The removal of amateur tattoos by salabrasion.

Due to the recent upsurge in self-inflicted tattoos in teenagers it was decided to employ for their removal the method of salabrasion, i.e. abrasion with table salt. This technique was recently used by Manchester (1973, 1974) for the removal of commercial tattoos. Fourteen patients with twenty-eight tattoo or only tiny flecks of pigment remaining visible; eleven showed a fair response, i.e. the tatoo, although much lighter in colour, was still legible but the patient was satisfied with the result; three showed a poor response, i.e. the tattoo appeared as if untreated; four (two patients) defaulted. In general, as could be expected, the darker the tattoo the more difficult was its removal and those on fingers were particularly resistant.


Specific hyposensitization in atopic dermatitis.

Specific hyposensitization is rarely advocated for the treatment of atopic dermatitis. Good results have been obtained in fifteen mainly adolescent and adult patients selected from over 3,000 patients with atopic dermatitis. Criteria included a history of exacerbations of the dermatitis after exposure to the antigen, an airborne antigen which could not be avoided, a rather distinctive clinical picture and symptoms bad enough to warrant the considerable difficulties involved.


Trachyonychia (rough nails).

The term trachyonychia is a useful one to describe roughening of the surface of many nails due to various causes. Psoriasis and alopecia areata are responsible for some cases. Lichen planus can roughen nails but the change is usually limited to a few nails and accompanied by discomfort. In many cases no cause can be found and these cases are somewhat arbitrarily listed either as twenty nail dystrophy of childhood or severe nail dystrophy.


Yellow nails and nephrotic syndrome.

A patient is described who simultaneously developed the yellow nail syndrome and nephrotic syndrome. Treatment of the nephrotic oedema coincided with return of normal nail growth.


Complement abnormalities in diffuse plane xanthomatosis with paraproteinaemia.

Paraproteinaemia may be associated with xanthomatous skin deposits and these can arise in the absence of elevated lipid levels. Two cases of benign monoclonal gammopathy with diffuse plane xanthomatosis are reported. Case 1 exhibited hypolipidaemia and a functional deficiency of C1 esterase inhibitor. Case 2 showed a normal lipoprotein profile, abnormal platelet aggregation, and a cutaneous vasculitis with evidence of complement consumption via the classical pathway. The significance of these abnormalities is discussed.


The involvement of the lipid phase transition in the plasma-induced dissolution of multilamellar phosphatidylcholine vesicles.

Unsonicated liposomes prepared from dimyristoyl phosphatidylcholine were nearly completely dissolved during a 3 h incubation with rat plasma at or close to the phase-transition temperature of 24 degrees C. At 37 or 15 degrees C virtually no liposomal disintegration was observed even after 24 h of incubation. The liposomal solubilization, which was monitored by turbidity measurements or by determination of phospholipid sedimentability, was accompanied by the formation of a phospholipid-protein complex similar or identical to the one we previously reported to be formed from sonicated liposomes of egg phosphatidylcholine (Scherphof, G., Roerdink, F., Waite, M. and Parks, J. (1978) Biochim. Biophys. Acta 542, 296--307). Unsonicated multilamellar liposomes made of egg phosphatidylcholine were completely resistant to the dissolving potency of plasma when incubated at 37 degrees C. Liposomes from equimolar mixtures of dimyristoyl and dipalmitoyl phosphatidylcholine were only degraded by plasma in the temperature range between 30 and 35 degrees C at which temperature this cocrystallizing phospholipid mixture undergoes a phase transition. However, even at these temperatures the rate of dissolution of this mixture was significantly lower than of dimyristoyl phosphatidylcholine at 24 degrees C. In the dissolving process of this mixture a slight preference for the lower-melting component was observed. The ability of cholesterol to completely abolish the susceptibility of dimyristoyl phosphatidylcholine liposomes to plasma at a 1:2 molar ratio of cholesterol to phospholipid substantiates the essential role of the phase transition in the process of liposome solubilization. When liposomes of the monotectic mixtures dimyristoyl and distearoyl phosphatidylcholine or dilauroyl and distearoyl phosphatidylcholine were incubated with plasma at temperatures in between those at which the constituent lipids undergo a phase change in the mixture, the liposomes were slowly dissolved. Under those conditions a selective removal of the lipids in the liquid-crystalline phase was observed. It is concluded that for the plasma-induced dissolution of unsonicated liposomes, which is most probably achieved by interaction with (apo)lipoproteins, the presence of phase boundaries is required in much the same way as was first reported for phospholipases by Op den Kamp, J.A.F., de Gier, J. and Van Deenen, L.L.M. (1974) Biochim. Biophys. Acta 345, 253--256).


Physical studies of cell surface and cell membrane structure. Deuterium nuclear magnetic resonance investigation of deuterium-labelled N-hexadeconoylgalactosylceramides (cerebrosides).

