A case report of fowl plague keratoconjunctivitis. A case of human fowl plague keratoconjunctivitis occurred after accidental laboratory exposure. The conjunctivitis was characterised by follicle formation and a mucopurulent discharge, and ran a self-limiting course over two weeks. The keratitis was of an unusual type and consisted of small intraepithelial opacities, which appeared after one week and resolved completely over the next three weeks. The infection, confirmed by viral culture, was produced by Dutch strain (Hav 1 Neq 1) of fowl plague virus.


Ticarcillin in the treatment of experimental pseudomonas keratitis. In rabbits with experimental Pseudomonas aeruginosa keratitis the subconjunctival administration of either ticarcillin or gentamicin significantly ameliorated the keratitis and reduced the number of organisms to be found in the cornea. The use of these drugs in combination had an additive effect in vitro studies and a significantly more beneficial effect on the clinical disease than the use of either drug alone.


Conjugated polyene fatty acids on fluorescent probes: spectroscopic characterization. This paper is the first in a series which extends introductory studies of parinaric acid and its phospholipid derivatives as membrane probes (Sklar, L.A., Hudson, B., and Simoni, R.D. (1975), Proc. Natl. Acad. Sci. after U.S.A. 72, 1649; (1976), J. Supramol. Struct. 4, 449). Parinaric acid has a conjugated tetraene chromophore and exhibits many spectroscopic properties common to linear polyenes. Its absorption spectrum is characterized by a strong near-ultraviolet transition with vibronic structure, which is strongly affected by solvent polarizability. The fluorescence emission occurs at considerably lower energy than the absorption and the wavelength of the emission is nearly independent of the solvent. The fluorescence quantum yield and lifetime are strongly affected by temperature and solvent. These spectral features are interpreted in terms of an excited electronic-state order such that a weak transition occurs at longer wavelengths than the strongly allowed transition which dominates the absorption. The sensitivity of the fluorescence quantum yield an lifetime to environment is shown to be due primarily to variations in the nonradiative rate, although changes in the radiative rate constant are also observed and interpreted. The absorption spectrum (epsilon max greater than 65 000) is in the 300-320-nm range, a region relatively free of absorption due to intrinsic biological chromophores. Shifts of several nanometers are characteristic of different environments. These shifts are compared to similar effects observed for a series of diphenylpolyenes for which new data are given and are correlated using a simple but adequate theory of solvent shifts. The intrinsic (or radiative) fluorescence lifetime is near 100 ns in a wide variety of environments. This is much longer than the intrinsic lifetime calculated from the absorption spectrum and strongly supports the proposed excited-state order.


Conjugated polyene fatty acids as fluorescent probes: synthetic phospholipid membrane studies. The preparation of polyene fatty acid membrane probes cis- and trans-parinaric acid and parinaroylphosphatidylcholines and their use in studies of several one- and two- component lipid systems are described. The fluorescence quantum yield of trans-parinaric acid in dipalmitoylphosphatidylcholine at 20 degrees C is approximately 0.3; the quantum yield in aqueous solution is negligibly small. Thermal-phase transitions in single-component phospholipid dispersions are monitored with absorption and fluorescence excitation peak position, fluorescence intensity, lifetime, and polarization. The transition temperatures observed are consistent with previous determinations. Shifts in the absorption peak position are related to the bilayer expansion as it undergoes the gel to liquid-crystalline transition, while fluorescence depolarization provides semiquantitative information concerning molecular motion of the probe in the bilayer. A long fluorescence lifetime component is observed for parinaric acid in the solid phase (up to 50 ns), and a short lifetime component is observed (ca. 5 ns) in the fluid phase of dipalmitoylphosphatidylcholine; both lifetime components are observed in the transition region. In most phospholipids, cis-parinaric acid detects the melting transition at about 1 degree C lower than trans-parinaric acid. Partitioning experiments involving mixed populations of phospholipid vesicles show that trans-parinaric acid preferentially associates with solid-phase lipids, while cis-parinaric acid shows a more equal distribution between solid and fluid lipids. The binding of cis-parinaric acid to dipalmitoylphosphatidylcholine at 25 degrees C is described as partitioning of parinaric acid between lipid vesicles and the aqueous phase with a partition coefficient of 5 X 10(5). Several rates are observed in the binding process which are interpreted as rapid outer monolayer uptake and a much slower process of interlamellar exchange. The phase diagram of the binary lipid mixture dipalmitoylphosphatidylcholine-dipalmitoylphosphatidylethanolamine has also been examined and found to be essentially identical to the one constructed using a nitroxide probe.


The progeny of rabbit articular chondrocytes synthesize collagen types I and III and type I trimer, but not type II. Verifications by cyanogen bromide peptide analysis. The radioactive collagens synthesized by the fourth subculture progeny of rabbit articular chondrocytes were extracted and purified after limited pepsin digestion by neutral and acid salt precipitation. In order to identify the different types of collagen present, denatured collagen chains were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 5% gels, electrophoretically eluted, and cleaved with cyanogen bromide, and the resultant peptides were fractionated by a new sodium dodecyl sulfate electrophoresis system (tris(hydroxymethyl)aminomethane-borate buffer, 15% gels). Comparison of these separate peptide profiles with those from alpha1(I) and alpha1(III) collagen chains permitted the unambiguous identification of these chains in the radioactive collagen synthesized by chondrocytes. Although cartilage slices predominantly synthesized alpha1(II) chains, only alpha1(I) chains were made by cells in fourth subculture. A large fraction of these alpha1(I) chains could not be accounted for by the presence of type I collagen. While in a native, triple-helical conformation, some of these extra alpha1(I) chains were completely separated from type I collagen by their solubility at pH 8.0 in 2.6 M NaCl and therefore identified as [alpha1(I)]3, type I trimer. In addition to type I collagen and type I trimer, these chondrocyte progeny also synthesized type III collagen and two new collagen chains, X and Y. Each collagen type was further characterized by carboxymethylcellulose chromatography and its distribution between the medium and the cell layer. These findings support the idea that cultured chondrocytes assume a collagen phenotype similar to that of their undifferentiated mesenchymal cell precursors.


The complete amino acid sequence of the major component myoglobin of dwarf sperm whale (Kogia simus). The complete amino acid sequence of the major component myoglobin from the dwarf sperm whale, Kogia simus, was determined by specific cleavage of the protein to obtain large peptides which are readily degraded by the automatic sequenator. Three easily separable peptides were obtained by cleaving the protein at its two methionine residues, and five peptides were obtained from the methyl acetimidated protein by cleavage with trypsin at the four arginine residues. Sequenator analysis of these fragments and the apomyoglobin provided over 80% of the covalent structure of the protein. The remainder of the primary structure was determined by further digestion of the two larger cyanogen bromide fragments with trypsin and staphylococcal protease. To reconfirm many of the substitutions found in this protein, the apomyoglobin was treated with 1,2-cyclohexanedione, and the resulting arginine protected protein was cleaved at its lysine residues with trypsin. This myoglobin differs from that of the sperm whale at 6 positions, and from the other cetacean myoglobins at about 16 positions. The appearance of a histidine residue at position 35 has no precedent in any myoglobin. The substitutions seen at positions 21, 51, and 132 are unique to date for cetacean myoglobins.


A 13C nuclear magnetic resonance study of gel-forming (1 goes to 3)-beta-d-glucans. Evidence of the presence of single-helical conformation in a resilient gel of a curdlan-type polysaccharide 13140 from Alcaligenes faecalis var. myxogenes IFO 13140. A 13C nuclear magnetic resonance (NMR) study of a resilient gel of a curdlan-type polysaccharide 13140, a (1 goes to 3)-beta-D-glucan, from Alcaligenes faecalis var. myxogenes IFO 13140, was performed in an effort to understand the gel structure. It was found that very broad 13C resonance peaks of line widths ca. 150 (C-1-C-5) and 50 HZ (C-6) are able to be seen in the gel state. Because of a fivefold increase of the peak intensities with respect to those in an aqueous suspension by gelation, these peaks are unequivocally ascribed to certain regions relevant to the gel structure. Those 13 C NMR peaks, however, account for only 20-30 (as viewed from C-1-C-5) and 60% (from C-6) of the total gel, the peak areas of the rest being lost. With respect to those of the disordered low molecular weight acid degraded fraction, fraction II, and laminaran, downfield displacements of C-1, C-3, and C-4 signals are found to take place by amounts of 2.8, 3.2, and 0.9 ppm, respectively, while the remaining peaks (C-2, C-5, and C-6) are unchanged. In view of similar differences of chemical shifts between cyclodextrins and linear (1 goes to 4)-alpha-D-glucans (Colson, P., Jennings, H.J., and Smith, I.C.P. (1974), J. Am. Chem. Soc. 96, 8081-8087), the observed downfield 13C shifts of the glucosidic bonds in the gel are explained by the presence of the fixed conformation of the preferred dihedral angles, in which internal rotations around the glucosidic bonds are not allowed. Combined with the results of theoretical prediction and the downfield shift of C-4 signals, which is consistent with the presence of an O-4'...O-5 intramolecular hydrogen bond, the observed 13C peaks are ascribed to a region of single helical conformation, whereas the peak-loss portion of 13C NMR signals (70-80 and 40% from C-1 and C-6, respectively) is presumably ascribed to the multiple-helical junction zones for the gel structure and their vicinities. The variation of the line widths as well as the peak positions is also found to take place by stepwise addition of NaOH (greater than 0.22 M). The onset of the conformational transition, helix to random coil, is in good agreement with the change of viscosity, specific rotation, optical rotatory dispersion, and absorption maximum shift by complex formation with Congo Red reported by Ogawa et al. (Ogawa, K., Watanabe, T., Tsurugi, J., and Ono, S. (1972), Carbohydr. Res. 23, 399-405).


