Techniques of infrared thermography.

Techniques of infrared thermography. Considering the main objectives of thermographic techniques and the particular characteristics of passive infrared detection, the essential arguments in favor of single detector scanners have been pointed out after a brief discussion of the detection and image pickup methods when quantitative and precise (0.1 degrees C) information on the temperature distribution near ambient temperature must be obtained. Single detector scanners can have sufficiently good performance to reach the objectives of precise thermography, but their real technical limits must be well-known in practice; consequently, the thermograph performance must be clearly defined by means of the modulation transfer function, the intrinsic thermal resolution and the response uniformity, as we indicate in the second part of this paper.


Immunoadjuvant activities of synthetic N-acetyl-muramyl-peptides or -amino acids. A variety of N-acetylmuramyl-peptides (or -amino acids) were prepared by condensation of benzyl N-acetyl-4, 6-O-brnzylidene-alpha-muramide with various peptide (or amino acid) benzyl esters by the dicyclohexylcarbodiimide--N-hydroxysuccinimide or ethylchlorocarbonate--N-methylmorpholine method and removal of the protecting groups by hyderogenolysis. N-Acetylmuramyl-L-alanyl-D-isoglutamine was identified as the minimum structural entity essential for the immunoadjuvant activities characteristic of bacterial cell walls. Consequently N-acetylmuramyl-L-alanine was not adjuvant active. The tetrapeptide portion of adjuvant-active N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-lysyl-D-alanine proved to be inert, at least in induction of delayed-type hypersensitivity. The possible adjuvant activities of various analogues or diastereomers of the above N-acetylmuramyl-dipeptide and related compounds were studied. N-Acetylmuramyl-L-alanyl-D-glutamic acid exhibited weak, but definite adjuvancy, but none of the others, including N-acetylmuramyl-L-alanyl-L-isoglutamine, N-acetylmuramyl-L-alanyl-D-glutamine and N-acetylmuramyl-L-alanyl-D-isoasparagine, had any adjuvant activity. This clearly indicated the importance of the configuration of the glutamic acid residue or its amides, i.e. the presence of the D-isoglutamine residue in the N-acetylmuramyl-dipeptide, for manifestation of adjuvant activities in stimulation of both antibody-mediated and cell-mediated immune responses. Neither N-acetylmuramyl-D-isoglutamine nor N-acetylmuramyl-D-alanine had any adjuvancy.


Subacute sclerosing panencephalitis (SSPE): isolation of a defective variant of measles virus from brain obtained at autopsy. A cytopathic agent causing formation of syncytial giant cells was isolated by co-cultivation of human embryonic lung cells with brain cells obtained at autopsy from a patient with subacute sclerosing panencephalitis. Measles specific intracellular immunofluorescence was detected in syncytial giant cells developed by the agent. Paramyxovirus-like nucleocapsids were observed by electron microscopy in nuclei of the syncytial giant cells. Measles specific immunofluorescence was also detected on the surface of unfixed syncytial giant cells. However, the synycytial giant cells did not produce either virions or hemogglutinin, and did not show hemadsorption of African green monkey red cells. Hence, the newly isolated agent seems to be a defective variant of measles virus, and was designated as the SSPE-"BIKEN" strain.


Immunoadjuvant activities of cell walls and their water-soluble fractions prepared from various gram-positive bacteria. The cell walls from all 21 species of gram-positive bacteria examined, except lysozyme-susceptible Micrococcus lysodeikticus (NCTC 2665) and lysozyme-resistant Staphylococcus epidermidis (ATCC 155), were found to be definitely adjuvant-active in both stimulation of increased serum antibody levels and induction of delayed-type hypersensitivity to ovalbumin when administered to guinea-pigs as water-in-oil emulsions. Using various cell wall lytic enzymes, the immunoadjuvant principles were solubilized with full retention of the adjuvant activities from walls of Staphylococcus aureus (Copenhagen), Streptococcus pyogens (group A, type 6; S43/100), Streptococcus salivarius (IFO 3350), Streptococcus faecalis (IFO 12580), Streptococcus mutans (BHT), Lactobacillus plantarum (ATCC 8014), Bacillus megaterium (IFO 12068), Corynebacterium diphtheriae (Park-Williams No. 8), Mycobacterium smegmatis, and Actinomyces viscous (ATCC 15987). Evidence was obtained that the non-peptidoglycan portion of the cell walls is not essential for manifestation of immunoadjuvancy.


Immunoadjuvant activities of peptidoglycan subunits from the cell walls of Staphyloccus aureus and Lactobacillus plantarum. Attempts were made to isolate and identify the unit chemical structure essential for manifestation of the immunoadjuvant activities characteristic of bacterial cell walls. The N-acetylmuramyl-peptide subunit monomers, Nalpha-(N-acetylmuramyl-L-alanyl-D-isoglutaminyl)-Nepsilon-(glycylglycyl)-L-lysyl-D-alanine from the cell walls of Staphylococcus aureus (FDA 209P) and N-acetylmuramyl-L-alanyl-D-isoglutaminyl-meso-diaminopimelic acid and/or N-acetylmuramyl-L-alanyl-D-isoglutaminyl-meso-diaminopimelyl-D-alanine from those of Lactobacillus plantarum (ATCC 8014), were shown to be unit chemical entities with definite adjuvant activity both in stimulation of antibody production and in induction of delayed-type hypersensitivity to ovalbumin when administered to guinea-pigs as water-in-oil emulsions.


Gene action in fish of tetraploid origin. II. Cellular and biochemical parameters in clupeoid and salmonoid fish. By use of cell size, protein and hemoglobin content, and enzyme activities as markers, it becomes apparent that in the course of evolution the gene expression of anciently tetraploid fish of the order Ostariophysi was diploidized, but no such regulatory mechanism has evolved in the phylogenetically tetraploid species of the order Isospondyli. This finding is discussed in terms of possible selective neutrality of tetraploid expression and the phylogenetic age of Isospondyli.


Polymorphism and linkage of glutathione reductase in Mus musculus. Using an electrophoretic variant for glutathione reductase, we have been able to map the structural gene for this enzyme to chromosome 8 in Mus musculus.


Study of a corn (Zea mays L.) mutant (blue fluorescent-1) which accumulates anthranilic acid and its beta-glucoside. A corn (Zea mays L.) mutant, blue fluorescent-1 (bf), is described that shows ultraviolet light induced blue fluorescence in young seedling leaves if homozygous for the mutant gene, and in anthers if either homozygous or heterozygous. The blue fluorescent compounds were extracted with acetone and separated by paper chromatography. Anthranilic acid was present and the beta-glucoside was also identified by paper chromatography and beta-glucosdase and acid treatment. A third major fluorescent compound was not identified, but it was convertible to anthranilic acid by acid treatment. Anthranilate synthetase from mutant plants was 3-40 times more active and was also more resistant to feedback inhibition by tryptophan than was the enzyme from normal plants. The high activity and feedback resistance would both lead to anthranilate accumulation. Anthranilate-phosphoribosylpyrophosphate phosphoribosyltransferase (PR transferase), the enzyme which usually utilizes anthranilate in the tryptophan pathway, was inhibited by the beta-glucoside of anthranilic acid in a noncompetitive manner and showed very little activity in the mutant plant extract. This inhibition of the enzyme which utilizes anthranilate would also lead to accumulation. Apparently the oversynthesis of anthranilate leads to the formation of the beta-glucoside, which inhibits anthranilate utilization. The fluorescent compounds are absent in seed, but form on germination. The levels decrease with age after 35 days postgermination, but are still present in leaves during grain filling.


Esterase. XXI. Es-9, a possibly new polymorphic esterase in Mus musculus genetically linked to es-2. A so far undescribed gene controlling zone III esterases has been detected by means of disc gel electrophoresis of kidney homogenates from the two inbred mice strains NMRI and SK/Cam. The gene is tentatively designated Es-9, and the two codominant alleles are designated Es-9a and Es-9b. Es-9 esterases are present in many tissues, but, unlike the other zone III esterase (controlled by Es-5), are not found in the serum. Close linkage with the Es-2 gene leads us to map the Es-9 gene on chromosome 8.


Molecular and genetic structure of multiple hemoglobins in the eelpout, Zoarces vivparus L. Six major hemoglobin components are present in the teleostean fish Zoarces viviparus L. Biochemical characterization has led to a molecular model for the polypeptide chain composition of the individual hemoglobins. Only three different chains are involved. They are determined by three different structural loci, as indicated by the genetic variation of the electrophoretic hemoglobin pattern observed in natural populations. Hemoglobins occur that, despite identical chain compositions, have different electrophoretic mobilities. This may be due to a mechanism, known from man, where part of the hemoglobin is blocked by a hexose.


Genetic polymorphism in the bladder campion, Silene maritima. A survey of protein variability has been made in wild populations of Silene maritima, a perennial outcrossing coastal plant. In the most extensively studied populations, at Beesands, six out of 21 loci were polymorphic, and the average heterozygosity was 0.153. Four polymorphic loci were studied on an extensive shingle beach at Dungeness. Three loci showed no frequency cline with distance from the sea; a fourth locus, that for adenylate kinase, showed a barely significant cline, which was not confirmed by a repeat study in the same place a year later. Five polymorphic loci were studied at Beesands along a strip of shingle beach and up a contiguous cliff. No frequency difference between the two habitats was observed. There are significant differences in allelic frequencies between localities. It is concluded that the data fail to show an association between habitat and gene frequency, although they were collected in such a way as to be capable of demonstrating such an association if it existed.


Hb Mobile [alpha2beta2 73(E17)Asp replaced by Val]: a new variant. A new hemoglobin variant found in a mother and her child was characterized by column chromatography of the tryptic hydrolysate of the aminoethylated, glycinamidated beta-chain, followed by chymotryptic digestion of the abnormal beta T-9 peptide and amino acid analyses. It was shown to be alpha2beta2 73(E17)Asp replaced by Val and named by Hb Mobile.


Simple vs. complex inheritance of inducible aryl hydrocarbon hydroxylase in mouse tissues. The genetics of induction of hepatic and lung aryl hydrocarbon hydroxylase (AHH) have been studied in Af/Ki and AKR/Ki mice and in their F1 and F2 progeny after administration of 3-methylcholanthrene (3MC). Furthermore, the induction of AHH was investigated using the fetal liver explant model system with 3MC, trans-1,2-dihydroxy-3MC, and 4'-bromoflavone as the inducers. The results obtained with the above strains were contrasted with those from the C57BL/6Ki, DBA/2+Ki, and their crosses. The present investigation revealed a complex pattern of inheritance of basal and inducible AHH in lung and liver of AKR/Ki and Af/Ki, with a poor correlation between lung and liver. Hepatic AHH was not fully inducible in the F1 hybrids, while the frequency distribution function in the F2 mice was suggestive of more than two distinct classes.


