectomized animals, in addition to high brain stem concentration, there was also a marked increase in the uptake of radioactivity in the septum, hippocampus and pituitary. Data from the eviscerated rat point to a uniform distribution of [1,2-3H]DOC in neural tissues and suggest that the higher levels of radioactivity found in the brain stem may be due to a DOC metabolite with the chromatographic characteristics of allo-tetrahydro-DOC, an anaesthetic-type steroid. A decrease in the amplitude of evoked sciatic potentials in brain stem sites but not in the thalamic relay nucleus was observed in 52% of the cases studied, following the i.v. injection of 100-150 mug DOC.


Venous stasis retinopathy. A case report. A case of bilateral carotid occlusion presenting with bilateral venous-stasis retinopathy is presented. The possibility of mistaking this condition for diabetic retinopathy is pointed out and the differentiating features are outlined.


Lacrimal pump pressure patterns.

Lacrimal pump pressure patterns. In most cases the tracings of lacrimal pump pressure patterns may be interpreted, but, should only be used in conjunction with the patient's history and physical examination. A Schirmer and a distraction test should be included. The distraction test is important in order to indicate the magnitude of the lid tension and thereby the integrity of one of the components of the lacrimal pump. A variety of clinical entities were presented and correlation between these entities and their lacrimal pump pressure recordings were made. These pressure patterns evolved from 75 monitored lacrimal drainage problem cases. In rare circumstances the silicone tube could not be inserted where no pathology existed. Stenotic puncta or closed systems cause sharp spikes indicative of high pressure and poor flow. Tightening of the lower lid can significantly increase pressure patterns and increase tear transportation. Tightening of the levator palpebrae superioris may increase pressures in the nasolacrimal system. Monitoring of canalicular and lacrimal pressures is an effective means of determining the patency of a DCR and conjunctival DCR. Nasolacrimal system monitoring is a safe, painless and effective method of evaluating disorders within that system.


An analysis of senile changes in the palpebral fissure. This work was carried out to determine the changes in conformity and in physiological movements of the eyelids with advancing age. Some conceptions fostered in the literature are false and should be discarded. Others have been corroborated. We conclude that the whole lateral canthus becomes lax and drifts medially with age. Therefore, surgical correction should aim at restoring the fixation of the lateral canthus to its youthful position.


Neovascular glaucoma in a university clinic. A comparative survey of 100 consecutive clinic and private glaucoma patients was made in an attempt to determine the nature of these populations. Secondary glaucoma was found four times more frequently in the clinic group. The most common secondary form was neovascular glaucoma. This discouraging entity was reviewed with regard to etiology, pathogenesis, and management.


Stereoacuity and fusional amplitude in foveal suppression. A retrospective study of 100 cases with suppression of one fovea was carried out to discover the relationship between the residual angle of strabismus (if present), the quality of stereopsis and fusional amplitudes. No direct linear relationship was present. However, the following conclusion could be reached. The greatest number of patients with one fovea suppressed who exhibited normal fusional amplitude have a residual esotropia between 5 and 10 prism dioptres. Stereoacuity, as well, tended to be superior in this group. From a practical standpoint, in patients with permanent suppression of one fovea, under binocular conditions, the eradication of any residual tropia, provided it is under 10 prism dioptres, will not improve binocular visual function.


Recurrent T.R.I.C. kerato-conjunctivitis: treatment with tetracycline. Trachoma inclusion conjunctivitis agent infections have a spectrum of clinical presentations which in their more chronic forms are often difficult to recognize. Patients with epithelial erosions of the upper cornea, episodic E.K.C. type subepithelial infiltrates, pannus or micropannus, and with or without lid scarring, merit conjunctival scrapings as an aid to diagnosis. Since the agents which produce these infections have a genital reservoir, as the prevalence of venereal diseases increases, the incidence of this type of keratoconjunctivitis will also increase. If the diagnosis is suspected treatment with systemic tetracycline is effective and relatively safe. Simultaneous treatment of sexual consorts of inclusion conjunctivitis patients and families of trachoma patients should decrease recurrences.


The assessment of near vision.

The assessment of near vision. There are many variables in the physiology of near vision, the end-point criteria, the types available, and the measurements made. I have suggested a hard criterion of the smallest print at the nearest distance. This controls the physiological variables and provides repeatable data for the evaluation of vision, accommodation, and binocular function.


Energy: a useful criterion in analysis of human visual evoked response. Visual evoked responses from a diffuse Xenon flash were studied for seven normal and 14 abnormal subjects. Amplitude and latency measurements of the waveforms were compared with an energy function calculated from the area beneath a curve of the VER voltage squared and summed over the interval from 0 to 125 msec. This energy function proved superior to latency in detecting asymmetrical responses and similar to latency in detecting symmetrically reduced waveforms.


Melanin deposition in the hyphae of a species of Phomopsis. Hyaline hyphae of Phomopsis become pigmented when exposed to short periods of light. Pigment was deposited in the form of melanin granules both within the cell wall and within mucilaginous excrescences that were developed irregularly over the hyphal surface. Analysis of the pigment showed it to have properties similar to that of "Dopa" melanin and to pigments previously isolated from fungal cell walls. Lysis of both hyaline and pigmented hyphal walls by means of lytic enzymes was minimal. It is suggested that the major role of melanin in this fungus is the protection of cellular organelles from harmful ionizing radiations.