Nucleoside adducts from the in vitro reaction of benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide or benzo[a]pyrene 4,5-oxide with nucleic acids. The covalent binding of benzo[a]pyrene 4,5-oxide and benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide isomer I and isomer II to nucleic acids in aqueous acetone solution has been investigated. Benzo[a]pyrene 4,5-oxide reacted preferentially with guanosine residues. On the other hand, benzo[a]pyrene-7,8-dihydrodiol 9,10-oxide isomer I and II reacted extensively with guanosine, adenosine, and cytidine residues. Time course studies showed that the reactivity of isomer I or isomer II with homopolyribonucleotides followed the order poly(G) greater than poly(A) greater than poly(C). Alkaline or enzymatic hydrolysis of the modified nucleic acids and subsequent chromatography on Sephadex LH-20 columns yielded benzo[a]pyrene-nucleotide adducts. These were enzymatically converted to the corresponding nucleosides which were resolved into several distinct components by high-pressure liquid chromatography. Evidence was obtained for the presence of multiple nucleoside adducts of guanosine, adenosine, cytidine, deoxyguanosine, deoxyadenosine, and deoxycytidine. The HPLC profiles of adducts formed with isomer I were different from the corresponding profiles of adducts formed with isomer II. Structural aspects of these nucleoside adducts are discussed.


Equilibration of fucosyl glycoprotein pools in HeLa cells. The pool sizes, label equilibration times, and specific radioactivity relationships of fucosyl glycoproteins and precursors have been examined in exponentially growing HeLa S3 cells (generation time about 23 h) using a quantitative radioisotopic approach. The specific radioactivity of the precursor GDP-fucose (pool size 0.52 +/- 4% nmol/10(7) cells) equilibrates with radioactive fucose in the medium in about 1 h. 10(7) cells contain 5.3 +/- 16% nmol of glycoprotein fucose of which 96-98% resides in or on the cell surfaces and is equilibrated isotopically within 22 h of labeling; 2%  or less is in an internal pool, some of which is precursor to plasma membranes and some of which is released as soluble glycoprotein directly to the medium without random mixing with the plasma membrane glycoprotein. Because we cannot rule out the presence of internal free fucose, 2% of the total glycoprotein fucose could be in the degradative pathway being recycled internally before release of free fucose. In rate terms and in a particular culture where 10(7) cells contained 4.4 nmol of glycoprotein fucose, a total of 11.1 nmol of glycoprotein fucose is synthesized per generation (9-11% of the total cell glycoprotein fucose/h). Of this, 2.5 nmol of glycoprotein fucose per generation is released directly into the growth medium without mixing with the plasma membrane glycoprotein fucose. The small internal pool feeding glycoprotein fucose to the plasma membrane does so at the rate of 8.6 nmol/10(7) cells per generation, 4.2 nmol per generation of which, after mixing with the plasma membrane glycoprotein fucose, is ultimately released into the growth medium, 75-80% as free frucose. This release process is independent of cell density and the presence of serum in the growth medium.


Differential effects of puromycin on the incorporation of precursors of rhodopsin in bovine retina. Bovine retinas incubated in vitro sustain the synthesis of opsin and rhodopsin as monitored by the incorporation of labeled leucine, mannose, and glucosamine. Puromycin, an inhibitor of protein synthesis, effectively blocks the incorporation of leucine and mannose into opsin and rhodopsin of rod outer segments. However, the incorporation of glucosamine into opsin and rhodopsin is not immediately blocked. Instead, it continues for a time suggesting not only core oligosaccharide synthesis but also the secondary glycosylation of a pool of preformed opsin which is thought to be transiently accumulated in the photoreceptor Golgi complex. Galactose, not normally found in rhodopsin, is also incorporated into both opsin and rhodopsin. This incorporation appears to be completely insensitive to puromycin, suggesting that it may occur in the rod outer segments involving only preexisting glycoproteins.


Degree of exposure of membrane proteins determined by fluorescence quenching. Quaternary salts of 4-picoline are shown to act as efficient quenchers of tryptophan fluorescence in membrane proteins. Fluorescence quenching determinations of sarcoplasmic reticulum membranes from rabbit muscle and of human erythrocyte membranes of different cholesterol to phospholipid mole ratios (C/PL) were carried out with quaternary picolinium salts in phosphate-buffered saline (PBS) and in 2,2,2-trifluorethanol (TFE)-water 2:1 (v/v), where the membrane is presumably completely disintegrated. In both solvent systems, the tryptophan quenching characteristics were typical of heterogenous systems and were analyzed as such. The ratio of the fraction of fluorescence intensity available for quenching with N-methylpicolinium perchlorate in PBS and in 2:1 TFE-water, (formula: see text), was taken as an index for the bulk degree of exposure of the membrane proteins to the aqueous surrounding. This value was found to increase with C/PL which is in line with the notion that increase in lipid microviscosity results in increase of exposure of membrane proteins. Analogous experiments were performed with N-hexyl- and N-benzylpicolinium, which can quench tryptophyl residues in both the aqueous phase and the hydrocarbon-water interface, and with N-hexadecylpicolinium which is dissolved in the membrane lipid layer and acts mostly as a static quencher of tryptophan at the hydrocarbon-water interface. With these quenchers the complementary indices (formula: see text) and (formula: see text), which represent the fraction of the protein mass located in the hydrocarbon-water interface and in the hydrocarbon layer, respectively, could be semiquantitatively resolved.


Quenching of the tyrosyl and tryptophyl fluorescence of subtilisins Carlsberg and Novo by iodide. The tyrosyl and tryptophyl fluorescence of diisopropylphosphorylsubtilisins Carlsberg and Novo, respectively, is quenched efficiently by I- but is not significantly affected by Cs+. The I-quenching data were analyzed using a modified Stern-Volmer treatment (Lehrer, S.S. (1971), Biochemistry 10, 3254), yielding values for the effective fraction of accessible protein fluorescence of 90-95 and 88-92% for the tyrosyl and tryptophyl emission of diisopropyl-phosphorylsubtilising Carlsberg and Novo, respectively. Similar values were obtained pH 5 and 7. The effective collisional quenching constant depends on pH in a manner suggesting the participation of protein surface charge in the quenching mechanism. Significant singlet energy transfer (efficiency = 0.52) from tyrosyl to tryptophyl residues was inferred from the excitation spectra of diisopropylphosphorylsubtilisn Novo. The very low efficiency of energy transfer to Trp-113 in diisopropylphorphorylsubtilisin Carlsberg suggests that Trp-105 and Trp-241 are the acceptors of tyrosyl emission in the homologous Novo enzyme. The unusually low quantum yield of Trp-113 in diisopropylphosphorylsubtilisin Carlsberg together with the tryptophyl fluorescence quenching behavior of the Novo enzyme suggests that this residue is "buried" and in accessible to quenching in both enzymes. The tyrosyl quenching behavior of diisopropylphorphorylsubtilisin Carlsberg is consistent with the high degree of solvent exposure of aromatic residues evident in the X-ray model of subtilisin Novo.


Antibody synthesis stimulation by vitamin A in chickens. The effect of vitamin A on the immune response of chickens on primary and secondary immunization has been studied. It is demonstrated that the antibody content in chickens depends on the dose of vitamin A in the diet. In chickens fed a high dose of vitamin A in the diet the antibody content in the serum is 2 to 5 times as high as in chickens which have not been given vitamin A. An additional dose of vitamin A administered perorally any day following reimmunization increases the content of antibodies; the effect is most pronounced when vitamin A is given on the third day after reimmunization. In the period of maximal synthesis of antibodies (during the fourth day after reimmunization) labelled retinol is incorporated into the spleen of reimmunized chickens 13 times as actively as in the spleen of control chickens. In the in vitro experiments with antibody synthesis by spleen cells it has been demonstrated that addition of retinyl palmitate to the incubation medium enhances antibody synthesis during incubation. Addition of retinyl palmitate to the spleen cells of vitamin A-deficient chickens restores the synthesis up to the level observed in the control cells. The suggestion is discussed that the effect of vitamin A on the immune process is realized at the level of immunoglobulin synthesis.


Stimulation of sea urchin DNA polymerase by protein factors. II. Formation of active complex in DNA polymerase reaction. A protein factor, SF I, which stimulated DNA polymerase activity severalfold was purified from nuclei of sea urchin embryos by phase separation, ammonium sulfate fractionation, DNA-cellulose, CM-cellulose and hydroxyapatitecolumn chromatography and gel filtration. The molecular weight of SF I was about 220 000, the S20,W value was about 8.5 and the isoelectric point was determined to be pH 5.1. In the presence of SF I,V of the DNA-polymerizing reaction was increased and Km values for the substrates of this reaction were not changed. Addition of polyamines increased the rate of stimulation. ATP which was required for stimulation could be substituted by other ribonucleoside triphosphates. SF I, nuclear DNA polymerase and ATP seemed to form an active complex, and in the complex, ATP was found to have been converted to AMP and inorganic pyrophosphate.


The photodynamic modification of DNA by hematoporphyrin. Illumination (white light: 300-750 nm) of DNA in the presence of hematoporphyrin (less than or equal to 5-10 (-4) M) results in selective degradation of the guanine moiety. DNA so illuminated exhibits physical chemical properties (lowered sedimentation coefficients, lower temperatures of helix-coil transitions, increased buoyant density values) consistent with single-chain scissions (and the generation of single-stranded regions) which presumably are secondary to the photodegradation of the guanine residue. Illumination of DNA in the presence of low levels of hematoporphyrin (greater than or equal to 2.5 - 10 (-4) M) results in a biopolymer exhibiting all of the physical properties described above with the exception of a lowered sedimentation coefficient; on the contrary such DNA is aggregated. Of the four usual deoxynucleosides irradiated in the presence of hematoporphyrin, only deoxyguanosine is destroyed.


Structure of animal mitochondrial DNA (base composition, pyrimidine clusters, character of methylation). Base composition, content of pyrimidine isopliths and methylation degree of mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) from various vertebrates and protozoon Crithidia oncopelti have been studied. mtDNAs from mammals (ox, rat) do not differ in fact in the G + C content from the respective nDNA. The G + C content in mtDNA from fishes (sheat-fish) and birds (duck, chicken) is 1.5--2.5 mol % higher than in the respective nDNA. Kinetoplast DNA (kDNA) from Crithidia oncopelti (G + C = 42.9 mol %) differs significantly in base composition from nDNA (G + C 51.3 mol%). All the mtDNA and kDNA studied differ from the respective nDNA by a lower degree of pyrimidine clustering. The amount of mono- and dipyrimidine fragments in mtDNA is more than 30 mol %, whereas in nDNA it does not exceed 23 mol %. The quantity of long pyrimidine clusters (hexa- and others) is 2--4 times lower in mtDNA than in nDNA. The lower degree of clustering of pyrimidine nucleotides seems to be a specific feature of all the mtDNA studied. This may be indicative of common traits in the organization and origin of mtDNA. All mtDNA of vertebrates contain 5-methylcytosine as "minor" base (1.5--3.15 mol %) and surpass by 1.5--2 times the respective nDNA in the methylation degree. It has been found that in animals mtDNA is species specific as far as the 5-methylcytosine content is concerned. mtDNA of beef heart differs significantly from nDNA in the mode of 5-methylcytosine distribution in pyrimidine isopliths, which may indicate that methylation specificity of nuclear and mitochondrial DNA is not the same. In mitochondria and nuclei of rat liver certain DNA-methylase activity has been detected, which provides in vitro the methylation of cytosine residues both in homologbous DNA and various heterologous DNAs. Specificity of methylation in vitro of cytosine residues in one and the same heterologous DNA from Escherichia coli B with nuclear and mitochondrial enzymes is different. Mitochondrial enzyme methylates cytosine residues chiefly in mono-, whereas nuclear enzyme, in di- and tripyrimidine fragments.