Gene activation of alcohol dehydrogenase in Japanese quail and chicken-quail hybrid embryos. No preferential activation of the maternally derived alcohol dehydrogenase (ADH) allele was found in any of the chicken male x Japanese quail female hybrids examined. ADH activity in the liver was, in fact, found to exist in two different cathodal zonal regions on starch gel electropherograms; the zone II bands appeared at day 5 of incubation in the quail embryo (day 6 in the hybrid embryo) and the zone I bands appeared in 9-day quail embryos (10-day hybrid embryos). By day 13 of incubation, only the faster-migrating zone I bands could be detected in both quail and hybrid embryos.


Citrulline metabolism in normal and citrullinemic human lymphocyte lines. Citrullinemia is one of the five aminoacidurias associated with the Krebs-Henseleit urea cycle. A long-term lymphocyte line (UM-21) derived from a patient with this disease and nine of ten clones of this line were found to have no activity for the enzyme argininosuccinate synthetase (AS), as demonstrated by their inability to grow in medium in which citrulline had been substituted for arginine, by their inability to incorporate arginine-C14 derived from citrulline-C14 into cellular protein, and by direct enzyme assay. One clone had normal or nearly normal argininosuccinate synthetase activity, as demonstrated by the same criteria. Nutritional "variants" able to grow logarithmically in medium containing citrulline were isolated from UM-21 and three clones. The apparent Kms of AS for citrulline in UM-21, the ten clones, the variant lines, and a normal line were measured and fell into three groups: AS in UM-21 and nine clones had no measurable apparent Km for citrulline; AS in the variant cells had apparent Kms for citrulline of approximately 20 mM; and AS in the normal cell line and one clone had apparent Kms for citrulline of 0.2 mM. The data suggest that the defect in the citrullinemic cell lines is due to a mutation in the structural gene coding for argininosuccinate synthetase.


Allozyme genetics in permanent translocation heterozygotes of the Oenothera biennis complex. Allozyme inheritance and transmission genetics of 11 enzyme systems were determined in the permanent translocation heterozygotes Oenothera biennis, Oe. strigosa, and Oe. parviflora. Electrophoretic variation was examined first among 164 strains of structural heterozygotes. Allelic configurations were then judged from inheritance patterns in reciprocal F1 hybrids between each of 22 ring-forming strains and tester strains of the related bivalent-formers, Oe. hookeri and Oe. grandiflora. Allozymes are inherited as codominant markers, and, as dictated by the genetic system, within a strain individual allelic variants are generally transmitted through only one germ line. Of the 20 loci resolved, only eight are polymorphic in any species, and, within species, generally only two alleles are present at each polymorphic locus. Despite the relatively meager allelic array, each of the 22 strains whose chromosome complexes were characterized is genotypically unique. Generally, within taxa, alpha (egg) and beta (sperm) complexes differ in allele frequency at several polymorphic loci. Such variability is correlated with differences in the phylogenetic origins of complexes and not with differences in segmental arrangement within a group of related complexes.


Induction of 16alpha-hydroxylase in cultured human lymphocytes. Lymphocytes grown for 72 hr in the presence of mitogenic agents exhibited a nine-fold increase in 16alpha-hydroxylase activity over a 24-hr period following the addition of 17beta-estradiol as an inducing agent. The major metabolite of 17beta-estradiol produced by induced lymphocytes showed a chromatographic mobility indistinguishable from that of estriol. Enzymatic activity increased linearly with cell numbers, was inhibited by CO, and exhibited no induced increase in the presence of cycloheximide. A population survey indicated about 68% of a randomly selected Caucasian group to be essentially uninducible for 16alpha-hydroxylase.


Ribonucleic acid synthesis in isolated rat liver nuclei under conditions of ribonucleic acid processing and transport. A cell-free system is described which permits a significant and prolonged synthesis of RNA in isolated rat liver nuclei, under conditions previously demonstrated to support normal nuclear processing and transport of both rRNA and mRNA. The system contains cytosol but not (NH4)2SO4 or other non-physiological components. Evidence is presented for cytosol factors which stimulate ribosomal, and to a lesser degree, non-ribosomal RNA synthesis.


Interaction of bilirubin with reconstituted collagen fibrils. 1. Interaction of bilirubin with collagen fibrils was explored in a two-phase system where collagen was present as an opaque rigid gel composed of striated fibrils, and bilirubin as an aqueous solution. 2. The Ka value of the binding of bilirubin to collagen fibrils is 5.4 X 10(3)M-1. The interaction of bilirubin with collagen fibrils depends on temperature. Below 5 degrees C, the binding is greatly diminished and denaturation of collagen fibril aggregates at 52--53 degrees C into a dissolution state abolishes binding of bilirubin. 3. Salicylate and sulphanilamide do not affect the binding of bilirubin to reconstituted collagen fibrils. 4. Serum albumin (40--80mM), known to reverse the binding of bilirubin to lipids, dissociates only 50% of the bilirubin bound to collagen fibrils. This suggests that sites located on collagen participate in some tight binding of bilirubin and the corresponding binding sites on albumin do not compete with them. 5. Urea (4M) abolishes more than 70% of the binding of bilirubin to collagen. Urea and thermal denaturation studies indicate the importance of conformation and organization of collagen fibrillar aggregates for the binding of bilirubin.


Analysis of the forms of acetylcholinesterase from adult mouse brain. The solubilization of 80% of the acetylcholinesterase activity of mouse brain was performed by repeated 2h incubations of homogenates at 37 degrees C in an aqueous medium. Analysis of the soluble extract by gel filtration on Sephadex G-200 showed that up to 80% of the enzyme activity was eluted in a peak which was estimated to consist of molecules of about 74000mol.wt. This peak was called the monomer form of the enzyme. After 3 days at 4 degrees C, the soluble extract was re-analysed and was eluted from the column in four peaks of about 74000, 155000, 360000 and 720000 mol.wt. Since the total activity of the enzyme in these peaks was the same as that in the predominantly monomer elution profile of fresh enzyme, we concluded that the monomer had aggregated, possibly into dimers, tetramers and octomers. Extracts of the enzyme were analysed by polyacrylamide-gel electrophoresis and the resulting multiple bands of enzyme activity on gels were shown to separate according to their molecular sizes, that is by molecular sieving. All these forms had similar susceptibilities to the inhibitors eserine, tetra-isopropyl pyrophosphoramide and compound BW 284c51 [1,5-bis-(4-allyldimethylammoniumphenyl)pentan-3-one dibromide]. Thus the forms of the enzyme in mouse brain which can be detected by gel filtration and polyacrylamide-gel electrophoresis may all be related to a single low-molecular-weight form which aggregates during storage. This supports similar suggestions made for the enzyme in other locations.


Separation of a series of chromophores and fluorophores present in elastin. Purified elastin was hydrolysed with HCl and manipulated under conditions that minimized oxidation. Gel-permeation chromatography on polyacrylamide gel and ion-exchange chromatography on dextran cation-exchanger each resulted in the separation of a series of yellow fluorescent fractions. These hitherto unreported ampholytes have fluorescence spectra that approximate to that of the intact protein, and account for its characteristic optical properties. Since the coloured fluorophores are confined to enzyme-resistant regions of the protein molecule they appear to have important structural implications.


The actin content of fibroblasts.

The actin content of fibroblasts. Cultures of chick skin fibroblasts were dissolved in solutions of sodium dodecyl sulphate, and their entire protein content was examined by gel electrophoresis. The most abundant species migrated in the same position as muscle actin. It gave a similar pattern of iodinated peptides after reaction with radioactive sodium iodide and digestion with proteinases, and contained comparable amounts of Nt-methylhistidine. Its amount was estimated by quantitative densitometry of stained gels with bovine serum albumin as an internal standard, and by radioactive assay of cultures that had been grown in the presence of [35S]methionine. The values obtained ranged from 7 to 14% of the total cellular protein, with an average of 8.5%. A protein band in the position of muscle myosin was also present and accounted for about 2.5% of the total protein. Both this and the actin band increased in relative amount with the age of the cultures.


The interaction between bovine serum albumin and surfactants. 1. Potassium n-decyl phosphate binds exothermically to bovine serum albumin at pH 7.0 to form a specific complex containing approx. 60 phosphate anions. 2. The formation of the complex is accompanied by changes in the u.v. difference spectrum of the protein. 3. At higher phosphate concentrations (above 0.4mM) surfactant molecules continue to be bound, and the protein undergoes a gross change in conformation. 4. n-Dodecyltri-methylammonium bromide binds endothermically to bovine serum albumin at pH7.0 but the extent of binding for a given free surfactant concentration is less than for the phosphate surfactant. 5. Binding is accompanied by a small change in the specific viscosity and by changes in the u.v. difference spectrum of the protein. 6. It is suggested that over the surfactant concentration ranges studied n-decyl phosphate ions first bind to the C-terminal part of the protein and then to the more compact N-terminal part whereas n-dodecyltrimethylammonium ions bind only to the C-terminal part of bovine serum albumin.


Decarboxylation of uroporphyrin by heating at atmospheric pressure. Free uroporphyrin is decarboxylated by heating in the air to 150--200 degrees C. Complete decarboxylation occurs after heating to 200 degrees C for 5 min, with a yield of 66--75%. Heptacarboxylic porphyrin can be prepared with a yield of 40% by heating uroporphyrin to 150 degrees C for 60 min, and hexacarboxylic porphyrin with a yield of about 20% by heating to 150 degrees C for 2h.


Some properties of the citrate synthase from the extreme halophile, Halobacterium cutirubrum. 1. Citrate synthase [citrate oxaloacetate-lyase (CoA-acetylating), EC 4.1.3.7] was purified about 400-fold from the extreme halophile, Halobacterium cutirubrum, by a method involving (NH4)2SO4 fractionation, chromatography on DEAE-cellulose and hydroxyapatite and gel filtration on Sephadex G-200. 2. The purified enzyme was best activated by high concentrations of KCl (3M); the chlorides of other cations and K+ salts of other anions (Br-, NO3-, SCN-) were less effective than KCl as activators. The enzyme was best stabilized by high concentrations of NaCl or KCl. Cold-lability was found in the presence of 3M-KCl, but not in the presence of NaCl at concentrations up to 5M. The results suggest that both the shielding of negative charges on the enzyme molecule and the stabilization of hydrophobic bonds by high KCl concentrations were required for maximum activity of the enzyme. 3. The double-reciprocal plots for acetyl-CoA or oxaloacetate at several concentrations of the co-substrate intersected at the abscissa in the presence of either KCl or NaCl, at either 1 or 3M. The Km for oxaloacetate increased about fivefold with the salt concentration, from 1 to 3M.