1. Deuterium Fourier transform nuclear magnetic resonance spectra of a series of N-palmitoylgalactosylceramides (cerebrosides) specifically labelled with deuterium at one of positions 2', 6', 10' and 16' of the acyl chain, or in the C-6 hydroxymethyl group of the galactose residue, have been obtained using a spin-echo technique at 34.1 MHz with a homebuilt superconducting magnet spectrometer. 2. The effects of temperature and cholesterol on the deuterium spectra have been investigated. The results indicate, when compared at the same reduced temperature, that the hydrocarbon chain organization in the liquid crystalline phase of palmitoylgalactosylceramide is essentially identical to that seen in similar chain length glycerophospholipids. In particular, two sets of quadrupole splittings are seen for a 2'-labelled N-palmitoylgalactosylceramide, indicating non-equivalent deuterons as noted previously for phospholipids. 3. Two sets of quadrupole splittings are observed for the headgroup C-6-labelled N-palmitoylgalactosylceramide. It is proposed that these signals arise from the enantiomeric R and S lipids, and that motion of the hydroxymethyl group is slow (greater than 10(-5) S). These results suggest the presence of a hydrogen bond network in the polar headgroup region. 4. The effects of cholesterol on the deuterium spectra of N-palmitoylgalactosylceramide-labelled as C2H3 in the terminal methyl group, at 1:1 mol ratios and in excess water below the crystal to liquid-crystal phase transition temperature (Tc) of the pure lipid (82 degrees C), are different to the effects seen with the phosphatidylcholine-cholesterol system. The spectra below Tc are characterised by two overlapping powder patterns, one with a quadrupole splitting of approx. 6 kHz (fluid liquid-crystalline phase) and one with a quadrupole splitting of about 20--25 kHz (crystal or gel-state lipid). Exchange between these two environments is therefore slow, leading to the possibility of characterising the cerebroside-cholesterol phase diagram using deuterium nuclear magnetic resonance spectroscopy.


Retention of lipid asymmetry in membranes on polylysine-coated polyacrylamide beads.

Phosphatidylcholine-specific exchange protein from calf liver was used to study the asymmetry and transmembrane movement of phosphatidylcholine in rat erythrocyte membranes isolated on polylysine-coated beads. While confirming previously published results for sealed ghosts, we found that for membranes attached to beads, where the cytoplasmic surface is exposed, about 36% of the total phosphatidylcholine is readily available for exchange, while the remaining 64% is exchangeable at a much slower rate. This indicates that the relative transbilayer asymmetry of phosphatidylcholine is largely maintained when red cell membranes are isolated on beads. On the other hand, transmembrane movement of phosphatidylcholine is decreased in membranes attached to cationized beads: the half-time for equilibration of phosphatidylcholine between the two monolayers of the membrane is 8 h for membranes on beads, compared to 1.5 h for sealed ghosts. Our results indicate that polylysine-derivatized beads are a useful tool for studying asymmetric properties of biological membranes.


Physicochemical studies of the protein-lipid interactions in melittin-containing micelles.

Complexes of melittin with detergents and phospholipids have been characterized by fluorescence, circular dichroism, ultracentrifugation, quasi-elastic light scattering and 1H nuclear magnetic resonance (NMR) experiments. By ultracentrifugation and quasi-elastic light-scattering measurements it is shown that melittin forms stoichiometrically well-defined complexes with dodecylphosphocholine micelles consisting of one melittin molecule and approximately forty detergent molecules. Evidence from fluorescence, circular dichroism and 1H nuclear magnetic resonance experiments indicates that the conformation of melittin bound to micelles of various detergents or of diheptanoyl phosphatidylcholine is largely independent of the type of lipid and furthermore appears to be quite closely related to the conformation of melittin bound to phosphatidylcholine bilayers. 1H NMR is used to investigate the conformation of micelle-bound melittin in more detail and to compare certain aspects of the melittin conformation in the micelles with the spatial structures of monomeric and self-aggregated tetrameric melittin in aqueous solution. The experience gained with this system demonstrates that high resolution NMR of complexes of membrane proteins with micelles provides a viable method for conformational studies of membrane proteins.


Purification and partial characterization of a procaryotic glycoprotein from the plasma membrane of Thermoplasma acidophilum.