Conformational states of 18 S rRNA from HeLa cells detected by gel electrophoresis. Gel electrophoretic analysis has been used to study some of the conformational properties of 18 S rRNA from HeLa cells. After isolation and treatment by heat or denaturants, the mobility of the RNA in 3% acrylamide/0.5% agarose composite gels was found to be altered; often there were two or more distinct components. Several of these forms could be interconverted by heating to 40 degrees C or by use of dimethyl sulfoxide. Although these comformers were easily distinguishable (by their electrophoretic properties), their melting profiles were very similar. Only a single electrophoretic form was detected when electrophoresis was performed at 40 degrees C. The conformational differences observed are probably derived by variations in base pairing and stacking that result when the parent form is cooled.


The heterogeneous nuclear RNA of chicken erythroblasts. Heterogeneous nuclear RNA (hnRNA) from chicken erythroblasts has a modal molecular weight of 1.6 -10(6) in 99% dimethylsulfoxide. When erythroblasts are labeled continuously with [14C]uridine, nuclear RNA is labeled as a single kinetic component with a half-life of 18 min. After a 10--20 min lag, label appears in cytoplasmic RNA at about 1% of the initial rate of total RNA synthesis. Of the hnRNA sedimenting faster than 28 S ribosomal RNA in both an aqueous sucrose gradient and a subsequent fructose gradient in 99% dimethylsulfoxide, about one-third is polyadenylated, although only about one in 2000 (i.e. about four molecules per cell) contain a globin messenger sequence. The hnRNA of erythroblasts isolated from 5.7- and 11-day chick embryos have the same content of globin messenger sequences as erythroblaasts from anemic adults.


Isolation and comparison of ribothymidine-lacking tRNAs of fetal, newborn and adult bovine tissues. Although ribothymidine (rT) is the most common methylated nucleoside in tRNA, a wide variety of bovine tissues have now been found to contain a class of tRNAs which totally lack rT and have an unmodified uridine in its place. The tissues studied include bovine brain, kidney, liver, thymus and testicles from adult, newborn and fetal stages. The class of tRNA was detected by its ability to be methylated with Escherichia coli rT-forming uracil methylase with radioactive S-adenosyl-L-methionine as the methyl donor. In each case rT was shown to account for at least 95% of the methylated products produced. In vitro methylated tRNA populations were compared by fractionation of double-labeled tRNAs on RPC-5 columns. Three major methyl-accepting tRNA peaks were found for all mammalian tissues studied. The level of methyl acceptance in these peaks was found to vary considerably between tRNAs of different tissues. A major difference in the methyl-accepting tRNA populations of bovine liver and calf thymus was observed. Little similarity was found in the rT-lacking class of tRNAs of bovine liver and wheat germ. Three members of the rT-lacking class of bovine liver tRNA were isolated and found to be two species of valine tRNA and one species of threonine tRNA. All three tRNA's completely lacked rT and could be quantitatively methylated with E. coli uracil methylase.


The role of arginine residues at enzyme active sites. The interaction between guanidinium ions and p-nitro-phenyl phosphate and its effect on the rate of hydrolysis of the ester. In alcoholic solutions a relatively strong complex forms among two guanidinium ions and one p-nitrophenylphosphate dianion. The effect of this complex formation on the hydrolysis of the ester is to lower the rate by a factor of 4 in solutions containing 1 M guanidine hydrochloride when compared with solutions of the same total ionic strength containing no guanidinium ion. It is therefore suggested that, for the enzymatically catalyzed hydrolysis of phosphate compounds going via the formation of a metaphosphate intermediate, the role of any arginine residues at the active site is primarily one of binding and positioning the substrate.


Pyruvate kinase isozymes in cells isolated from fetal and regenerating rat liver. There are at least three major mammalian isozymes of pyruvate kinase (ATP : pyruvate 2-O-phosphotransferase, EC 2.7.1.40), designated K4, L4, and M4. Whereas parenchymal cells from adult rat liver contain only the type L isozyme, parenchymal cells isolated from fetal and regenerating liver were found to synthesize both the K4 and L4 isozymes. A small amount of K-M hybrid was seen in regenerating liver, but there were no detectable M-L or K-L hybrids. Thus, it appears that type L pyruvate kinase is not synthesized at the same time in the same liver cell with either of the other two isozymes. The intermediate electrophoretic bands seen with homogenates of whole fetal liver, and in some earlier work attributed to either hybrid isozymes or to the presence of M4, are contributed by nonparenchymal cells which, in the fetus, are largely hemopoietic. These additional bands of pyruvate kinase are electrophoretically and immunologically similar to the pyruvate kinase isozymes found in adult erythrocytes. The results reported here suggest a very rigorous control in the synthesis of K4 and L4 isozymes in parenchymal cells of both fetal and regenerating liver as opposed to developing neurons and glia, where the shift from synthesis of type K to type M subunits appears to occur gradually and results in the production of substantial amounts of hybrid isozymes.


Fluoride inhibition of inorganic pyrophosphatase. II. Isolation and characterization of a covalent intermediate between enzyme and entire substrate molecule. A presumed pyrophosphoryl-enzyme intermediate of the reaction catalyzed by bakers' yeast inorganic pyrophosphatase pyrophosphate phosphohydrolase, EC 3.6.1.1) has been isolated using fluoride-mediated inactivation of the enzyme during catalysis. The analysis of the F--inactivated pyrophosphatase revealed the presence of one molecule of PPi and one atom of fluoride per active site. The incubation of the inactivated enzyme at 25 degrees C and pH 7.2 resulted in gradual recovery of catalytic activity and concomitant removal of PPi by a first-order reaction with tau1/2 of 1 h. The digestion of the F--treated pyrophosphatase with pepsin yielded phosphorous-containing peptides, which were reduced with NaBH4 and gave homoserine and homoserine lactone after acid hydrolysis. This suggests that the PPi residue is linked to the protein through a bond of an acyl phosphate type involving the beta-COOH function of aspartic acid. Together with the results of the kinetic studies of fluoride inhibition of pyrophosphatase reported in accompanying papers, these findings strongly indicate that the enzyme-substrate compound stabilized by fluoride is a transient of the catalytic reaction.


Fluoride inhibition of inorganic pyrophosphatase. III. Dependence on the nature of substrate and metal ion cofactor. Studies of fluoride interaction with  bakers' yeast inorganic pyrophosphatase (pyrophosphate phosphohydrolase, EC 3.6.1.1) in the presence or absence of enzyme-catalyzed reactions have revealed pronounced specificity of this inhibitor. It was found that the inhibition of enzymic hydrolysis of PPi, ADP, ATP and tripolyphosphate in the presence of Zn2+ and Mn2+ at pH 6.5 is not time dependent and by far less extensive as that observed for the Mg2+-stimulated cleavage of PPi (apparent Ki values differ by three orders of magnitude). Addition of Ca2+ to the latter reaction decrease proportionally the activity of the enzyme and the rate constant for the binding of fluoride to it, which indicates that the enzyme-substrate complexes containing both Mg2+ and Ca2+ are inert in the reaction with fluoride. Preincubation of pyrophosphatase with NaF and various metal cations and substrates, in conditions where the enzyme does not act as a catalyst, does not affect its activity compared to controls lacking fluoride. The results are consistent with the proposed mechanism  of mutual hindrance of substrate and fluoride release from the active site of pyrophosphatase.


Molecular orbital studies on the conformation of the phosphopantetheine moiety of coenzyme A. Conformational study on phosphopantetheine shows that this compound has an intrinsic tendency to adopt a multitude of conformations which contain hydrogen bonds involving the sulphydryl, hydroxyl, carbonyl and amide groups. The sulphydryl group may form a hydrogen bond with the C(7') = 0 carbonyl group, the latter being also involved in hydrogen bonding with the N(4')-H GROUP. All these hydrogen bondings occur for different conformations around the backbone. The N(7')-H and C(4') = 0 groups are not involved in hydrogen bonding. It is also found that a strong interaction occurs between N(4')-H and 0-3' which is responsible for a rigid conformation around the C(3')-C(4') and C(3')-0(3') bonds. As far as the phosphate group is concerned the results show that this group may interact with the 0(3')-H hydroxyl group to form hydrogen-bonded rings of different sizes. A six-membered ring formed by hydrogen bonding between 0(3')-H and 0-1' appears more favorable than an eight-membered ring involving an anionic oxygen instead of an ester oxygen related to the phosphate group.


Activation and inhibition of the phosphoglycerate kinase reaction by ATP. A kinetic study for understanding how the phosphoglycerate kinase (ATP: 3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) reaction discriminates between MgATP2- and ATP-4 is presented. The results show that in contrast to MgATP2-, ATP4- is competitive with 3-phospho-D-glycerate. ATP4- binds to the free enzyme as an inhibitor. When binding to the enzyme-MgATP2- (3-phospho-D-glycerate) complex, ATP4- acts as an activator.


Reactivity of neonatal canine aortic strips. To determine whether vascular reactivity in the young animal is significantly different from that of the mature animal, helical strips of thoracic aorta were suspended in an organ bath of Krebs bicarbonate solution, and isometric tension was measured. The effects of norepinephrine, phenylphrine, tyramine, and KCl were studied. The neonatal vascular strips were less sensitive than adult strips to the adrenergic agents, but responsiveness to KCl was comparable in both groups.