The sequences of the coenzyme-binding peptide in the cytoplasmic and the mitochondrial aspartate aminotransferases from sheep liver. The sequences of the coenzyme-binding peptide of both cytoplasmic and mitochondrial aspartate aminotransferases from sheep liver were determined. The holoenzymes were treated with NaBH4 and digested with chymotrypsin; peptides containing bound pyridoxal phosphate were then isolated. One phosphopyridoxyl peptide was obtained from sheep liver cytoplasmic aspartate aminotransferase. Its sequence was Ser-Ne-(phosphopyridoxyl)-Lys-Asn-Phe. This sequence is identical with that reported for the homologous peptide from pig heart cytoplasmic aspartate aminotransferase. Two phosphopyridoxyl peptides with different RF values were isolated from the sheep liver mitochondrial isoenzyme. They had the same N-terminal amino acid and similar amino acid composition. The mitochondrial phosphopyridoxyl peptide of highest yield and purity had the sequence Ala-Ne-(phosphopyridoxyl)-Lys-Asx-Met-Gly-Leu-Tyr. The sequence of the first four amino acids is identical with that already reported for the phosphopyridoxyl tetrapeptide from the pig heart mitochondrial isoenzyme. The heptapeptide found for the sheep liver mitochondrial isoenzyme closely resembles the corresponding sequence taken from the primary structure of the pig heart cytoplasmic aspartate aminotransferase.


The amino acid sequence of plastocyanin from spinach. (Spinacia oleracea L.). The amino acid sequence of spinach (Spinacia oleracea L.) plastocyanin was determined. It consists of a single polypeptide chain of 99 residues and has a sequence molecular weight of 10415. The sequence was determined by using a Beckman 890C automatic sequencer and by the dansyl--phenyl isothiocyanate analysis of peptides obtained by the enzymic digestion of purified CNBr fragments. Overlap through the two methionine residues was not shown. Sedimentation equilibrium in the ultracentrifuge gave a molecular weight for spinach plastocyanin of about 9000, in contrast with the value of 21000 reported previously by Katoh et al. (1962).


Purification of porcine enterokinase by affinity chromatography. A method is described for the purification of porcine enterokinase by affinity chromatography with p-aminobenzamidine as the ligand. Purification was completed by immunoadsorption with antisera raised to components binding non-biologically to the gel. The final enterokinase preparation was 2.3 times more active than the most active preparation previously described.


The use of ammonium acetate in the precipitation of ribonucleic acid. Ammonium acetate (0.24M) in combination with cold 66% (v/v) ethanol quantitatively precipitated RNA from very dilute solutions (greater than or equal to mug/ml) after centrifugation for 2.5 X 10(6)g-min. There was also less co-precipitation of detergents with ammonium acetate than NaCl.


The distinguishing characteristics of the plasma membrane are its exposed proteins. The exposed proteins of the plasma membrane of normal human lymphocytes and platelets were labelled by using the lactoperoxidase macromolecular probe system. The labelled components were separated into molecular-weight classes by sodium dodecyl sulphate--polyacrylamide-gel electrophoresis. In contrast with the report by Tanner et al. (1974), a comparison of the two cell types showed that the major labelled components in both cell types were glycoproteins and were not identical. It is concluded that the exposed proteins are probably the most distinguishing characteristic of the plasma membrane of differentiated cell types.


Nature of the multiple forms of sweet-potato glucose phosphate isomerase. The previously reported isoenzymes of sweet-potato glucose 6-phosphate isomerase were resolved by DEAE-cellulose chromatography. The multiple forms exhibited identical electrophoretic properties and electrofocused as a single component with an apparent isoelectric pH of 4.0. Chromatographic studies also suggest that the multiple forms do not represent true isoenzymes.


The use of the direct linear plot for determining initial velocities. A new method is described for estimating initial velocities of enzyme-catalysed reactions. It is simple to apply either graphically or numerically, and is particularly appropriate for experiments in which the initial straight part of the progress curve is very short or non-existent. It requires no more knowledge than is readily available about the details of the system, such as the extent of reaction at equilibrium, the rate of enzyme inactivation, the nature of product inhibition etc., unlike some other methods of analysing progress curves, which are often invalidated by small errors in the defining assumptions.


Sigmoid curves, non-linear double-reciprocal plots and allosterism. 1. The theory of plane curves was applied to the graphical methods used in enzyme kinetics and a mathematical analysis of the possible graph shapes is given. 2. The belief that allosterism can be inferred from steady-state data alone is subjected to criticism and the mathematical significance of sigmoid curves and non-linear double-reciprocal plots is explored. 3. It is suggested that the usual methods of interpreting steady-state kinetic data are often based on over-restrictive assumptions which prevent maximum utilization of the available data. 4. Methods for obtaining the degree of the rate equation from graph shapes obtained directly from initial-rate measurements and from replots of asymptotic behaviour as chi approach the level 0 and chi approach the level infinity are discussed. 5. Detailed proofs of the theorems given in the text have been deposited as Supplementary Publication SUP 50049 (10 pages) at the British Library (Lending Division), Boston Spa, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1975), 145, 5.


Properties of protein-chlorophyll complexes from pea (Pisum sativum L.) leaves. The organization of chlorophyll. Chlorophyll-protein-detergent complexes were prepared from pea chloroplasts by using sodium dodecylbenzenesulphonate and polyacrylamide-gel electrophoresis. Circular-dichroism spectra showed that complex CPI has a dimeric arrangement of chlorophyll a, with additional weaker interactions. Ellipticities were determined for both complexes and for purified chlorophylls in solution, and it is argued that the circular dichroism of complex CPII is derived from chlorophyll-protein interaction rather than from interaction between chlorophylls a and b. The detergent could be removed from the complexes by using urea and gel filtration, leaving the chlorophyll-protein in solution, although in each case a diminished ellipticity indicated some loss of organization. Three-peaked circular-dichroism spectra of chloroplast fragments before and after addition of detergent were compared with a curve obtained by summing graphically the spectra of complexes CPI, CPII and the free-pigment fraction. There was good correspondence at 650 nm, and the longer-wavelength peaks agreed in form and magnitude, but with discrepancies in position. It was concluded that complexes CPI and CPII pre-exist in the original material, but that there is an environmental effect which is destroyed when the complexes are extracted.


The active site of penicillinase from Staphylococcus aureus PC1. Isolation of a specific covalent complex with the substrate quinacillin. 1. The penicillinase-catalysed hydrolysis of quinacillin was quenched by addition of 5 m-guanidinium chloride or 1% (w/v) sodium dodecyl sulphate, and the quenched reaction mixture was dialysed exhaustively against solutions of the denaturant. 2. Irreversibly bound quinacillin was shown by titration with HgCl2 to be covalently attached to the protein by the beta-lactam carboxyl group. 3. The derivative was found to be stable over the pH range 3.5-8.5. 4. Chymotryptic hydrolysis of the product and subsequent fractionation showed that quinacillin was bound to one or possibly two peptides.


Stopped-flow fluorescence studies on saccharide binding to lysozyme. The binding of the beta-1-4-linked trimer of N-acetyl-D-glucosamine to hen egg-white lysozyme was studied by rapid-reaction-kinetic methods with tryptophyl fluorescence observation of the transients. It was found that discrete segments of the fluorescence-difference spectrum from this reaction were perturbed at different time-points during the binding process. The results were interpretated as the formation of the initial complex, the fast phase of the reaction, perturbing the environment of tryptophan-62 and a subsequent and slower rearrangement of the initial complex perturbing the environment of tryptophan-108. At pH 4.4, the release of protons from aspartate-101 occurred during the rearrangement step of the binding reaction. A model for the reaction is presented (E, enzyme; L, ligand): (see article) The association of this ligand with lysozyme may be visualized in three-dimensional terms as initial complex-formation across the top of the active-site cleft followed by a diving motion of the ligand into the cleft.


Physicochemical properties of Planorbis corneus erythrocruorin. The erythrocruorin from the snail Planorbis corneus had a sedimentation coefficient, so/20,w, of 33.5 +/- 0.31 S, and a molecular weight of 1.65 x 10(6) +/- 0.04 x 10(6) by high-speed sedimentation-equilibrium ultracentrifugation. The amino acid composition and absorption spectrum of the protein are reported. A very low number of half-cystine residues was found, corresponding to 0.4 residue per haem group. The haem content was 2.76 +/- 0.22%, corresponding to a protein molecular weight of about 22300. Under both acid and alkaline conditions partial dissociation took place to yield mixtures of products that could not be identified. A subunit corresponding to that containing one haem group was not obtained under any of the dossociating conditions tried. Electron microscopy revealed a ring-shaped molecule about 12.2 +/- 0.5 nm in diameter. The native erythrocruorin bound O2 co-operatively, the intermediate value of h in Hill plots having values between 1.7 and 3.4 depending on the conditions.


Studies of the acetylcholinesterase from houseflies (Musca domestica L.) resistant and susceptible to organophosphorus insecticides. Acetylcholinesterase from the heads of insecticide-resistant and -susceptible houseflies (Musca domestica L.) was studied in vitro. The enzymes could not be distinguished electrophoretically, and their behaviour on polyacrylamide-disc-gel electrophoresis was influenced by the presence of Triton X-100 in both the homogenate and the gels. In the absence of detergent, the acetylcholinesterase was heterogeneous, but behaved as a single enzyme when it was present. By analogy with studies of acetylcholinesterase from other sources, these observations were attributed to aggregation of the enzyme when not bound by membranes. The enzyme from resistant flies was more slowly inhibited than the susceptible enzyme, bimolecular rate constants (ki) differing by approx. 4-20-fold for a range of organophosphorus compounds. The kinetics of inhibition of acetylcholinesterase were consistent with the results of electrophoresis, i.e. they corresponded to those of a single enzyme in the presence of Triton X-100, but a mixture of enzymes in its absence. The susceptibility of the more sensitive components in these mixtures was determined.


X-ray photoelectron spectra of iron-sulphur proteins. The X-ray photoelectron spectra of the 2p, 3s and 3p levels of iron in oxidized Clostridium pasteurianum ferredoxin indicate that the eight iron atoms in the molecule are indistinguishable. Their magnetic state is indicated both by core polarization splitting of the 3s electrons, and by "shake-up' satellites on the 2p lines. Similar satellites are observed in the 2p lines of reduced Chromatium high-potential iron-sulphur proteins and oxidized spinach ferredoxin, indicating that there too the iron atoms are magnetic. The low observed magnetic susceptibility of these proteins is therefore due to spin-coupling between the iron atoms in the active centre.