An experimentally pathogenic Bacillus species. II. The pathogenicity of the organism for mice. The pathogenic effects produced in mice by intraperitoneal (i.p.) injection of a Bacillus species (OSU 372) are presented. This organism belongs to group 3 of the genus, and members of this group have not heretofore been shown to be pathogenic for mice even under experimental conditions. However, this organism is capable of producing a fatal involvement in doses which are not considered to be overwhelming. The mean lethal dose (LD50) of the organism for 20-25 g mice by the i.p. route is about 1 times 10-8 bacteria/mouse. A rapid drop in body temperature along with severe dehydration were noted in infected animals, and hematologic studies indicated that leukopenia and hemoconcentration also occurred. Although a transient septicemia developed, the bacteria could not be recovered from the tissues of fatally infected mice after a certain point in time. Results prevented indicate that the animals died of hypovolemic shock. A possible parallel with human bacillary infection is drawn.


Changes in cutaneous flora after wet occlusion. Aerobic flora from wet-occluded forearms of six volunteers was sampled the day before treatment, on the 3rd day when dressings were removed, and daily, when possible, for 8 days thereafter. Erythema was not present. All bacterial colonies appearing on appropriate dilution plates were identified with the aid of a replica-plating technique. Flora of each individual increased to over 10-4 colony-forming units/cm2 as a result of wet-occlusion, but counts rapidly fell by about 10-2 units once dressings were removed. Although similar types of bacteria were found on all subjects, the composition of each individual's flora during the recovery response appeared to be unique. Enterobacteriaceae were found on half the subjects with Enterobacter aerogenes being the most successful colonizer. Besides the expected presence of Baird-Parker Staphylococcus subgroup II, high  numbers of subgroup IV and some colonies of subgroup III were also observed. Almost all cutaneous diphtheroids were lipophilic and lipolytic.


Characterization of branched and unsaturated fatty acids in Mycobacterium vaccae strain JOB5. After growth of Mycobacterium vaccae strain JOB5 on acetate or propane, the cellular fatty acids were isolated and identified by a combination of gas-chromatographic, mass-spectral, and chemical means. The fatty acids ranged from C12 to C19 and were a mixture of saturated, monounsaturated, and methyl-branched components. The double bond was in the delta9 position in the C15 to C18 unsaturated acids. The single methyl branch was located on the C10 position of Br-C17, Br-C18, and Br-C19 fatty acids. Branched-chain synthesis occurs at the expense of an unsaturated precursor fatty acid; the double bond serves as the site of methylation. Results suggest that S-adenosylmethionine is the methyl donor involved.


Fungal fimbriae. II. Their role in conjugation in Ustilago violacea. During conjugation in the anther smut fungus Ustilago violacea cells of opposite mating type first pair tightly and then develop a conjugation tube or bridge between them. The cells of both mating types are covered in long fine hairs or fimbriae, some of which appear to end in knobs. Experiments involving enzyme treatments of the cell surface indicate that these fimbriae do not play an essential role in cell pairing, instead pairing seems to be initiated when one or both mating types produce amorphous masses of alpha-amylase-sensitive material. Electron micrographs, enzyme and inhibitor studies, and experiments using restrictive temperatures suggest, however, that fimbriae may be essential for the later stages of conjugation i.e. development of the conjugation tube. If so, it is suggested that they may permit the exchange of macromolecules between the conjugating cells, initiating localized wall-softening and wall-breakdown.


A large new Streptococcus bacteriophage. Phage VD13 possesses a large, elongated head and a long, non-contractile tail. It is active on group D streptococci and contains double-stranded DNA. The phage produces several rare kinds of head malformations, notably polymorphic mottled structures and giant heads which probably contain DNA.


Hemoglobinopathies in the Hamilton region. II. Thalassemia traits and iron therapy. Between July 1973 and July 1974 all adult patients with hypochromic anemia and a mean corpuscular volume of 75 mum3 or less were screened for hemoglobinopathies. Of the 490 patients 105 had beta-thalassemia trait, 11 had alpha1-thalassemia trait, 4 had hemoglobin Lepore trait and 1 had hemoglobin H disease. Of 48 inpatients whose charts were reviewed 19 had been on oral iron therapy and 7 of them had been given iron intramuscularly. Of 27 outpatients interviewed 10 had been on intermittent iron therapy for 18 months or more; 4 had been given at least 1 g of intramuscular iron. Iron deficiency was not documented in any of these patients. Iron deficiency should be diagnosed by means other than the presence of a hypochromic picture in the peripheral blood before iron therapy is instituted, particularly in communities with a large population of Mediterranean or South-East Asian origin.


Spontaneous pneumothorax: outpatient management with intercostal tube drainage. In a series of 104 episodes of pneumothorax 75 percent of episodes were managed successfully on an outpatient basis by observation (23.1 percent) or by intercostal tube drainage using a flutter valve (51.9 percent). The patients for whom this treatment was not successful were admitted to hospital; 17 of them (16.3 percent of 104) were treated surgically. Bleb suturing with a stapling device and dry sponge abrasion of the pleura was the operation of choice.