Gel filtration of hyperbilirubinemic sera through Sephadex G-25 and Sephadex LH-20 for the detection of 'free' non-albumin-bound unconjugated bilirubin. Hyperbilirubinemic model sera with different bilirubin/albumin (Br/Alb) molar ratio, containing unconjugated bilirubin only, and containing various fractions of conjugated and unconjugated bilirubin, were simultaneously run through small columns packed with Sephadex G-25 and Sephadex LH-20. The bilirubin  adsorbed by the gel was detected by a diazo-reagent and also recovered with chloroform. The amount of bilirubin adsorbed by the gel was higher using the Sephadex LH-20 when sera containing unconjugated bilirubin were tested. In case of sera containing also conjugated pigment, the bilirubin adsorbed by the two gels was the same at a Br/Alb molar ratio equal to or below 0.89; above this level the bilirubin adsorbed by the Sephadex LH-20 was higher. But with reference to the concentration of the unconjugated bilirubin the chloroform-recovered bilirubin was in greater amount than expected if unconjugated bilirubin were considered in the tested sera. The possible implication of the role played by the conjugated fraction is discussed in relation also to the clinical application of the gel filtration technique.


Water content and distribution in intrauterine growth-retarded newborn rats. Body composition of 54 normally grown and 29 intrauterine growth-retarded rat fetuses was determined. Half the fetuses in each group were analyzed whole while in the other half abdominal contents were separated from carcasses before analysis. No differences in body composition of whole fetuses, carcasses and abdominal contents were evident between the two groups. Although the validity of using the rat as animal model for human intrauterine growth retardation appears to be questionable, analysis of our results in light of previously published data raises important questions that deserve further investigation.


Effects of vaginal delivery and caesarian section on plasma renin activity and angiotensin II levels in human umbilical cord blood. High levels of angiotensin II were found in umbilical venous blood of babies delivered vaginally (40.3 pg-ml-1) and vaginally with epidural anaesthesia (66.8 pg-ml-1); low levels of angiotensin II were found in umbilical venous blood of babies delivered by Caesarian section (7.5 pg-ml-1) and in the peripheral blood of normal adults (7.92 pg-ml-1). There were no significant differences between these groups in the levels of plasma renin activity (PRA), although the mean values of PRA showed trends similar to those described for angiotensin II. It is suggested that the increase in PRA may account, in part, for the high levels in angiotensin II seen following vaginal delivery, but additional factors may also be involved.


Functional autonomic innervation of mammalian cardiac pacemaker during the perinatal period. Sinoatrial node pacemaker tissues from perinatal and adult rabbits, guinea pigs and rats were examined in vitro. Changes in spontaneous pacemaker rate produced by stimulation of intranodal vagal and sympathetic nerve endings, were taken as a measure of functional postganglionic innervation of the pacemaker. Results show marked species differences in the development of functional innervation of the cardiac pacemaker in the perinatal period.


Electrophoretic and biometric analysis of serum proteins of newborn babies. The average values and scatter of total protein and serum protein separated by paper electrophoresis of umbilical artery blood sampled at 130 normal deliveries, were determined. The distribution of the values was examined by means of a dot diagram. On the basis of the examination of an international standard preparation the reproducibility of the results is stated.


Glycerol metabolism in the pregnant and virgin rats. The parameters of in vivo glycerol kinetic curves were determined after a single injection of glycerol-1-14 C in pregnant and in virgin rats. The half-life of glycerol was found to be 9 min in pregnant rats and 17 min in virgin rats. There was no significant difference between the absolute turnover rates. The glycerol plasma space cannot be appreciated. The distribution volume expressed in milliliter of plasma appeared to be very high indicating that glycerol was not distributed only in the plasma. Glycerol was found to be a precursor of glucose and this conversion seemed to proceed more rapidly in the pregnant rat than in the virgin rat.


Separation and characterization of red blood cells from newborns and infants during the first trimenon of life by means of a dextran density gradient. Density distribution curves of erythrocytes and reticulocytes. Longitudinal studies on the density of erythrocytes and reticulocytes were performed by means of the isopyknic density gradient centrifugation in dextran on three mature newborns and six premature infants during the first 3 months of life. Striking changes of the density distribution curves of red blood cells took place during the first trimenon. Median density (D50) reached its maximum between the 7th and 28th day of life. D50 of reticulocytes was lower than that of the total red blood cell population and shows similar changes to the whole cell population during the first trimenon after birth. Based on the close relationship between D50 and the percentage of lightest cells, the evaluation of the percentage of the lightest cells was recommended as a simple test to study the changes in median density.


Developmental aspects of amino acid transport. Suggestion of switching-on and switching-off mechanisms. L-Lysine-14 C uptake was studied in vitro in intestinal segments of rats, in various ages, ranging between 15-day fetuses and adults. L-Lysine in 0.065 mM concentrations was accumulated against a chemical concentration gradient by processes which obeyed saturation kinetics. There appeared to be two peaks of lysine uptake values, one in the fetus, diminishing by the 17th fetal day and another on the 2nd postnatal day. Fetal transport of lysine was not inhibited by anaerobic conditions and was not Na+ dependent, in contrast with increasing O2 and Na+ dependency postnatally. A series of amino acids including representatives of the neutral, imino acid and dibasic groups, failed to inhibit lysine uptake, with the exception of L-arginine, which was also antagonized by L-lysine. These findings suggest that in the rat intestine L-lysine is transported by at least two mechanisms, one fetal, not requiring O2 and Na+, and another developing postnatally with a peak during the 2nd day, which is Na-dependent and requires aerobic conditions.


Agar ingestion combined with phototherapy in jaundiced newborn infants. 49 jaundiced, nonimmunized newborn infants with a birth weight of more than 2,000 g were given phototherapy with white light for more than 36 h. The average period of treatment was 61 h. 24 infants received 250 mg agar at the beginning of each meal at 3-hour intervals during phototherapy. 25 infants received phototherapy only. Serum bilirubin levels were decreased similarly in both groups. It is concluded that ingestion of agar does not supplement the effect of phototherapy.


Preceding pregnancy loss as an index of risk of stillbirth or neonatal death in the present pregnancy. Details of 9,311 singleton births born to multiparae in the first week of March 1958 were compared with 3,707 singleton cases of stillbirth and neonatal death delivered in the 3-month period March-May, with respect to the outcome of the preceding delivery. It was shown that the risk of death was raised some 66% if the preceding delivery had been an abortion and by a factor of nearly 3 if the preceding delivery had itself been a stillbirth or neonatal death (SBND). The increased risk was especially pronounced for macerated stillbirths. Analysis because of death revealed very significant increases in the incidence of congenital defects, asphyxia and 'no major lesion' when the preceding delivery had been either an abortion or an SBND. It was shown that, in general, a woman who had had a preceding SBND was over 3 times more likely to delivery by 35 weeks than on whose preceding delivery had ended in either an abortion or a survivor.


Epithelium and associated lymphocytes of developing human fetal appendix. Electron microscopic study. The epithelium and lamina propria of the human fetal appendix between 7.5 and 18 weeks of gestation was examined with electron microscopy. At 7.5 weeks the epithelium was composed of stratified columnar cells; scattered mesenchymal cells appeared in the lamina propria. By 13.5 weeks of gestation the epithelium was composed of simple columnar cells with a microvillous surface, goblet cells containing numerous mucinogen granules and argentaffin cells with elongated electron-dense granules. Also at 13.5 weeks of gestation some free lymphocytes were observed within the lamina propria as well as an occasional migrating lymphocyte within the epithelium between the columnar cells. At 18 weeks of gestation the simple columnar epithelial cells were tightly packed, the goblet cells were secretory and lymphocytes again appeared singly within the epithelium and lamina propria. No evidence of groups of lymphocytes or developing primary follicles within the lamina propria was observed in this study.


Estimation of "safe doses" in carcinogenic experiments. The statistical methodology for carcinogenic safety testing here developed has the following advantages: (1) Rather than making possibly unwarranted assumptions about a minimum slope in a dose response relationship, the present method represents an objective data based method of estimating "safe" doses. It is applicable to any specified permissible risk (of exceeding the spontaneous rate) and the latter must of course be specified by F.D.A. (2) Although the model which is used for the estimation of safe doses and their lower confidence points is parametric it comprises an adequately large number of parameters to allow for differences in the idiosyncracies of suspected carcinogens, host species and to a limited degree for variations in the experimental protocols. (3) The same computer program will cover the analysis of an experiments in which times to tumor have been recorded as well as experiments in which only tumor incidence rates have been recorded or mixtures of the two. "Better experimentation" is "rewarded" in that the lower confidence limits for the "safe doses" should be higher and more closely approach the true safe dose as the experimental effort increases. (4) The maximum likelihood estimation procedure is sophisticated and has asymptotic optimality properties. It utilizes the latest techniques of "convex programming" and the computer algorithm is straightforward and fast. The methodology proposed here also has the following shortcomings: (1) The model is (multi)parametric. However, it is of the form of the product model for age specific hazard rates which is now widely accepted. (2) Robustness studies on the effect of model breakdown on the estimated safe doses are as yet limited and should be followed up with more extensive studies. (3) Obviously no estimates of "safe doses" can be made if the spontaneous incidence rate is zero and the experimental dose levels have been chosen too small and no tumors have been observed. Similarly if the experimental doses are too small and the tumor incidence rates are all comparable with the spontaneous rate the estimation procedure is afflicted by extremely large errors. The situation improves slightly if the incidence for the highest dose level is higher than that of the lower dose levels which are all approximately equal.  In such situations more satisfactory experimental data are needed. Some general recommendations are as follows: (1) Whenever possible it is preferable to record times to tumor and not just incidence rates. However, for experiments of sufficiently long duration necropsies following the varying times of death will provide adequate information on the time dependence of tumor incidence. It may also be advisable to deliberately vary the times of sacrifice to two or three different times. (2) Other considerations being equal it is preferable to have a large number of dose levels rather than more animals per dose level...