Secondary alkylsulphatases in a strain of Comamonas terrigena. The occurrence in a strain of Comamonas terrigena of secondary alkylsulphatase activity towards potassium decan-5-yl sulphate is reported. A number of cell-washing and osmotic-shock procedures for releasing bacterial exocytoplasmic enzymes were ineffective in releasing this activity. Primary alkylsulphatases are not present in the organism, nor can their formation be induced under a wide variety of experimental conditions tested.


Acetyl groups in cell-wall preparations from higher plants. O-Acetyl groups were detected by i.r. spectroscopy in cell-wall preparations from grasses and other higher plants and their presence was confirmed chemically. The amounts present are likely to influence both the physical state of the cell-wall polysaccharides and also their digestion by enzymes.


The identification of a peptide in human parotid saliva particularly active in enhancing the glycolytic activity of the salivary micro-organisms. A factor in saliva responsible for markedly activating the glycolytic activity of micro-organisms was isolated from parotid secretions and identified as a small basic peptide. Analysis of the peptide showed a high proportion of histidine, lysine and arginine. Its minimum molecular weight was calculated to be between 2500 and 3000.


The amino acid sequence of troponin I from rabbit skeletal muscle. The complete amino acid sequence of rabbit skeletal muscle troponin I was determined by the isolation of the cyanogen bromide fragments and the tryptic methionine-containing peptides. Troponin I contains 179 amino acid residues and has a molecular weight of 20864. Its N-terminus is acetylated. Detailed evidence on which the sequence is based has been deposited as Supplementary Publication SUP 50055 (23 pages) at the British Library (Lending Division), Boston Spa, Wetherby, West Yorkshire LS23 7QB, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1975) 145, 5.


The Sulphation of p-hydroxyphenylpyruvic acid and related compounds by the rat liver cytosol. Cytosol preparations of rat liver and kidney were examined for their ability to transfer sulphate from adenosine 3'-phosphate 5'-sulphatophosphate to p-hydroxyphenylpyruvic acid. Little activity towards this substrate was observed, and the main product detected in the reaction mixtures was identified as p-hydroxybenzyl alcohol O-sulphate. This was not formed from p-hydroxybenzaldehyde, a spontaneous oxidation product of p-hydroxyphenylpyruvic acid, by sulphation followed by a rapid enzyme-catalysed reduction of the intermediate phydroxybenzaldehyde O-sulphate. This product was formed mainly by this sequence of reactions, but the reverse, reduction followed by sulphation, also appeared possible. p-Hydroxybenzyl alcohol itself was very readily sulphated by both preparations, and the liver also produced a sulpho-conjugate of homogentisic acid. These observations are important in calculating the turnover of L-tyrosine O-sulphate in the mammalian system, and establish that p-hydroxyphenylpyruvic acid O-sulphate is an end product of its metabolism, rather than an intermediate in its synthesis by reversed transamination.


Interactions of glucose, acetoacetate and insulin in mammary-gland slices of lactating rats. 1. Utilization of 5mM-glucose by slices of lactating mammary gland was decreased 33% on addition of acetoacetate (2mM) to the incubation medium. This inhibition was accompanied by increases in the intracellular concentrations of citrate and glucose 6-phosphate. 2. In the presence of acetoacetates the accumulation of pyruvate in the medium approximately doubled. 3. Insulin completely reversed the inhibitory effect of acetoacetates on glucose utilization, without altering the amount of acetoacetate removed or pyruvate formed. 4. Similar results were obtained with mammary-gland slices from diabetic rats, except that insulin did not completely reverse the effects of acetoacetates. 5. Acetoacetate inhibited the formation of 14CO2 from [1-14C]pyruvate; this effect was not overcome by insulin. 6. Insulin increased the proportion of [3-14C]acetoacetate that was converted into lipid and decreased that oxidized to CO2.7. The physiological significance of these findings is discussed.


Rate-limiting factors in the oxidation of ethanol by isolated rat liver cells. The oxidation of ethanol by isolated liver cells from starved rats is limited by the rate of removal of reducing equivalents generated in the cytosol by alcohol dehydrogenase. Evidence is presented suggesting that, in these cells, transfer of reducing equivalents from the cytosol to the mitochondria is regulated by the intracellular concentrations of the intermediates of the malate-aspartate and glycerol 3-phosphate cycles, as well as by flux through the respiratory chain. In liver cells isolated from fed rats, the availability of substrate increased the cell content of intermediates of the hydrogen-transfer cycles, and enhanced ethanol uptake. Under these conditions, ethanol consumption is limited by the availability of ADP for oxidative phosphorylation.


Betaine-induced stimulation of respiration at high osmolarities in a halotolerant bacterium. It was shown that non-penetrating solutes at high concentrations inhibit the respiration of the halotolerant bacterium Ba1. Betaine relieved the inhibition caused by osmotic stress and exhibited in this respect a considerable structural specificity. The rate of oxidation of various substrates was stimulated to different extents. It stimulated the rates of both respiration and growth to a similar extent, leaving the energy yield essentially unchanged. In cells pre-loaded with labelled glutamate, betaine also stimulated the rate of oxidation of this intracellular substrate. Betaine was accumulated by respiring cells, and the maximum amount taken up was correlated with the osmolarity of the medium. As judged by chromatography, accumulated intracellular betaine underwent no chemical modification, and this accumulated betaine could not be exchanged with the betaine in the medium or released by passive efflux when respiration was inhibited. Intracellular betaine caused no stimulation of respiration, whereas betaine added to the medium increased the respiratory rate to the same extent in cells pre-loaded with betaine as that in the nonloaded cells. The above observations suggest that iso-economic adjustment is not involved in the anti-osmotic effect of betaine, and that betaine exerts its action on the cellular membrane from the outside.


Skeletal-muscle growth and protein turnover. Because of turnover, protein synthesis and breakdown can each be involved in the regulation of the growth of tissue protein. To investigate the regulation of skeletal-muscle-protein growth we measured rates of protein synthesis and breakdown in growing rats during development on a good diet, during development on a marginally low-protein diet and during rehabilitation on a good diet after a period of severe protein deficiency. Rates of protein synthesis were measured in vivo with a constant intravenous infusion of [14C]tyrosine. The growth rate of muscle protein was measured and the rate of breakdown calculated as breakdown rate=synthesis rate-growth rate. These measurements showed that during development on a good diet there was a fall with age in the rate of protein synthesis resulting from a fall in capacity (RNA concentration) and activity (synthesis rate per unit of RNA). There was a fall with age in the breakdown rate so that the rate was highest in the weaning rats, with a half-life of 3 days. There was a direct correlation between the fractional growth and breakdown rates. During rehabilitation on the good diet, rapid growth was also accompanied by high rates of protein breakdown. During growth on the inadequate diet protein synthesis rates were lesss than in controls, but growth occurred because of decreased rates of protein breakdown. This compression was not complete, however, since ultimate muscle size was only one-half that of controls. It is suggested that increased rates of protein breakdown are a necessary accompaniment to muscle growth and may result from the way in which myofibrils proliferate.


Drug induction and sex differences of renal glutathione S-transferases in the rat. Treatment of male rats with 3,4-benzopyrene, 3-methylcholanthrene and phenobarbital resulted in the induction of glutathione S-aryl- and S-aralkyl-transferase activities in kidney cytosol. Benzopyrene produced 77 and 44% increases in aryl and aralkyl activities respectively. Methylcholanthrene caused 73 and 86% increases in the retrospective activities, whereas phenobarbital treatment increased only aralkyl activity (51%). There was no effect on epoxide or alkyl glutathione S-transferase activities with these treatments. Differences were found between the specific activities of the four glutathione S-transferases in females and males, with the following female/male ratios: aryl 0.74; aralkyl 2.37; epoxide 1.52; alkyl 1.33. No changes in Km values were observed relative to drug induction or sex differences. Comparisons are made between the findings of this report and corresponding experiements with liver.


Biochemical changes in cultured foetal rat liver explants. Liver explants from 20-day-old foetuses cultured for 48h in the absence of serum released 70% of their total soluble protein content into the medium. In the presence of serum this loss still amounted to 60%. The concentration of total particulate protein remained unchanged but there was some translocation of mitochondrial enzymes to the cytosol, and enzymes expected to increase during this stage of development failed to do so. The addition of cortisol plus glucagon (to serum-containing media) did not decrease the loss of total soluble protein from the explants but induced considerable tyrosine aminotransferase activity which was not released into the medium. The observations suggest that under the usual culture conditions a minority of the cells retain their functional integrity. The extent of deterioration, not reflected in histologically visible necrosis or cell damage, can be conveniently monitored by the malate dehydrogenase activity released to the medium.


Characteristics of aspartate deamination by the puring nucleotide cycle in the cytosol fraction of rat liver. 1. The component reactions of the puring nucleotide cycle were studied in cytosol extracts of rat liver. 2. AMP deaminase was strongly activated by ATP and analogues of ATP. 3. IMP was converted into ATP by a system requiring the presence of aspartate, GTP and a nucleoside triphosphate-regenerating system. 4. Under appropriate conditions, NH3 was produced from aspartate. 5. From the rates at which these reactions occur it is concluded that the purine nucleotide cycle may have sufficient activity to be a major pathway of amino acid deamination in liver.


The effect of inhibitors in vivo on protein synthesis and the amino acid pool in the sheep blowfly, Lucilia cuprina. 1. The rates of detoxification of cycloheximide (33 mug/g fresh wt.), puromycin (167 mug/g fresh wt.) and actinomycin D (1 mug/g fresh wt.) were assessed in vivo on the basis of acid-insoluble [14C]leucine incorporation in the sheep blowfly, Lucilla cuprina; these were compared with quantitative estimates which took account not only of incorporation data but also of leucine pool size and turnover. Quantitatively, cycloheximide and puromycin were still at least 50% effective in inhibiting protein synthesis after 6.5 and 24.5h of exposure respectively, whereas values based only on incorporation data suggested that cycloheximide was 83% effective and puromycin completely ineffective after the respective periods. Quantitative estimates also showed that actinomycin D effectiveness increased with increasing exposure over 24.5h, in contrast with values based only on incorporation data, which suggested that it was completely ineffective after 24h.2. All inhibitors affected the dynamic state of the amino acid pool; there was a marked decrease in the rate of leucine-pool turnover as well as an increase in the half-life of leucine in the pool. 3. Inhibition of protein synthesis resulted in changes in leucine-pool size; the most pronounced increase occurred with cycloheximide and puromycin and the most pronounced decreases with actinomycin D. 4. Evidence is presented which suggests that proteolysis is functionally linked to protein synthesis, which determines its rate indirectly.