Avoidance-modified generalised distributions and their application to studies of superparasitism. Given an underlying distribution for the number of encounters between parasites and a host this may be generalised by the distribution of the number of eggs per encounter. If eggs are already present, a parasite may avoid oviposition at subsequent encounters. A class of avoidance-modified generalised distributions is presented to model such situations. The case of a solitary (single egg-laying) parasite and a Poisson distribution for the number of encounters gives rise to a distribution of particular interest. Various properties of this distribution are derived and alternative estimation procedures investigated. Estimators based on the method of mean and zero frequency are shown to have the dual advantages over other estimators that have been proposed of simplicity of computation and increased efficiency. Over a wide range of parameter values there is virtually no loss of efficiency in comparison with maximum likelihood estimation.


Fitting a model to the growth of yeast colonies. When yeast cells reproduce, scars are left on the parent cell where the offspring has budded. Using a branching process model, it is possible to obtain the expectations of numbers of cells with 0, 1, 2, ... offspring. In this paper, theoretical results are tested against empirical data for three types of yeast cells. We examine the hypothesis that birth and death rates of cells with no previus offspring may differ from those of cells with one or more offspring. It is suggested that the oscillatory empricial results for the proportions of cells with 0, 1, 2, 3 and 4 offspring may be due to different mean budding times for these cells.


A general methodology for the analysis of experiments with repeated measurement of categorical data. This paper is concerned with the analysis of multivariate categorical data which are obtained from repeated measurement experiments. An expository discussion of pertinent hypotheses for such situations is given, and appropriate test statistics are developed through the application of weighted least squares methods. Special consideration is given to computational problems associated with the manipulation of large tables including the treatment of empty cells. Three applications of the methodology are provided.


The measurement of observer agreement for categorical data. This paper presents a general statistical methodology for the analysis of multivariate categorical data arising from observer reliability studies. The procedure essentially involves the construction of functions of the observed proportions which are directed at the extent to which the observers agree among themselves and the construction of test statistics for hypotheses involving these functions. Tests for interobserver bias are presented in terms of first-order marginal homogeneity and measures of interobserver agreement are developed as generalized kappa-type statistics. These procedures are illustrated with a clinical diagnosis example from the epidemiological literature.


Quantit analysis: a quantal assay refinement. A new tolerance distribution and inference procedure for quantal response assays is presented. This method is capable of fitting a wide variety of shapes of response curves. Specific cases of this new method include quantal response assay analyses based on the double exponential distribution, the logistic distribution (logit analysis), and the uniform distribution (linit analysis) in a limiting case. Computational techniques used to implement the likelihood procedures associated with this method are described. Comparisons are made for 22 sets of published data. These comparisons suggest that interval estimates of extreme dosages (e.g., ED95 and ED99) based on logit and probit analyses are, for the most part, either overly optimistic (too small) or overly pessimistic (too large). This interval estimation problem should be partially overcome by the added flexibility of the method introduced here.


Quadratic analyses of reciprocal crosses. Three different models, a two-way factorial model for familiarity, an orthogonalizing transform of this model to a diallel model, and a bio model more representative of the biological situation, are interrelated in terms of their components of variance and covariance. It is clarified that there are five components that can be reckoned with in the analysis of reciprocal crosses, including distinct maternal and paternal variances. Estimation of the components and tests of hypotheses concerning them are outlined for two types of mating designs with reciprocals. One deisgn involves a factorial mating design between two distinct sets of parents or parental lines and the other a diallel of all crosses from a single set of parents or parental lines. Both designs provide the same types of information and similar tests of hypotheses. At least some parts of the analyses corresponding to the factorial model are required to separate the maternal and paternal variances. A least squares partitioning of the sums of squares according to the diallel model, but with expectations expressed in terms of the bio model, provides most of the tests of hypotheses of interest. Worked examples are given.


Quick test for comparing two populations with bivariate data. A simple nonparametric test is given for testing the equality of two populations when the observations (X, Y) are bivariate and the alternative of interest is that the conditional c.d.f. of Y given X for one population dominates that for the other for every value of X. The procedure consists of fitting a curve through the combined set of data points and considering and conditional distribution of the number of points above the curve from one population, given certain information (depending on the design of the experiment), on the combined set of data. When used to compare two bivariate distributions, the test reduces to Fisher's exact test for 2 X 2 tables.


Goodness-of-fit tests for discrete data: a review and an application to a health impairment scale. We review the advantages and disadvantages of several goodness-of-fit tests which may be used with discrete data: the multinomial test, the likelihood ratio test, the X2 test, the two-stage X2 test and the discrete Kolmogorov-Smirnov test. Although the X2 test is the best known and most widely used of these tests, its use with small sample sizes is controversial. If one has data which fall into ordered categories, then the discrete Kolmogorov-Smirnov test is an exact test which uses the information from the ordering and can be used for small sample sizes. We illustrate these points with an example of several analyses of health impairment data.


Advantages of examining multicollinearities in regression analysis. In this paper a regression analysis is performed with data on spinal cord injuries in order to demonstrate the benefits of determining which, if any, multicollinearities are present in prediction data. Existing multicollinearities are shown to be useful both in determining characteristics of the sampled population as well as explaining possible erratic behavior of variable selection procedures. Latent root regression is performed on the data to illustrate one method of using biased regression techniques to incorporate knowledge of multicollinearities in developing prediction equations.


The analysis of the two-period repeated measurements crossover design with application to clinical trials. The two-period repeated measurements crossover design is often employed in clinical trials. This paper presents a unified approach for the statistical analysis of such a design that clarifies the testing procedures and assumptions employed under different conditions. It is shown how the data may be transformed so that it could be analyzed under the framework of a completely randomized repeated measurements design. Applications are given to a comparative bioavailability trial for attainment of steady state levels and to a clinical trial to compare the effects of two hypolipidemics.


Combination of a preference pattern with the triangle taste test. When dealing with the problem of obtaining information concerning consumer preferences for two competing brands of a product, one would like to assume that the consumers sampled can actually distinguish between the two brands. Often this is not the case and we have combined the standard triangle test of sensory perception with a preference test in order to obtain information concerning the consumers' ability to distinguish between the two brands. Assuming an underlying multinomial distribution, the maximum likelihood estimators (MLE) of the conditional probability that a person prefers brand A given that he can discriminate between the two products is then obtained. The estimator obtained is a ratio of linear combinations of observed multinomial proportions. The variability of this estimator is assessed by means of the jackknife statistic, the asymptotic variance of the MLE, and Monte Carlo studies. Application of the results obtained to taste test involving two brands of potato chips is discussed.


Further aspects of a Markovian sampling policy for water quality monitoring. In this paper, a Markov process is developed as a mathematical model to study the general problem of quality control monitoring. This approach was previously used by Arnold (1970) in development of sampling plans to study the water quality monitoring of streams. Arnold considered the expected sample size required for various sampling plans. Thre present authors have extended Arnold's work and derived equations for the variance of the sample size. In addition, approximation schemes are developed that could greatly enhance the implementation of this sampling procedure. Techniques are presented showing that an accurate approximation to the expected sample size can be obtained simply by the use of a hand calculator and equations developed within the paper. Moreover, accurate approximations for the variances of sample size can easily be obtained.


Random censoring and dendritic trees.

Random censoring and dendritic trees. The motivating problem is the estimation of the branching parameters of dendritic trees when some of the branches are cut. A primary element of this problem is the estimation of a bivariate discrete distribution when, because of partial censoring, some of the observations are incomplete. (This is similar to the problem of a multinomial cross-classification with supplemental partially cross-classified data.) Tests for randomness of censoring and estimators of parameters under random censoring are derived. The results on random censoring are applied in the dendritic tree context to obtain estimators for certain branching and cutting parameters. The testing and estimation procedures are applied to a set of dendritic tree data.


Velocity distributions of the streaming protoplasm in Nitella flexilis. Laser light is Doppler-shifted in frequency by the streaming endoplasm of living cells of Nitella flexilis. The frequency spectrum of the scattered light can be interpreted as the histogram of velocities within the organism, with the exception of the intense low-frequency portion of the spectrum. We demonstrate that the lowest-frequency component is the result of amplitude modulation of the scattered light by the array of chloroplasts in the cell. Measurement of the streaming endoplasm in a photobleached "window" region allows correction of the frequency distribution for the modulation component. The complete velocity histogram for the streaming endoplasm is calculated directly from the corrected frequency distribution. Measurements of vacuolar and endoplasmic motions show that the tonoplast, the membrane separating the vacuole and the endoplasm, seems to be flowing along with the endoplasm and vacuolar sap. Placing the cell in medium containing ATP in concentrations greater than 10(-3) M greatly increases the contribution of low velocities to the velocity histogram. Cytochalasin B at high dosages (10-50 mug/ml) does not noticably change the shape of the velocity histogram, while at low dosages (1 mug/ml) there is an increase in the contribution of low velocities to the velocity histogram. Colchicine in high concentrations (1%) has no observable effect on the velocity histogram.


Hydrogen bonding of adenine derivatives to tyrosine side chain. High resolution proton magnetic resonance measurements provide evidence for the formation of hydrogen-bonded complexes between 9-ethyladenine and p-cresol used as a model of tyrosine side chain in CDCl3. We have calculated the sum of the association constants corresponding to the three existing 1:1 complexes: K=6.3+/-0.15. By methylation of the amino group of adenine, we were able to calculate the ratio of the two strongest equilibrium constants K7/K1=1.6+/-0.3. Theoretical computations by the complete neglect of differential overlap (CNDO/2) method indicate that several hydrogen-bonded planar complexes can form between 9-methyladenine and phenol. The computed energy of the complexes with 6-dimethylamino adenine removes some ambiguity concerning the computed ratio of the association constants. Comparison of the calculated energies with free energies experimentally determined in organic solvent shows that despite the competition with CDCl3, which associates with both solute molecules, the preferential order of association is conserved. The small variations of charge density of adenine carbon atoms when complexed with phenol are in agreement with very small chemical shifts observed by 13C-nuclear magnetic resonance.


Evidence that nystatin may not form channels in thin lipid membranes. The mechanism of nystatin-induced conductance increases in planar bilayer membranes formed from lecithin/cholesterol (1:1) has been examined by spectral analysis of kinetic fluctuations. Lorentzian components of the power spectrum, characteristic of channel activity, were not observed for nystatin-doped membranes. Instead, the noise associated with this system (approximately 10(-27)A2/Hz) was in better agreement with that expected of a carrier mechanism involved in the transport of single charges in uncorrelated fashion. The implications of this result with respect to previous models of nystatin-induced conductance increases are discussed.