Conversion of [U-14C]threonine into 14C-labelled amino acids in the brain of thiamin-deficient rats. In confirmation of the findings of Gaitonde et al. (1974), a decrease in the brain concentration of threonine and serine, and an increase in glycine, were observed in rats maintained on a thiamin-deficient diet. Similar changes were found in the blood, and the concentration of several other amino acids in the blood decreased significantly. There was a correlation between the concentrations of threonine, serine, aspartate and asparagine in the brain and blood. In experiments in which [U-14C]threonine was injected into rats most of the radioactivity in the brain and blood of control rats was, as expected, in threonine in the acid soluble metabolites. In contrast, a considerable proportion of radioactivity was also found in other amino acids, namely glutamate, glutamine, aspartate, gamma-aminobutyrate and alanine, in the brain of thiamin-deficient rats. [U-14C]Threonine was also converted into 14C-labelled lactate and glucose, but the extent of this conversion was severalfold higher in thiamin-deficient than in control rats. This finding gave evidence of the stimulation in thiamin-deficient rats of the catabolism of [U-14C]threonine to [14C]lactate by the aminoacetone pathway catalysed by threonine dehydrogenase, and into succinate via propionate by the alpha-oxobutyrate pathway catalysed by threonine dehydratase (deaminase). The measurement of specific radioactivities of glutamate, aspartate and glutamine after injection of [U-14C]threonine, indicated a stimulation of the activities of threonine dehydrogenase and threonine dehydratase (deaminase) in the brain of thiamin-deficient rats. The specific radioactivities of glutamate, asparatate and glutamine int he brain were consistent with an alteration in the metabolism of threonine, mainly in the 'large' compartment of the brain of thiamin-deficient rats. The measurement of relative specific radioactivity of proteins after injection of [U-14C]threonine indicated a marked decrease in the synthesis of proteins, mainly in the liver of thiamin-deficient rats.


Glutamate uptake by chick retina.

Glutamate uptake by chick retina. The uptake of glutamate was found to be via a single high-affinity transport mechanism with Km values of 35 and 95 mum for chick-embryo and mature chick retina respectively. These data contrast with the uptake of gamma-aminobutyratewhich in the same tissue has previously been shown to display two kinetically distinct mechanisms in the embryo, but a single low-affinity process in the mature retina.


Characteristics of normal labour in different racial groups. The characteristics of normal labour in 1306 white, Asian and black parturients have been established following a prospective study of 3217 consecutive labours. Asian patients were found to be of significantly shorter stature than white or black women (p less than 0-001) and their infants significantly lighter than those of white (p less than 0-001) and of black (p less than 0-05) women, and a low positive correlation was found between maternal height and infant birth weight. The mean duration of the first stage of labour, taken from the time of admission to the labour ward, was 5-6 hours in primiparae and 3-7 hours in multiparae. The mean durations of the second stage of labour were 41-5 and 17-4 minutes respectively. The correlations between the duration of the first and second stages of labour were too low to be of value in patient management. Similar low correlations were found between the duration of the second stage of labour and both infant birth weight and the Apgar score at one minute. Cervical dilatation-time curves, constructed with reference to the cervical dilatation found on admission to the labour ward, revealed no significant differences in the progress of normal labour in the different racial groups.


Plasma sex steroid and gonadotrophin patterns in human menstrual cycles. Sensitive and specific displacement analysis methods for the assay of steroid hormones in small volumes of plasma are described. Plasma sex steroid and gonadotrophin hormone patterns were determined throughout a number of normal menstrual cycles. The mean cycles showed patterns which were similar to those described by other workers. However, examination of individual cycles provided information which may contribute to our understanding of menstrual cycle regulation with particular reference to the pattern of 17alpha-hydroxyprogesterone and steroid-gonadotrophin interactions.


Infertility in women with apparently ovulatory cycles. I. Comparison of their plasma sex steroid and gonadotrophin profiles with those in the normal cycle. Serial daily plasma profiles of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone, 17alpha-hydroxyprogesterone and oestradiol 17beta throughout single menstrual cycles from seven patients with a history of infertility but with normal menstrual rhythm and no apparent ovarian dysfunction were compared with those from apparently normally menstruating women. This comparison showed that the infertile group produced poor corpora lutea after poor follicular growth. The poor follicular growth may be related to depressed pituitary release of FSH in response to a negative feedback effect of elevated progesterone levels.


Infertility in women with apparently ovulatory cycles. II. The effects of clomiphene treatment on the profiles of gonadotrophin and sex steroid hormones in peripheral plasma. Six infertile patients had been studied previously (Dodson et al., 1975b) and were shown to produce inefficient corpora lutea which appeared to be predetermined by ovulation of "poorly grown" follicles. In a second cycle these infertile patients were treated with 50 mg of clomiphene per day for five days. The resulting plasma sex steroid and gonadotrophin profiles were compared with those found before treatment and with the profiles in normal patients (Dodson et al., 1975a). Treatment with clomiphene appeared to produce increased follicular growth and more active corpora lutea.


The analysis of continuous fetal heart rate traces in the first and second stages of labour. In the search for a simplified method of fetal heart rate trace assessment 50 traces recorded during the first stage of labour and those recorded in the last hour of each of a further 50 labours have been analysed intensively. The importance of baseline variability in predicting possible fetal distress in both the first stage and the last hour of labour is shown and the possible use of an objective measurement of baseline variability in the further screening for fetal acidosis is described. The importance of lag time measurement in the first stage of labour is demonstrated and the relationships between fetal scalp blood pH and other heart rate parameters is discussed. Significant inverse correlations between one-minute Apgar score and maximum amplitude of deceleration and total deceleration area in the last hour of labour are also shown. Temporal relationships between intrauterine pressure rises and transient changes of fetal heart rate failed to give significant correlations in the last hour. These findings in the last hour of labour are compared with those obtained in the first stage of labour and the differences are discussed.


Changes in the renin-angiotensin system in primigravidae with hypertensive disease of pregnancy. Plasma renin concentration, renin activity and angiotensin II (AII) have been studied in 35 primigravidae at or near term immediately prior to surgical induction of labour. A highly significant positive correlation was established between diastolic blood pressure at the time of sample collection and plasma AII. This relationship was observed over the range of diastolic pressure studied irrespective of the diagnosis made at the time of admission to hospital. In patients with hypertension, a significant negative relationship was established between plasma renin concentration and renin activity and AII. Furthermore, plasma AII levels were higher for a given plasma renin activity or plasma renin concentration in the hypertensive women than in the normotensive group. These findings suggest that the renin-angiotensin system is actively involved in hypertensive disease of pregnancy and that suppressed levels of renin concentration and activity may result from elevated angiotensin levels. It is suggested that the utero-placental complex may be the source of the elevated AII levels.


The benefits and risks of amniocentesis for the assessment of fetal lung maturity. Amniocentesis for estimation of the lecithin: sphingomyelin (L/S) ratio was performed on 483 patients on 552 occasions. The introduction of this test has been associated with a highly significant (p less than 0-001) decrease of 59 per cent in the incidence of respiratory distress syndrome. The finding of a low L/S ratio (less than 2-0) enabled delivery to be deferred until lung maturity had been reached in 43 (10-3 per cent) of the 419 patients who had successful amniocentesis. Fetal death occurred after amniocentesis in two patients (0-41 per cent). One fetus died after haemorrhage occurred behind an anterior placenta and the other died of exsanguination. Failed amniocentesis occurred in 64 patients (13-2 per cent), and was more common when the placenta was anterior or was not located before the procedure and when fetal growth was retarded. Fetal distress in labour occurred more commonly after failed amniocentesis. Spontaneous labour followed amniocentesis in 47 patients (9-7 per cent) and was significantly more prone to occur after 38 weeks gestation. Rhesus isoimmunization did not occur in any of the rhesus-negative patients who delivered rhesus-positive babies and who were screened six or more months after amniocentesis, although anti-D immunoglobulin was not given routinely after amniocentesis. Suggestions are made as to how the success rate of this valuable procedure can be improved and how some of the serious fetal and maternal sequelae can be avoided.


Uterine action after induction of labour with oral prostaglandin E2 tablets compared with intravenous oxytocin. Uterine action throughout the whole of labour induced by low amniotomy and either oral prostaglandin E2 tablets or intravenous ocytocin was studied in 10 randomly matched pairs of patients. Oral prostaglandin E2 tablets induced fewer contractions per hour, and these contractions were longer and more variable in length than those induced by intravenous oxytocin. There was no difference in basal uterine tone or in the amplitude of contractions. A comparison was made between the outcome of labour induced by low amniotomy and either oral prostaglandin E2 tablets 0-5 mg. hourly in 65 patients, or intravenous ocytocin in 41 patients. This dose of prostaglandin E2 tablets was an inadequate adjunct to low amniotomy in nulliparous patients. Despite this, the induction-delivery interval for the whole prostaglandin group was not significantly greater than that for the oxytocin group. There were no significant differences in fetal heart rate pattern, or in the incidence of gastro-intestinal side-effects between the two groups.


Recurrent bleeding from the lower segment scar--a late complication of Caesarean section. Hysterectomy became necessary six months after Caesarean section because of recurrent severe haemorrhage from the uterine scar.


An improved large scale fractionation of high mobility group non-histone chromatin proteins. 1. Methodology is presented for the large scale preparation and fractionation of high mobility group proteins from calf thymus chromatin. The total high mobility group protein from approx. 1 kg calf thymus tissue can be separated into five fractions by CM-Sephadex C25 ion-exchange chromatography. High mobility group proteins 1 and 2 comprise two fo the fractions. From a third fraction two more chromatin proteins, protein 3 and 17, can be isolated by trichloroacetic acid precipitation and CM-cellulose chromatography at pH 5.5. 2. The four proteins thus purified are lysine-rich proteins. Proteins 1 and 2 are additionally characterised by their high contents of acidic amino acids, as described previously (Goodwin, G. H. and Johns, E. W. (1973) Eur. J. Biochem. 40, 215-219). Proteins 3 and 17, having lower contents of acidic amino acids, are basic proteins similar to the histones. All four proteins exhibit single N-terminal amino acids; glycine is the N-terminal group of proteins 1, 2 and 3; protein 17 has a proline N-terminal amino acid. The proteins are not highly phosphorylated nor are they associated with appreciable quantities of nucleic acid.