On the voltage-dependent action of tetrodotoxin. The use of the maximum rate-of-rise of the action potential (Vmax) as a measure of the sodium conductance in excitable membranes is invalid. In the case of membrane action potentials, Vmax depends on the total ionic current across the membrane; drugs or conditions that alter the potassium or leak conductances will also affect Vmax. Likewise, long-term depolarization of the membrane lessens the fraction of total ionic current that passes through the sodium channels by increasing potassium conductance and inactivating the sodium conductance, and thereby reduces the effect of Vmax of drugs that specifically block sodium channels. The resultant artifact, an apparent voltage-dependent potency of such drugs, is theoretically simulated for the effects of tetrodotoxin on the Hodgkin-Huxley squid axon.


Circular dichroism, optical rotatory dispersion, and absorption studies on the conformation of bovine rhodopsin iw situ and solubilized with detergent. Circular dichroism, optical rotatory dispersion and absorption of rhodopsin, the visual pigment of bovine rod outer segment membranes, were studied in situ and in membranes solubilized with various detergents. The alpha-helical content of the membrane protein is approximately 30%. The membrane protein possesses little beta-structure. Solubilization of the membrane by the detergents, Emulphogene BC-720 and cetyltrimethylammonium salts, results in loss of protein helical structure and perturbation of aromatic residues. These effects are not observed on digitonin solubilization. In regard to the structural stability of the membrane during bleaching, the following conclusions were reached: (1) Delocalized conformational changes of rhodopsin in situ involving secondary and/or tertiary structure are very unlikely. (2) Localized conformational changes of rhodopsin in situ involving secondary structure must be limited to the involvement of no more than three amino acid residues and localized conformational changes involving tertiary structure must be limited to very short segments of the protein chain containing, at the most, only a few aromatic residues. (3) Large changes in the interaction of lipid and protein moieties of the membrane are unlikely. (4) The detergents, Emulphogene, cetyltrimethylammonium salts, and digitonin, significantly decrease the conformational stability of rhodopsin as compared to the in situ conditions. The effect is smaller with digitonin. Evidence is presented against a proposed mechanism by which optical activity of the prosthetic group, retinal, is induced by resonance coupling of the transition dipoles of retinal and the lowest energy transitions of the aromatic groups of the apoprotein, opsin. A mechanism in which atropisomers of retinal are preferentially bound by opsin is consistent with the present results. The optical activity of the prosthetic group is markedly changed upon solubilization of the membrane by detergent. This change in optical activity is probably coupled to changes in conformation of the protein moiety induced by solubilization.


Quantitative model for the electric response of invertebrate and vertebrate photoreceptors. We propose that the same mechanism which leads to light-adaptation in invertebrate photoreceptors is responsible for the excitation of the receptor potential in vertebrates. Several qualitative and quantitative features of the vertebrate receptor response support this hypothesis.


The use of alpha-galactosidase and invertase in hollow fiber reactors. Invertase and alpha-galactosidase have been immobilized in hollow fiber cartridges with no detectable enzyme leakage and used for the hydrolysis of sucrose and raffinose, respectively. For both hollow fiber immobilized enzymes nearly complete substrate conversion is possible. Enzyme stabilities in polysulfonate hollow fibers which have been preconditioned with bovine albumin approach the stabilities of the free enzymes.


Effects of quantitative shock loadings on the constant recycle sludge concentration activated-sludge process. The reliability of the process of Ramanathau and Gandy (Biotechnical Bioeng., 13, 125 (1971) for the completely mixed activated-sludge process holding the recycle cell concentration, Xr, as a system constant with respect to step changes in hydraluic retention time was investigated. The experiments were run at initial dilution rates of 1/8, 1/6, 1/4, and 1/2 hr-1 treating a soft drink bottling wastewater. The influent substrate concentration was maintained at 1000 mg/liter chemical oxygen demand and the hydraulic recycle ratio at 0.3. The recycle sludge concentration was maintained at about 7000 mg/liter. It was found that the system could accomodate hydraulic shock loads up to 200% positive changes and down to 50% negative changes without disruption of the effluent quality. Shorter retention time of the range studied, from 2 to 8 hr, has the advantage of shorter response time with respect to the response of the concentration of biological solids in the reactor.


Characteristics of hydrocarbon uptake in cultures with two liquid phases. In hydrocarbon fermentation, the efficiency of hydrocarbon uptake by cells is one of the keys to the economical production of single-cell protein. This work is concerned with characterization of cultures with two liquid phases for understanding the hydrocarbon uptake process by cells. Batch cultivation of Candida lipolytica was carried out in shaking flasks and in a tower fermentor with motionless mixers. Microscopic observation and cell and hydrocarbon concentration distribution in batch cultivation showed that some cells are attached to the large oil drops and others are free from them. Interfacila tension between oil and water and Sauter mean drop size decreased as cultivation proceeded. On the basis of the experimental results, the process of hydrocarbon uptake by cells is discussed.


Colony formation in vitro by mouse blood monocytes. Mouse blood monocytes were induced to proliferate and form discrete colonies of mononuclear phagocytes in liquid culutre. The proliferation of these cells in vitro required a factor or factors present in medium conditioned by L cells. For this class of colony-forming cells, the value of Do to gamma irradiation in vitro was 195 rads.


Cell size and plamitate-1-14c turnover of rabbit marrow fat. Adipose tissue is a major component of normal rabbit marrow. Morphological considerations suggest an active role for this tissue in hematopoiesis. This hypothesis was tested by injecting 50 micronCi of palmitate-1-14C intravenously into fed, hematologically normal New Zealand rabbits. The animals were sacrificed 24 hr later and the femoral marrow removed. Samples of subcutaneous and perinephric fat were taken for comparison. The fat cells were isolated by the Rodbell method and the diameters measured. Incorporation of the 14C-palmitate in the triglyceride fraction was determined and the composition of the fatty acids was measured by gas chromatography. The mean diameter of the marrow fat cell was 46 micronm (mean cell volume 55 pl); the mean diameter of the perinephric fat cell 70 micronm (mean cell volume 200 pl). 14C-Palmitate turnover per gram triglyceride was some fivefold greater in the marrow fat; however, when expressed on a cell basis, the turnover for the marrow and perinephric fat cell was similar. The marrow fat contained a higher concentration of unsaturated fatty acids. These findings suggest that there is greater lipolysis and lesser storage in the marrow fat than in the perinephric.


Carriers with excessively low factor VIII procoagulant activity (VIII AHF): a study of two unrelated families with mild hemophilia A. Two unrelated families are described with mild hemophilia A in whom six obligate carriers had unusually low VIII AHF levels. In each family, successive generations of males were affected with hemophilia A as determined by low VIII AHF in the presence of normal VIII AGN and VIII VWF levels. In the first family, two of five obligate carriers had low VIII AHF levels associated with clinical bleeding and one other had a history of bleeding. While receiving oral contraceptives, one of these two carriers was found to have a normal VIII AHF level. In the second family, four cousins below age 10 who were obligate carriers had significantly low VIII AHF levels, while a paternal aunt and paternal grandmother who were also obligate carriers had VIII AHF levels within the normal range. Hemorrhagic diathesis in multiple obligate carriers in these families is not readily explained by the Lyon hypothesis, and suggests that these families may be exmaples of an unusual allelic form of hemophilia A or that they may be transmitting several independent genes affecting VIII AHF levels. Our experience suggests that VIII AHF levels should be determined on all obligate or possible carriers prior to surgery to identify those individuals at risk for postoperative bleeding. Furthermore, it is suggested that hormonal therapy might be effective in the management of carriers with low levels of VIII AHF and clinical bleeding.


Demonstration and mode of action of an inhibitor for activated Hageman factor (factor XIIa) of the intrinsic blood coagulation pathway from Schistosoma mansoni. An anticoagulant activity from adult Schistosoma mansoni whole worm homogenate is described. The inhibitor appears to be specific for the contact activation step of the intrinsic pathway. Experiments with both human and mouse plasmas have defined the specificity of the inhibitor as follows: (1) It lengthens the partial thromboplastin time of normal plasma. (2) It has no effect on the prothombin time and Russell's viper venom time of normal plasma. (3) Preactivation of normal plasma by a contact activator such as Celite eliminates essentially all inhibitory activity. (4) The inhibitor appears to be heat stable and can be precipitated by centrifugation above 27,000 g. (5) The inhibitor has no effect on the activation of factor XII by Celite. (6) The activation of factor XI by factor XIIa, however, is inhibited by the schistosomal inhibitor. The above data are consistent with the view that S. mansoni adults possess an anticoagulant that is capable of specifically inhibiting the conversion of factor XI to factor XIa by factor XIIa.


Significance of the intact polypeptide chains of human fibrinogen in ADP-induced platelet aggregation. The presence of human fibrinogen in suspensions of washed human platelets is a requirement for ADP-induced platelet aggregation. Digestion of fibrinogen with plasmin destroys this function of the protein. The high solubility fraction of Kabi fibrinogen, fragment X (stage 1) and framgent X (stage 2), are two, eight, and ten times, respectively, less potent in promoting ADP-induced platelet aggregation, as compared with intact fibrinogen. Fragments Y and D and the mixture of reduced and carboxymethylated chains of human fibrinogen do not support ADP-induced platelet aggregation at all. SDS polyacrylamide gel electrophoresis of nonreduced and reduced fibrinogen and its derivatives indicates that the intact fibrinogen molecule is essential for ADP-induced platelet aggregation. It is suggested that the carboxy-terminal part of the Aalpha chain and possibly also the amino-terminal part of the Bbeta chain are required for the platelet aggregation-promoting function of fibrinogen.


Effect of aspirin on thrombogenesis and on production of experimental aortic valvular Streptococcus viridans endocarditis in rabbits. Because thrombus formation at the site of endothelial injury has been thought to be a critical step in the pathogenesis of bacterial endocarditis, the effect of aspirin on experimental valvular thrombosis and bacterial endocarditis in rabbits was evaluated. Aortic valvular injury and thrombosis were induced in aspirin-treated and control rabbits with intracardiac catheters. A subsequent inoculation of Streptococcus viridans resulted in the development of infective endocarditis. Rabbits were sacrificed as early as 6 hr, and the effectiveness of aspirin was determined by the weight of the sterile vegetations and the quantitation of bacteria in the thrombotic vegetation. Aspirin, in levels in excess of 50 mg/dl did not attenuate the evolution of infective endocarditis, since the formation of sterile thrombotic vegetation and bacterial endocarditis in aspirin-treated rabbits was similar to those in controls.


Circulation of donor lymphocytes after blood transfusion in man. Atypical lymphocytes (ATL) appear in the circulation of a large proportion of patients during the first week following a blood transfusion. In order to determine the source of these ATL, cytogenetic studies were performed on the peripheral blood leukocytes of ten adult patients who received fresh blood from donors of the opposite sex. Nine of the ten patients had spontaneously dividing mononuclear cells of the recipient or host karyotype circulating during the latter part of the first week after transfusion. In two patients, the spontaneously dividing cells were of donor as well as of host origin. Six patients had circulating phyothemagglutinin-responsive lymphocytes of the donor karyotype noted from 1-7 days after transfusion. These findings lend support to our hypothesis that the increase in circulating atypical lymphocytes seen 1 wk after transfusion represents the counterpart in vivo of the in vitro mixed leukocyte reaction. The dividing donor cells may represent a subclinical graft-versus-host reaction.


Erythrocyte membrane plaques from rats with magnesium deficiency. This study investigated the anemia of dietary magnesium deficiency in inbred Fisher white rats using freeze-fracture electron microscopy. The plasma membranes of erythrocytes from animals receiving two different magnesium-deficient and control diets were observed at weekly or biweekly intervals for 6 wk. The earliest changes were small plaques on the external surface (ES) and fracture face (PF) of erythrocyte plasma membranes, which occurred after 2 wk of either magnesium-deficient diet. These plaques persisted and increased in size with progressive magnesium deficiency. When fully developed, the plaques consisted of round or oval elevations approximately 30-50 nm in diameter outlined by a narrow raised border. The surface of the plaques was smooth and devoid of intramembranous particles. Incubation of erythrocytes from magnesium-deficient rats in a physiologic solution containing 2 meq/liter magnesium for 1 hr at 37degrees C did not alter the appearance of the plaques. Erythrocytes from control rats, obtained during the same time periods, showed no plaques. Thus, a deficiency of magnesium in rats altered erythrocyte membrane structure.


Relative and absolute numbers of E- and EAC-rosette-forming cells in lymphoid organs. The percentage of lymphocytes of the blood and lymphoid organs of young pigs were determined by forming rosettes with pretreated sheep red blood cells (AET-rosettes) or with sheep red blood cells coated with antibody and complement (EAC-rosettes). The mean figures for AET-rosettes were 70% for the blood, 93% for the thymus, 78% for lymph-nodes and 68% for the spleen. For EAC-rosettes the numbers were 16%, 0%, 16% and 19% respectively. The absolute numbers of rosette-forming lymphocytes were estimated on the basis of previously determined numbers of lymphoid cells in young pigs. Even by excluding the data for the thymus, there would be about three times more AET-rosette forming lymphocytes than EAC-rosette forming cells in the whole animal. About 11% of all lymphocytes did not show any of these markers. Data on the numbers of "B" and "T" lymphocytes in human lymphoid organs from the literature were summarized and compared to those found in pigs and thus, the absolute numbers for a young adult man estimated.


Mycoplasma pneumoniae infection complicated by paroxysmal cold hemoglobinuria with anti-P specificity of biphasic hemolysin. A 7-year-old boy with Mycoplasma Pneumoniae infection complicated by transitory paroxysmal cold haemoglobinuria (PCH) is described. The Donath-Landsteiner-antibody exhibited anti-P specificity; hemolytic activity was partially inhibited against papainized erythrocytes at 0 degrees C incubation temperature and increased from 8 degrees C upwards. The association of Mycoplasma pneumoniae infection with PCH has been described 4 times only and in one instance where specificity was stated it was anti-I.


Platelet function, blood coagulation and fibrinolysis in Behcet's syndrome. In 10 patients with Behcet's syndrome, various parameters of platelet function, blood coagulation and fibrinolysis were studied. With varying frequency the following abnormalities were found: increased retention of platelets in glass bead column, reduced platelet aggregation to low concentrations of adenosine diphosphate, elevated plasma levels of fibrinogen concentration and factor VIII activity, increased plasma antiheparin activity and impairment of fibrinolytic activity. The above abnormalities were found long after the last thrombotic episode and were more frequent in patients with a history of thrombophlebitis. It is suggested that certain hemostatic abnormalities accompany or form part of Behcet's syndrome and that they are related to the thrombotic complications characteristic of this syndrome.


The utilization of a new strength citrate anticoagulant during centrifugal plateletpheresis. II. Assessment of in vitro platelet function. Platelets contained in concentrates prepared by intermittent flow centrifugation utilizing 5.0 g total ionized citrate, function normally as assessed by tests for adhesion, spreading, and ADP and collagen-induced aggregation. The ability of platelets to adhere and to aggregate following incubation with ADP is significantly reduced, in samples taken immediately after the collection procedure. Total platelet collection and platelet collection efficiency, is not compromised by use of the above citrate concentration.


Acute toxicities of selected heavy metals to the softshell clam, Mya arenaria. Static acute toxicity bioassays with adult softshell clams and salts of copper, cadmium, zinc, lead, manganese, and nickel were conducted at 30 0/00 salinity and 22 degrees C. Concentrations fatal to 50% in 168 hours, in mg/l (ppm) metal added at start, were 0.035 for Cu, 0.150 for Cd, 1.55 for Zn, 8.80 for Pb, 300.0 for Mn, and less than 50.0 for Ni. Additional tests were conducted with Zn2+ and Cu2+ at 30 0/00 during fall (17.5 degrees C) and winter (4 degrees C); clams displayed increasing survival with decreasing temperature. For Cu, LC-50 (336 hr) values at 17 degrees C and 4 degrees C were 0.086 and greater than 3.00 mg/l, respectively; for Zn these were 2.65 and greater than 25.0, respectively.


Protective action of selenium against mercury in northern creek chubs. We found selenium dixoide an effective antagonist to the toxic effects of mercuric chloride in the northern creek chub (Semotilus atromaculatus). Selenium pretreatment increased the whole body accumulation of mercury at lower environmental Hg concentrations. It decreased mercury accumulation in survivors at higher, acutely toxic concentrations.


Chlorinated hydrocarbon insecticide residues in Tennessee honey and beeswax. Several samples of commercial grade honey collected from different parts of tennessee during the summer of 1973 were analyzed for chlorinated hydrocarbon insecticide (CHI) residues. A "Modified Mill's Procedure" was used to cleanup the samples prior to gas chromatographic analysis using electron capture (EC) detection. The presence of CHI residues was confirmed by analysis on three different columns of widely varying polarity. Most of the samples contained CHI residues at 0.01-0.30 parts per billion (ppb) level. Beeswax produced during the same season contained several times higher levels of the residue than the honey samples. Recoveries of CHI residues varied from 81-95 percent by the procedure employed.


Deposition of heavy chlorinated hydrocarbons from the atmosphere. A sensitive method for the measurement of deposition fluxes of volatile and aerosol chlorinated hydrocarbons (CHC) is presented. Fluxes of CHC ranging from 10(-6) to 10(-9) g m-2 day-1 for the 54% chlorine PCB mixture were measured at La Jolla, California (average: 3 x10(-7) g m-2 day-1) during February, 1972. PCB fluxes are correlated with the component of the wind trajectory in the direction of Los Angeles weighted by the reciprocal of the wind speed. Chlorinated pesticide fluxes (DDT and dieldrin) are correlated with winds from agricultural areas inland from the measuring site.


Effect of nickel sulphate on male rats.

Effect of nickel sulphate on male rats. Cytopathological and histopathological changes in skin, liver, kidney and testis of rats due to nickel sulphate have been studied experimentally. Skin of nickel sulphate painted rats showed hyperkeratinization, vacuolization, hydropic degeneration of basal layer and atrophy of epidermis. The testis showed degeneration and oedema of seminiferous tubules, while liver showed areas of focal necrosis, congestion and dilatation of sinusoids.


The organization of successful participative management in a health sciences library. The University of Washington Health Sciences Library, Seattle, and its participative management process are described in detail. The evolution of the management system is reviewed by interrelating the various phases of the library's growth, its service complexities, and its communication needs. Staff development results of this participative management mode are discussed. Reference is made to the use of group dynamics concepts. The current organizational design, which integrates the participative subunit with the simple line management structure, is considered effective by both the library staff and its director.


Coordinators for health science libraries in the Midwest Health Science Library Network. In the summer of 1973 one resource library in each of the six states of the Midwest Health Science Library Network received funding from the National Library of Medicine to hire a coordinator for health science libraries. The development of the role of coordinator is examined and evaluated. The coordinators have proved valuable in the areas of consortium formation, basic unit development, communication facilitation, and program initiation. The function of the coordinators in the extensive planning effort now being undertaken by the network and the future need for the coordinator positions are discussed.


Centralized automated cataloging of health science materials in the MLC/SUNY/OCLC shared cataloging service. Since February 1976, The Medical Library Center of New York, with the assistance of the SUNY/OCLC Network, has offered, on a subscription basis, a centralized automated cataloging service to health science libraries in the greater metropolitan New York area. By using workforms and prints of OCLC record (amended by the subscribing participants), technical services personnel at the center have fed cataloging data, via a CRT terminal, into the OCLC system, which provides (1) catalog cards, received in computer filing order; (2) book card, spine, and pocket labels; (3) accessions lists; and (4) data for eventual production of book catalogs and union catalogs. The experience of the center in the development, implementation, operation, and budgeting of its shared cataloging service is discussed.


The effect of chronic exposure to tobacco smoke on the antibacterial defenses of the lung. To evaluate the effects of cigarette smoking on the host defenses of the lung, male CD rats were exposed to fresh whole smoke for up to 60 consecutive days. Intrapulmonary deposition of smoke and animal exposure levels, quantified with decachlorobiphenyl and other smoke tracers, indicated a daily cigarette exposure equivalent to approximately a pack and a half per day in man. Pulmonary alveolar macrophage function in situ was quantified by the inactivation of an aerosolized challenge of Staphylococcus aureus six hours after inoculation. Controls (n=120) inactivated 88.8+/-0.64% of the staphylococci. Exposure to whole smoke did not impair intrapulmonary antistaphylococcal defenses, with inactivation rates of 89.8+/-0.97% (n=49) and 89.1+/-0.46% (n=74) at 30 and 60 days, respectively. Inactivation distribution frequency analysis in controls revealed that 7% of animals had inactivation values greater than two standard deviations from the mean. With prolonged exposure mean with less skewing towards the abnormal. Alveolar macrophages harvested from smoked animals were comparable in viability and in vitro antistaphylococcal activity to controls, appeared to be metabolically activated and had specific stereologic ultrastructural alterations. These studies indicate that chronic exposure to tobacco smoke does not impair, and in fact may stimulate, the host defenses of the lung, as evaluated by in vivo and in vitro pulmonary alveolar macrophage function.