Oblique banding pattern in collagen fibrils reconstituted in vitro after trypsin treatment. Collagen fibres from rat tail tendon suspended in small pieces in a solution (pH 7.8) containing 0.5 M CaCl2 were treated with purified bovine trypsin at 20 degrees C for 20 h. After the enzyme treatment collagen from this solution was precipitated out and reconstituted in vitro into native-type fibrils. The banding pattern in these reconstituted fibrils was found to be oblique. This is comparable to that observed recently in fibrils reconstituted from cartilage collagen. On the other hand, normal transverse banding pattern was observed in the fibrils reconstituted in vitro from collagen solution of rat tail tendon which was not pre-treated with trypsin. No significant change was, however, observed in the segment long spacing fibrils precipitated from the enzyme-treated collagen solution. It is possible that the enzyme might affect the mode of organization of tropocollagen molecules during in vitro fibrillogenesis into native-type fibrils either by interacting with the "telopeptide" regions or with the non-collagenous components associated with the native protein and this could probably result into the formation of fibrils with oblique banding pattern.


On the active streaming experiment of actomyosin solutions in glass microcapillaries. The active-streaming experiments of Oplatka et al. (Oplatka, A. and Tirosh, R. (1973) Biochim. Biophys. Acta 305, 684-688 and Oplatka, A. Gadasi, H., Tirosh, R. Lamed, Y., Muhlrad, A. and Liron, N. (1974) J. Mechanochem. Cell Motil. 2, 295-306) with actomyosin solution in a glass microcapillary is reexamined under various conditions with several kinds of reference material. It is found that vigorous streaming took place in the actomyosin solution as reported by Oplatka et al. However, streaming which is indistinguishable from that observed in the actomyosin solution in the presence of actomyosin ATPase activity also occurred, even when the ATPase activity was blocked. The streaming also cannot be confirmed as being active when using acto-heavy meromyosin solution. There is a possibility that the streaming experiment provides interesting information on the microscopic state of solutions which is not directly related to the chemo-mechanical conversion.


9 S thyroid particulate iodoprotein.

9 S thyroid particulate iodoprotein. Particulate iodoproteins have been studied in the rat thyroid gland using the isotopic 125I equilibration method. Pulse experiments were also performed with a second isotope, 131I. Labelled iodoproteins, both soluble and solubilized by digitonin from the thyroid particulate material, were analyzed by sucrose-gradient ultracentrifugation. 1. At isotopic equilibrium and irrespective of the iodide content of the diet two particulate iodoproteins, with sedimentation coefficients of 27 and 19 S, were solubilized by digitonin. In addition a 9 S iodoprotein was also present but its proportion varied markedly with the iodine content of the diet: it accounted for 50-60% of the label found at the particulate level when the dialy diet of iodine was high (75-500 mug/day) but was almost absent when the diet was only 2 mug/day. 2. Most of this 9 S protein was found in the 600-15 000 times g particulate pellet, i.e. a fraction enriched in lysosomes and phagolysosomes. 3. The iodoamino acid composition of the 9 S fraction was very similar to that of the 19 S particulate thyroglobulin: its hormone content was 19%. 4. The double precipitation technique showed that the 9 S fraction is immunochemically related to thyroglobulin. 5. Pulse experiments showed that the 9 S particulate iodoprotein was slowly labelled by 131I. 6. The amount of 9 S iodoprotein was increased by thyrotropin (30-40% increase versus control experiments 5 min after thyrotropin injection). These properties of the 9 S particulate iodoprotein are discussed in relation to the assumption that it might be a product of partial proteolysis of thyroglobulin after endocytosis and partial digestion by the phagolysosomes.


Evidence for the formation of an ester between thrombin and heparin cofactor. Heparin cofactor, a thrombin inhibitor, is purified from human plasma by affinity chromatography on heparin-agarose. The nature of the binding between thrombin and the inhibitor is studied by treatment of the complex with 6 M guanidinium chloride, hydroxylamine, and dilute alkali. The complex is not dissociated during gel chromatography in 6 M guanidinium chloride. This result supports an earlier proposal that formation of the complex includes the formation of a covalend bond. Treatment of dodecylsulfate-denatured complex with hydroxylamine results in dissociation of the complex to yield free thrombin and heparin cofactor. The complex is also dissociated in dilute NaOH (pH 12) solutions. These results indicate that the covalent bond between thrombin and the inhibitor is a carboxylic ester.


Properties of a hemoglobin from the chromatrope of the nematode Mermis nigrescens. The chromatrope pigment of Mermis nigrescens (Phylum: Aschelminthes, Class: Nematoda) was previously thought to have a role in photoreception. In this work it is shown to be an oxyhemoglobin whose absorption spectrum in vivo and in extracts resembles that of Ascaris oxyhemoglobins in having a weak alpha-band absorptivity. The extracted hemoglobin binds O2 and CO reversibly and with an affinity higher than that of the same concentration of horse hemoglobin. The Soret band of the deoxy derivative is unusually low and broad. Absorptivities and lambdamax for absorption bands of the oxy, deoxy and CO derivatives are tabulated for comparison with other hemoglobins. Microchemical procedures were developed which revealed that the chromatrope contains an average of 5-10(-12) mol of non-dialysable protoheme. Thus the hemoglobin concentration in the approx. 0.5 nl chromatrope volume is on the order of 10 mM (heme). The O2 binding ability and high in vivo concentration of this hemoglobin make possible a role in O2 storage or facilitation of O2 diffusion.


Characterization of the collagen of human hypertrophic and normal scars. The collagen produced in response to an injury of human skin is initially stabilized by a cross-link derived from hydroxyallysine, and characteristic of embryonic skin. In normal healing there is a change over with time to the cross-link derived from allysine, which is typical of young skin collagen. In contrast, hypertrophic scars fail to follow the time-related changes of normal skin, but retain the characteristics of embryonic collagen, indicating a continued rapid turnover of the collagen. This is further supported by the high proportion of the embryonic Type III collagen present in hypertrophic scars.


Studies on in vitro polymerization of tubulin from renal medullary extracts. Previous in vivo studies showed that microtubules are involved in the cellular action of vasopressin. In order to analyze the role of renal medullary microtubules, a system was developed which would allow the study of the assembly of tubulin in renal medulla extracts into microtubules in vitro. The assembly of tubulin into microtubules occurred in renal medullary cytosol (100 000 times g supernatant) under specific conditions which include pre-concentration of cytosol by ultrafiltration, the presence of ethylene glycol bis(2-aminoethyl)ether tetraacetic acid (EGTA) and 4 M glycerol, and warming at 37 degrees C. Formation of microtubules, which sedimented at 100 000 times g, was proved by (a) an increase in the apparent [3H]colchicine-binding activity of depolymerized pellets, (b) appearance of typical microtubules as shown by electron microscopy, and (c) by the increase in the quantity of microtubular protein analyzed by polyacrylamide gel electrophoresis. Vinblastine at a concentrationof 10(-6) M completely blocked formation of microtubules. A slight increase of ionized calcium in the polymerization mixture also prevented microtubule assembly; this inhibitory effect of ionized calcium was present at concentrations as low as 10(-4) M. Blockade of microtubule assembly by the increase in concentration of ionized calcium or by vinblastine may be the basis of known inhibitory effect of these two agents upon the hydroosmotic effect of vasopressin in vivo.


Non-histone chromosomal proteins easily extractible from chick erythrocytes. Those non-histone chromosomal proteins which are easily extractible from chick erythrocytes differ substantially from proteins similarly extracted from other tissues of various species. Although a protein P1 was isolated along with histone H1 by extraction of calf thymus chromatin with HC1O4, the same procedure did not extract this protein from chick erythrocyte chromatin of either normal or regenerating blood. Likewise , non-histone proteins extracted with 0.35 M NaCl from calf thymus differed from those of normal chick erythrocytes, which were qualitatively identical but quantitatively inferior to those of regenerating blood. The major protein of about 25 000 molecular weight, totally extracted with 0.35 M NaCl from calf thymus, was not found in chick erythrocyte chromatin, but rather another major protein of about 35 000 molecular weight was partially extracted from erythrocytes.


Comparison of the predicted and observed secondary structure of T4 phage lysozyme. Predictions of the secondary structure of T4 phage lysozyme, made by a number of investigators on the basis of the amino acid sequence, are compared with the structure of the protein determined experimentally by X-ray crystallography. Within the amino terminal half of the molecule the locations of helices predicted by a number of methods agree moderately well with the observed structure, however within the carboxyl half of the molecule the overall agreement is poor. For eleven different helix predictions, the coefficients giving the correlation between prediction and observation range from 0.14 to 0.42. The accuracy of the predictions for both beta-sheet regions and for turns are generally lower than for the helices, and in a number of instances the agreement between prediction and observation is no better than would be expected for a random selection of residues. The structural predictions for T4 phage lysozyme are much less successful than was the case for adenylate kinase (Schulz et al. (1974) Nature 250, 140-142). No one method of prediction is clearly superior to all others, and although empirical predictions based on larger numbers of known protein structure tend to be more accurate than those based on a limited sample, the improvement in accuracy is not dramatic, suggesting that the accuracy of current empirical predictive methods will not be substantially increased simply by the inclusion of more data from additional protein structure determinations.


Enzymic and immunochemical properties of lysozyme. XI. Conformation and immunochemistry of the two-disulfide peptide and the tryptophan and lysine residues in its antigenic reactivity. The previously described peptide 62-68 (Cys 64-Cys 80) 74-96 (Cys 76-Cys 94) (Atassi, M.Z., Suliman, A.M. and Habeeb, A.F.S.A. (1975) Biochim. Biophys. Acta 405, 452-463), which accounted for about one-third of the total antigenic reactivity of native lysozyme, was isolated here with lysine 97 attached to it. The peptide was subjected to specific modification reactions in order to determine some of the residues which formed part of its antigenic reactive site. ORD measurements showed that the peptide was greatly unfolded in solution relative to its expected mode of folding within the intact lysozyme molecule. Modification of the two tryptophan residues in the peptide by reaction with 2,3-dioxo-5-indolinesulfonic acid provided a derivative which possessed similar conformational parameters to those of the unmodified peptide. However, the derivative retained only about half the immunochemical reactivity of the peptide. Succinylation of the amino groups afforded a derivative whose conformational parameters were identical to those of the unmodified peptide but in which half of the immunochemical reactivity was lost. Modification of the two tryptophan residues followed by succinylation of the amino groups resulted in almost complete loss of the antigenic reactivity, and the loss was not due to conformational differences. The antigenic reactivity of the peptide was also destroyed on removal of tryptophans 62 and 63, of sequence 84-93 from the loop 74-79 and of sequence 74-75 by chymotryptic digestion. From these and previous results it was concluded that the antigenic reactive site in this part of the lysozyme molecule incorporates one or both of tryptophans 62 and 63 as well as one or both lysines 96 and 97. The two disulfides 64-80 and 76-94 bring these two parts of the lysozyme molecule into a single reactive site. The intactness of the disulfides is essential for maintenance and reactivity of the site.