[Effects of nitrogen dioxide on alveolar macrophages surviving in the gas phase. A new experimental model for the study of in vitro cytotoxicity of toxic gases (author's transl)]. Alveolar macrophages surviving in the gas phase were exposed to concentrations of NO2 (0.1, 1 and 2 ppm) for 30 minutes. The exposed cells showed morphological changes, a decrease in bactericidal activity and a reduction of the ATP content. The severity of these changes is related to NO2 concentration. This new experimental model offers new possibilities for studies on the cytotoxicity of gases.


Estimating the satisfaction of information users. Several factors influence the responses of users of information retrieval systems who are asked to evaluate the service provided by a system. To realistically estimate user satisfaction, different aspects of the search outcome should be explored. This paper analyzes the data obtained in a follow-up questionnaire sent to a sample of MEDLINE users who had requested searches at seven information centers within a two-month period. The questionnaire requested the user's appraisal of the value of the search and of the usefulness of the retrieved references. A comparison of different items of the questionnaire revealed some contradictions between an overall appraisal of the service and more specific responses on the outcome of the search. Caution is advised in inferring the satisfaction of information needs from the users' evaluation of an information retrieval system.


[Comparative study of bronchial mucins isolated from the sputum of patients suffering from cystic fibrosis or other chronic bronchial diseases (author's transl)]. Most of the mucins isolated from the sputum of patients suffering from cystic fibrosis are acidic. Acidic mucins from a child suffering from cystic fibrosis were degraded by alkali treatment. Analysis of the degradation products demonstrated the wide heterogeneity of the carbohydrate chains linked to the mucin polypeptide moiety. However, this heterogeneity of carbohydrate chains is probably not restricted to mucins from patients with cystic fibrosis. Bronchial mucins were prepared either from the sputum of different patients belonging to blood group O or B and suffering from cystic fibrosis, chronic bronchitis and other chronic bronchial diseases, or from bronchial washings performed in macroscopically healthy area of the bronchial tree of subjects belonging to blood group O. The chemical composition of each mucin fraction was established and an average carbohydrate chain length was estimated. Acidic mucins isolated from the sputum of two children suffering from cystic fibrosis were more sulfated than sialylated and their average carbohydrate chain length was relatively large. These characters were not specific for cystic fibrosis since they were also found in acidic mucins of two children suffering from other bronchial diseases. Most of the acidic mucins isolated from the sputum of three adults suffering from chronic bronchitis were more sialylated than sulfated and had a relatively short average carbohydrate chain length. The sputum of these patients also contained a variable proportion of neural or weakly acidic mucins. The mucins isolated from bronchial washings performed in macroscopically healthy areas of the bronchial tree were acidic molecules whose acidic characteristics and average carbohydrate chain length were about the same as for acidic mucins from patients with chronic bronchitis.


COCTAILS: automated interlibrary loan statistics at health sciences library, SUNYAB. An automated interlibrary loan statistics compilation system used at the Health Sciences Library at the State University of New York at Buffalo is described. The system provides standardized input to produce various statistical reports as well as additional information for collection development and improved user service.


Excerpta Medica abstracting journals: a case study of costs to medical school libraries. A cost comparison study was made of Excerpta Medica's abstracting journals, based upon actual costs to a library. Unit costs were determined for six sections of EM as compared with six corresponding abstract journals. On average, EM sections were found to be 138% more costly than corresponding abstract journals. The effects of splitting of EM journal titles were also analyzed. This practice increases the price of a total subscription to EM and makes comprehensive information retrieval more difficult. A survey of medical school librarians as users of EM points to dissatisfaction with its increasing price, particularly when it results from title splitting.


A college course for nurses on the utilization of library resources. Library user instruction has been a no-man's-land between librarian and educator. Each assumes that the other has offered the student some necessary preparation before his assignments requiring library consultation. Too often, this is not the case. Reference librarians at the Duke University Medical Center Library are currently offering a ten-week, 1 1/2-hour credit library instruction course for nursing baccalaureate candidates. "Introduction to Library Resources in the Nursing Field" is designed not so much to orient students to a specific library facility, but rather, to provide them with background material on library organization and services and to familiarize them with basic bibliographic tools relevant to the nursing profession. Lectures are supplemented through the use of homework questions, bibliographies, handouts, in-class exercises, and on-line demonstrations. Very positive feedback from both students and faculty has attested to the value of such a course in the nursing curriculum and has resulted in its being offered four times to date. The library staff is exploring methods for offering additional library instruction not only to nursing students but to other user groups in Duke's medical complex.


New library buildings: the Houston Academy of Medicine--Texas Medical Center Library. A historical account is given of the Houston Academy of Medicine--Texas Medical Center Library within its Texas Medical Center setting in Houston, Texas. Outlined are planning, financing, and construction of the new library, which consists in part of new building and in part of renovated interiors of an old building originally completed in 1954. A concise picture is given of the new library's interiors, showing its functional success for users and employees alike. An architectural summary is appended showing gross and net footages, source of funds, costs and capacities.


New library buildings: the Augustus Long Health Sciences Library of the College of Physicians and Surgeons, Columbia University. A brief historical sketch, tracing the development of the College of Physicians and Surgeons and its library from the Royal charter date of 1754, is presented. Little is recorded of library development prior to 1928, when the departmental libraries were united with the Reference Library to form the core of the present library. The planning processes for the new library are discussed, noting the involvement of the entire library staff. Four floors in a twenty-story tower are devoted to library services, and each floor is described. Several of the major compromises made during the planning process are documented. Photographs and floor plans of two of the floors help illustrate the text.


Fevers of hot climates.

Fevers of hot climates. The febrile patient from the tropics is generally, but not invariably, suffering from an infection which need not necessarily be exotic although this possibility must be excluded. In acute infections malaria must always be considered, as its diagnosis and treatment are easy and effective. A positive diagnosis of an exotic infection can only be made if a history of long or short residence in a tropical or subtropical area is obtained. In most patients a diagnosis is readily established.


Prostaglandin F2alpha reduces the algesic effect of bradykinin by antagonizing the pain enhancing action of endogenously released prostaglandin E. 1 The isolated perfused ear of the rabbit connected to the body only by its nerve, was used to investigate the influence of prostaglandin F2alpha on the algesic effect of bradykinin and acetylcholine. 2 Bradykinin and acetylcholine, following intra-arterial injection into the isolated perfused ear elicited a dose-related reflex fall in blood pressure due to stimulation of paravascular pain receptors (= algesic effect). 3 Infusion of prostaglandin F2alpha (0.1 to 1 ng/ml) into the rabbit ear reduced the algesic effect of bradykinin but not that of acetylcholine. 4 The onset of the reflex fall in blood pressure by bradykinin but not that by acetylcholine was delayed by infusion of prostaglandin F2alpha into the ear. 5 Infusion of prostaglandin E1 into the rabbit ear led to an enhancement of the algesic effect of bradykinin and acetylcholine. Enhancement of both effects was abolished by infusion of prostaglandin F2alpha. 6 During inhibition of the endogenous synthesis of prostaglandins (mainly E-type) by indomethacin, a low concentration of prostaglandin F2alpha no longer reduced the algesic effect of bradykinin. However, a high concentration of F2alpha continued to enhance the effect of bradykinin and acetylcholine. 7 Prostaglandin F2alpha influenced neither the brief reduction in venous outflow produced by bradykinin nor the brief increase in venous outflow caused by acetylcholine. 8 The results suggest that prostaglandin F2alpha does not directly reduce the effect of bradykinin but inhibits the enhancement of its algesic effect produced by prostaglandin E that is released endogenously by bradykinin. That the algesic effect of acetylcholine is not reduced by prostaglandin F2alpha is in keeping with its releasing very little endogenous prostaglandin E.


Brain and vascular monoamine oxidase activity in the deoxycorticosterone-salt hypertensive rat. 1 The role of monoamine oxidase (MAO) in the maintenance of deoxycorticosterone-sodium chloride (DOCA-salt) hypertension was investigated by assaying the MAO activity both in central as well as peripheral blood vessels and in brain tissue. 2 The results suggest that the activity of MAO in the DOCA-salt hypertensive rat is similar to the activity present in the normotensive rat.


The specificity of oestrogen receptor in brain, pituitary and uterus. 1 The specificity of oestrogen receptor in rat brain regions (hypothalamus, amygdala and cortex), pituitary and uterus was studied by measurement of the inhibition of 17beta-oestradiol high affinity binding in cytosol, in the presence of unlabelled putative inhibitors of binding. 2 Binding was inhibited only by those of the compounds tested that possessed oestrogen agonist or antagonist activity. The affinities were estimated and the ranking order of the compounds was the same in all tissue sources of cytosol and corresponded to the ranking order of agonist or antagonist potency. 3 There were some significant differences between some of the estimated affinities in different tissues, these being seen most commonly between pituitary and hypothalamus on one hand and uterus on the other. 4 The possibility of heterogeneity of oestrogen receptor is discussed.


Effects of antidepressant drugs on noradrenaline accumulation and contractile responses in the rat anococcygeus muscle. 1 The effect of a series of antidepressant drugs on noradrenaline accumulation was studied in the isolated anococcygeus muscle of the rat. 2 The most potent inhibitors of noradrenaline accumulation were nortriptyline, desipramine and protriptyline. Opipramol, trimipramine and iprindole were active only in high concentrations. 3 Contractions of the anococcygeus muscle produced by noradrenaline were strongly potentiated by nortriptyline, desipramine and protriptyline. Other uptake inhibitors were less active in potentiating the noradrenaline response. 4 Nortriptyline, in concentrations that potentiated the action of noradrenaline, reduced or abolished the response to tyramine.