Covalent structure of fibrinopeptides from buffaloes breeding in Italy. The primary structure of fibrinopeptides A and B from buffaloes breeding in Italy has been determined with a view to establishing whether this animal is an autochthonous species or is the result of recent radiation mutation. Some differences exist at the morphologic and physiologic levels between the Indian buffalo and that breeding in Italy, But they do not allow a clear evolutionary line to be traced between these species. The amino acid sequences, if compared with homologous sequences of the Indian buffalo, show one difference in the most variable region of fibrino-peptides A, in particular the substitution of a serine residue by a glycine residue in position 8. This difference supports the hypothesis of the autochthonous origin of the Italian buffalo.


On the nature of the cytosine-methylated sequence in DNA of Bacillus brevis var. G.-B. On growing the cells of Bacillus brevis S methionine-auxotroph mutant in the presence of [Me-3H]methionine, practically all the radioactivity incorporated into DNA is found to exist in 5-methylcytosine and N6-methyladenine. The analysis of pyrimidine isopliths isolated from DNA shows that radioactivity only exists in mono- and dinucleotides and the content of 5-methylcytosine in R-m5 C-R and R-m5 C-T-R oligonucleotides is equal. The analysis of dinucleotides isolated from DNA by means of pancreatic DNAase hydrolysis allows the nature of purine residues neighbouring 5-methylcytosine to be identified and shows that 5-methylcytosine localizes in G-m5 C-A and G-m5 C-Tr fragments. B. brevis S DNA methylase modifying cytosine residues recognizes the GCA/TGC degenerate nucleotide sequence which is a part of the following complementary structure with a two-fold rotational axis of symmetry: (5')...N'-G-C-T-G-C-N... (3') (3')...N-C-G-A-C-G-N'... (5') (Methylated cytosine residues are askerisked). Cytosine-modifying DNA methylase activity is isolated from B. brevis cells; it is capable of methylating in vitro homologous and heterologous DNA. Hence DNA in bacterial cells can be undermethylated. This enzyme methylates cytosine residues in native and denatured DNA in the same nucleotide sequences. Specificity of methylation of cytosine residues in vitro and in vivo does not depend on the nature of substrate DNA. DNA methylases of different variants of B. brevis (R, S, P+, P-)) methylate cytosine residues in the same nucleotide sequences. It means that specificity or methylation of DNA cytosine residues in the cells of different variants of B. brevis is the same.


Effect of chloramphenicol on early mRNA synthesis in bacteriophage lambda. The N gene product induces lambda-delayed early mRNA synthesis. This transcription was sensitive to chloramphenicol at the very early stage of phage development, but became resistant within 2-4 min after infection. However, the same transcription was sensitive to the antibiotic throughout phage development if lambda tof- mutant was used as the infecting phage. The results suggest involvement of the tof gene in lambda early transcription; the tof product may act as a stabilizer of the N gene function.


The macrophage and host defense mechanisms. Macrophages participate in a variety of host defense mechanisms. The delivery of macrophage precursors from the bone marrow to the tissues may be reduced with bone marrow failure and macrophage function may be impaired with certain virus infections such as herpes zoster or infectious mononucleosis. Defective macrophage function may impair cell mediated immunity more than humoral immunity which is manifested clinically by cutaneous anergy. One disease associated with chronic macrophage dysfunction is chronic mucocutaneous candidiasis.


Role of macrophages in immunity, with special reference to tumour immunology. A review. It seems that macrophages, which are present in most tissues, are involved in many steps of the immune reaction. They are essential for initiating the response and they assist the lymphocytes in several ways. By themselves, they play an important role in the immune surveillance and in tumour immunology. The behaviour of macrophages is strongly influenced by lymphocytes. However macrophages are equipped with a special machinery, including an active membrane and a developed lysosomal system. A lot remains to be done on its functions and on the trigger mechanisms.


Ventricular compliance and ageing.

Ventricular compliance and ageing. Compliance of the right ventricle has been studied in 29 aged subjects who were free from any overt heart failure. The observations indicate that a loss of ventricular compliance results, in aged subjects, in an impairment of diastolic filling, which itself is responsible for a reduction in cardiac output. However, there appears to be no relationship between ventricular compliance and systolic performance but the failure to find any such relation may be due to a contractile adaptation in subjects having a low ventricular compliance.


[Influence of hypothyroidism on the lacticodehydrogenase isoenzymes during the cerebral cortex maturation in the rat]. The isoenzymes of lacticdehydrogenase of cerebral cortex, investigated by starch gel electrophoresis, are changed during pre and neonatal thyroidectomies in relation to these factors: a) dose of the drug (131I or Propylthiouracil), b) date of gestation, c) the age of the animal (with a maximal modification at the 17th-18th day). They retain an enzymatic pattern like the fetal pattern, showing the retardation of the developing central nervous system.


Dangers and problems in calculating coefficients of a sum of exponential functions. The use of the computer in making calculations has led increasingly to the estimation of parameters of exponential functions from point experimental measurements. This involves the use of techniques such as logarithmic transformations and criteria (especially that of the least squares), of which the use is not always justified. Radioactivity measurements raise special problems as they are subject to a statistical distribution of the Poisson type. The authors propose a method based on the statistical criterion of "maximum likelihood" which permits tests of the number of exponentials from point measurements (in practice, this method is valid for one or two exponentials), and also the calculation of parameters more satisfactorily than customary methods.


A computer-based record and organisation system for a department of urology. This paper describes the development and practical application of a computer-based record system in a urological department. It demonstrates how such a system can provide a clinical flow sheet for day-to-day use combined with a computerised data store for both clinical evaluation and for research purposes. The paper also outlines how the system has improved patient handling and general departmental efficiency.


Acute hypertension and perirenal haematoma--a new aspect of Page's syndrome. Chronic hypertension induced by perirenal constrictive processes is known from experimental studies as well as from clinical experience. In this report we present 2 cases of acute hypertension induced by massive perirenal haematoma, in 1 case as a complication following pyeloplasty, in the other as a complication following percutaneous renal biopsy. Since the shock may be masked by the hypertension (and operation thereby postponed) it is essential to be aware of this syndrome.


The angiographic evaluation of renal pelvic carcinoma. The angiographic findings are described in 6 cases of renal pelvic carcinoma. An angiographic finding which has not hitherto been emphasised is a diminished branching of arteries in the kidney segment related to the pelvi-calyceal tumour--the "pruned-tree" appearance. This is generally associated with a deficiency of the nephrogram in the same segment. These appearances may be due to invasion of the renal parenchyma by tumour. However, the findings may occur also in the absence of parenchymal invasion by the tumour and therefore do not necessarily reflect the stage of the disease.


Some observations and theories on the wide ureter and hydronephrosis. A simple but effective classification of the wide ureter is proposed. It separates ureters into two major groups of refluxing and non-refluxing, but emphasises that not all non-refluxing ureters are obstructed. An analysis is made of normal and abnormal ureteric peristalsis and the part it plays in conducting a bolus through the pelviureteric and ureterovesical junctions. A theory is advanced to explain the poor emptying of the wide ureter and the abnormal pelviureteric junction. It is based on the inability of the circular and longitudinal muscles to form a bolus and it explains how the unopposed and inadvertent contraction of the circular muscle grips the orifice in each situation to aggravate the poor emptying.


Procidentia and ureteric obstruction.

Procidentia and ureteric obstruction. 3 cases of renal failure, due to procidentia, presenting as emergencies are described. The upper urinary tracts of 4 asymptomatic patients with procidentia were investigated; dilatation was found in 1 instance only.


The value of intestinal valves in urinary diversion--an experimental study. It has been shown in experimental animals that an ileorectal valve incorporated into the operation of uretero-ileosigmoidostomy cannot be relied upon to prevent ascending urinary tract infection. However, a colorectal valve, not previously described, has been effective in preventing the development of a metabolic acidosis in piglets in whom this method of urinary diversion has been carried out.


Malignant phaeochromocytoma of the urinary bladder--report of two cases. 2 new patients with malignant phaeochromocytoma of the urinary bladder were presented. 4 other cases with the same disease reported in literature were reviewed and discussed.


False cytodiagnosis of bladder malignancy due to previous radiotherapy. Results from a urological cytodiagnosis service indicate that it is not possible to distinguish with any degree of certainty malignant from non-malignant irradiated urothelial cells in urine. It is stressed, therefore, that when urine specimens are sent for cytological examination, details of any previous irradiation should be given to the cytologist.


Carcinoma of the bladder complicating cyclophosphamide therapy. Two patients with cyclophosphamide induced cystitis and co-existent bladder tumour are described. Possible mechanisms for these associated diseases are discussed and the difficulty of diagnosing bladder tumour in patients with haemorrhagic cystitis is stressed.


Non-parasitic chyluria (presenting with urinary retention). Two cases of non-parasitic chyluria are presented each presenting with retention of urine. Abnormal communications in both cases have been demonstrated lymphangiographically. The thoracic ducts were patent and there was no evidence of filariasis. Surgical removal of the anomalous dilated lymphatics was carried out, the patients being symptom-free at 7 and 10 years. The two cases seem to have lymphatic malformations which may be the basis of the disorder.


The urodynamic assessment of adult enuresis. Urodynamic analysis has revealed a high proportion of uninhibited bladder activity in adult enuresis. There was no associated neurological deficit to explain this abnormality. There was no correlation between the various diurnal symptoms and the presence of uninhibited bladder contractions. Enuresis was associated with obstructive uropathy in only 6 percent of cases. Some evidence has also been presented to support the concepts of ectopic sensory receptors and abnormal sphincter inhibition. A review of the problem of pathogenesis has suggested four main areas of possible abnormality: 1. A sensory lesion producing inadequate or delayed appreciation of bladder activity. 2. Cortical and subcortical dysfunction because of inadequate arousal or defective maturation of subconscious inhibition. 3. A congenital or acquired upper motor neurone lesion causing an uninhibited bladder. 4. A similar or related lesion causing an over-inhibited sphincter. These factors probably always occur in varying combinations. Because of the lack of association between symptoms, signs and objective measurements, there is a strong case for relating diagnosis, prognosis and treatment to urodynamic assessment.


Hydrocolpos.

Hydrocolpos. The method of presentation and problems in diagnosis of 3 cases of hydrocolpos are discussed. The anatomy and embryology of the abnormality is described. The methods of treatment and results are outlined.


Evolution of the felid brain.

Evolution of the felid brain. Endocranial casts of 15 genera of fossil felids provide a record of felid brain evolution over the past 35 million years. Brains of the earliest felids, known as paleofelids, had coronolateral, suprasylvian and variably developed ectosylvian sulci as their only neocortical sulci. The last paleofelids, which became extinct around 8 million years ago, show little change in external brain morphology except for the addition of a presylvian sulcus. The other group of felids, the neofelids, appears about 25 million years ago, with coronolateral and suprasylvian sulci their main neocortical sulci, plus a discontinuous ectosylvian sulcus and small postlateral, sylvian and presylvian sulci. The posterior cerebellar vermis was straight and unexpanded. Beginnings of expansion of the sigmoid gyri and development of the cruciate sulcus are evident in neofelids 15-20 million years ago, and by 5-9 million years ago neofelids had brains that appear modern in external morphology. Endocasts of four genera of Pleistocene saber-toothed felids are similar in sulcal pattern to those of modern felids, except for Dinobastis, which had a unique expansion of visual cortex. Endocasts of 27 species of modern felids, representing the six commonly recognized genera, are strikingly similar in external morphology, although the brains of a few species, such as cheetahs, lynxes and jagouarundis, display distinguishing features. Modern felid brains average about the same size relative to body weight as do those of viverrids, but are about 25% smaller in relative size than those of canids. Olfactory bulbs are relatively smaller in felids than in canids or viverrids.


Effect of phenols on the oxidation of estradiol by uterine peroxidase. The effect of various phenols on the conversion of [4 -14C]estradiol to water-soluble products by estrogen-induced uterine peroxidase (EC 1.11.1.7) has been investigated. Evidence was provided that those phenols which enhanced the oxidation of estradiol exerted their effect by activating peroxidase or protecting the enzyme from inactivation by the products of the reaction rather than by inhibiting the breakdown of hydrogen peroxide by catalase (EC 1.11.1.6). It has also been shown that tyrosine acted both as an activator of uterine peroxidase and as a water-soluble acceptor for the metabolites of estradiol. The ability of tyrosyl peptides to form conjugates with estradiol was influenced by the other amino acids and decreased with the number of adjacent tyrosyl residues.


Cross-linking of hemoglobin, haptoglobin, and hemoglobin-haptoglobin complex with bifunctional imidoesters. Dimethyl adipimidate was used to cross-link the polypeptides within hemoglobin, haptoglobin, and hemoglobin-haptoglobin complex. Cross-linked hemoglobin retained considerable ability to bind haptoglobin, although the amounts bound were reduced and the haptoglobin reaction could be used to fractionate the modified hemoglobin. With cross-links limited to intramolecular sites, hemoglobin showed four bands on polyacrylamide gel electrophoresis in sodium dodecyl sulfate, identified, with reference to the subunit polypeptides, as monomer, dimer, trimer, and tetramer. The dimer region consisted of at least two separable species. When hemoglobin-haptoglobin complex was cross-linked, a band of hemoglobin dimer was present, which demonstrates that at least two hemoglobin subunits have a close spatial relation when bound to haptoglobin. Some comparisons with adipimidate-reacted hemoglobin were made using malonimidate and suberimidate and some marked differences were noted.


In vitro metabolism of 17beta-estradiol by human liver tissue. 17beta-[6,7- 3H]Estradiol was incubated with adult human liver slices in Krebs-Ringer phosphate buffer containing glucose. Of the identified 3H recovered, 51-76 percent consisted of estrone-3-sulfate (E13S) and 17 beta-estradiol-3-sulfate (E23S). E13S was the main metabolite and was found in both tissue and medium. E23S was present only in the medium. Minor amounts of estrogen glucuronides were formed. When a human liver homogenate was incubated with [3H]E2 in a medium fortified with excess uridine diphosphate glucuronic acid only some 4 percent of conjugation with glucuronic acid was observed. It is suggested that human liver favors sulfurylation as the conjugating mechanism for E2 and E1.


The tris-catalyzed isomerization of potassium D-glucose 6-O-sulfate. Potassium D-glucose 6-O-[35S]sulfate was partially converted by Tris (greater than or equal 25 mM, pH 7.5) to 35SO4(2-), fructose, and a 35S-labelled compound which was tentatively identified as fructose 6-O-sulfate. The isomerization reaction was also catalyzed by ethanolamine and 2-dimethylaminoethanol but not by glycine, triethanolamine, or pentaerythritol. The significance of these findings in relation to kinetic studies of the enzyme glycosulfatase is discussed.


Essential sulfhydryl groups of rat liver monoamine oxidase. The inhibition by some thiol reagents of partly purified mitochondrial monoamine oxidase (MAO) (EC 1.4.3.4) from rat liver was studied, and the molar content of sulfhydryl groups in the enzyme determined. Sodium nitroprusside and 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) inhibited the enzyme, apparently reversibly, while sodium arsenite was not inhibitory. Concentrations of the respective inhibitors causing 50% inhibition after 15 min of preincubation with the enzyme at pH 7.0 and 37 degrees C are 5.80 times 10(-4) M and 4.35 times 10(-5) M. The thiol compounds cysteine, dithiothreitol, and 2-mercaptoethanol did not inhibit MAO. The average number of sulfhydryl groups per mole of enzyme, determined by reaction with DTNB, increased from 3.6 +/- 0.2 freely reacting sulfhydryl groups (n = 4) to 18.4 to total sulfhydryl groups (n = 2) on denaturation with 8 M urea.


Antibody status to poliomyelitis, measles, rubella, diphtheria and tetanus, Ontario, 1969-70: deficiencies discovered and remedies required. A serologic survey was made in 15 health unit areas, testing some 5000 individuals in the age groups 4 to 6, 11 to 13, 15 to 17 and 23 to 45 years. Two types of serious deficiency were found. Only 65% of children 4 to 6 years old had antibodies to all three types of poliovirus, the antibodies being due almost entirely to immunization with Salk vaccine. Even in children who had had six or more doses only 74% had antibodies to the three types. The high percentage of students 11 to 13 and 15 to 17 years old with poliovirus antibodies can be attributed largely to natural infection and to Sabin vaccine in the mass campaign of 1962, as well as to Salk vaccine. In children who had received Sabin vaccine as well as Salk vaccine a very high level of immunity was found. The immunity of the school-age population will decline to an insufficient level unless Sabin vaccine is used after immunization with Salk vaccine. Of children 4 to 6 years old 18% had no diphtheria antitoxin and 6% had no tetanus antitoxin. Even in those who had had six or more doses of the antigens 5% had no diphtheria antitoxin and 1 to 2% had no tetanus antitoxin. This apparently refractory state is probably due to the use of unadsorbed toxoids, and it is clear that adsorbed toxoids should be used. In the adults, diphtheria antitoxin was found in only 55% and tetanus antitoxin in only 38%.


Neonatal hypertyrosinemia and evidence for deficiency of ascorbic acid in Arctic and subarctic peoples. Hypertyrosinemia tyrosine concentration in whole blood greater than 0.42 mmol/l or 7.5 mg/dl is prevalent among lnuit newborn of the Canadian Eastern Arctic. The rate was 14.8 per 100 newborn between January 1970 and December 1972 (first survey period) and 6.2/100 between January 1973 and September 1974 (second survey period); the corresponding rates among Indian newborn of Nouveau Quebec were 2.6 and 2.2%. Among Anglo-Saxons the rate was less than 0.5% and in French Canada it commonly exceeded 0.94%. Serum concentrations of ascorbic acid were low (less than or equal to 0.25 mg/dl) in the pregnant and age-matched adult lnuit when measured by Nutrition Canada during the first survey period. The percentages of inuit children (up to 4 years old) and pregnant women at "high risk" for scurvy (serum concentration of ascorbic acid less than 0.2 mg/dl) were 14.8 and 47.1, respectively; the corresponding national percentages were 3.0 and 2.2, respectively. Deficiency of ascorbic acid in pregnant women is probably the cause of the unusual prevalence of neonatal hypertyrosinemia among the native Arctic and subarctic peoples because ascorbic acid is required to maintain optimal activity of p-hydroxyphenylpyruvic acid hydroxylase and to permit normal oxidation of tyrosine.


Prevalence of recurrent herpes labialis and aphthous ulcers among young adults on six continents. The prevalence of recurrent herpes labialis (RHL) and recurrent aphthous ulcers (RAU) in young adults - - 635 armed-forces recruits and 9897 health-profession students - - in 48 institutions in 21 countries was determined by a questionnaire survey. Two or more occurrences (lifetime prevalence) of RHL were reported by 33.2% of men and 28.0% of women; the corresponding figures for RAU were 38.7% and 49.7%. North American respondents, mainly from Canada, had a significantly higher prevalence of both lesions. There were some differences in relation to profession. Approximately 15% of all the people surveyed had had herpes labialis and 25% had had aphthous ulcers at least once during the previous year. Persons with a history of recurrence of one lesion were more likely to have a history of recurrence of the other.


What is a medical experiment?

What is a medical experiment? Innovative therapy may appear to coincide with medical experimentation, raising ethical and legal issues, for instance on informed consent and institutional review. Medical treatments may be classified, however, to distinquish novel procedures from experimentation.


Atheromatous embolic disease.

Atheromatous embolic disease. In five patients with atheromatous embolic disease, the diagnosis was made before death in four -- on the basis of cholesterol emboli in the retina in three and from renal pathologic features in 1. Muscle biopsy demonstrated emboli in one patient, and emboli were seen in the vessels of amputated toes in two. All patients died of renal failure, but there was evidence of multisystem involvement in addition. Autopsy in four cases showed characteristic cholesterol emboli in many organs.


[Anaplastic epithelioma of the thyroid gland 6 years after cervical irradiation for malignant lymphoma]. Carcinoma of the thyroid gland may develop after irradiation of the cervical region. It is well documented in patients who received radiation therapy to the neck in childhood but rarely when the therapy was given in adulthood. A case of an undifferentiated carcinoma of the thyroid gland occurring in an adult 6 years after irradiation of the cervical region for a malignant lymphoma is reported. The role of irradiation and the factors related to the development of carcinoma of the thyroid gland are reviewed.


Porphyria cutanea tarda: comparison of cases precipitated by alcohol and estrogens. A group of seven patients with porphyria cutanea tarda (PCT) precipitated by excessive alcohol consumption (A) was compared with a group of nine patients with PCT precipitated by estrogen therapy (B). Comparison was based on clinical signs, biochemical and morphologic evidence of liver disease, results of serum iron studies and response to therapy. Group A patients were men of mean age 57 years; group B patients were women of mean age 39 years who had been taking estrogen orally, either for contraception (in combination with progesterone) or as replacement therapy. Clinical signs were essentially the same in the two groups. Some patients in both groups had biochemical and morphologic evidence of liver disease. Group A patients had elevated values for serum iron and total iron-binding capacity, whereas patients in group B had normal or low values. Cessation of estrogen therapy of less than a year's duration brought about a spontaneous clinical and biochemical remission in group B patients. Otherwise, phlebotomy seemed to be the therapy of choice in both groups.