The obligate, thermophilic, acidophilic mycoplasma, Thermoplasma acidophilum, grows optimally at 56 degrees C and pH 2.0. Its plasma membrane possessed 21--22 protein bands that were resolved by polyacrylamide gel electrophoresis. One major membrane protein, molecular weight 152 000, which stained for carbohydrate with periodic acid-Schiff reagent, accounted for 32% (w/w) of the total membrane proteins. It was isolated and further purified by concanavalin A affinity chromatography. The carbohydrate content amounted to less than 10% (w/w) compared to that of the entire glycoprotein. The carbohydrate moiety consisted mainly of mannose residues with branched alpha 1 leads to 2 linkages at the non-reducing ends of the glycopeptide as determined by permethylation followed by gas chromatography-mass spectrometry analysis. The reducing end was an N-glycosidic linkage between asparagine and N-acetylglucosamine. The amino acid composition of this glycoprotein showed 62 mol% hydrophobic residues, while the acidic amino acid content contributed 9 mol% more than that of the basic amino acids. The existence of membrane glycoproteins in the procaryotic, wall-less T. acidophilum may provide a protective coat for the plasma membrane. The stereochemistry and the conformation of the carbohydrate chains, in conjunciton with water turgor, may contribute to the rigidity of the membrane and the cation binding.


Biochemical and ultrastructural study of the disruption of blood platelets by streptolysin O.

The membrane-damaging protein toxin, streptolysin O, proved highly lytic on human, guinea-pig and rabbit platelets. About 15 molecules of toxin were sufficient to lyse one cell. Platelet disruption was assessed by electron microscopy, clearing of cell suspensions and assay of lactate dehydrogenase, serotonin, monoamine oxidase and glutathione peroxidase released in the extracellular fluid. This egress reflected the damage of both plasmic and organelle membranes. A quantitative study of lactate dehydrogenase and serotonin liberation taken as respective markers of the cytosol and dense bodies was undertaken as a function of toxin concentration. No platelet aggregation or shape change was elicited by streptolysin O. The ghosts resulting from platelet lysis retained properties of the native membrane such as aggregability and serotonin uptake. Dense bodies were easily separated after gentle disruption of the plasmic membrane by small amounts of toxin. Platelet lysis by streptolysin O proved a useful procedure for the determination of protein content, enzyme activities and serotonin assay on the same lysate in contrast to usual methods.


Red cell membrane in hemolytic disease. Studies on variables affecting electrophoretic analysis.

Significant alterations in the spectrin: band 3 and band 4.1a: band 4.1b ratios and an occasional decrease in the peak height of band 4.2 with respect to band 4.1 were found in electrophoretic patterns of red cell membranes from patients with hereditary xerocytosis. Electrophoretic comparison of whole cell, cytoplasm and membrane polypeptides implied that atypical partitioning at hemolysis could account for some, but not all, of the alterations seen in membrane patterns of xerocytes. A decrease in band 4.2 peak height as well as a variation in the profile of band 3 were produced in controls by specific manipulations of the electrophoresis protocol. Metabolic depletion of normal cells produced the type of alterations in bands 3 and 4.1 found in xerocyte membranes, whereas Heinz body production, addition of calcium to the hemolysis buffer and incubation of membranes in detergent under conditions designed to promote proteolysis did not. The presence of a higher peak height of band 4.1b with respect to that of band 4.1a in membranes of patients with various other red cell disorders correlated with an increase in the percentage of reticulocytes in peripheral circulation. The appearance of both band 3 and 4.1 abnormalities in the patterns of control cells which had been enriched in young cells by density gradient centrifugation suggested that these alterations in hemolytic disease are related to the predominance of young cells in the population.


Action of surface-active substances on biological membranes. IV. Hemolytic and membrane-perturbing action of homologous series of beta-D-glucopyranosyl-1-alkylphosphates.

The hemolytic action of a homologous series of beta-D-glucopyranosyl-1-alkylphosphates on human erythrocytes has been examined. The agent's affinity for the red cell membrane and the mean number of the agent's molecules which, upon interaction with an erythrocyte, make it undergo hemolysis have been measured. The contribution of the head group and that of a CH2 group of the surfactants to the free energy of the agents' binding to the cell membrane have been estimated. The effect of the surfactants on the red cell volume and the lytic concentrations of the agents have been measured. The contribution of a CH2 group to the free energy of the interaction of the amphiphiles embedded in the membrane bilayer with their environment has been evaluated and is proposed to be used as a measure of the membrane matrix stability.


An X-ray study of the cytoplasmic membranes of two gram-positive bacteria.

X-ray diffraction diagrams of partially disordered one-dimensional lattices of isolated bacterial cytoplasmic membranes are described and they provide a basis for suggesting possible molecular structures of bacterial membranes. Biochemical and electron microscope evidence points towards a lipid bilayer with a high degree of fluidity. The protein molecules are in a disordered configuration in the membrane.


Effects of changes in sodium electrochemical potential gradient on p-aminohippurate transport in newt kidney.

1. The relation between p-aminohippurate uptake and the electrochemical potential gradient of Na+ (delta muNa+) across the peritubular membrane was examined in newt (Triturus pyrrhogaster) kidney. The delta muNa+ was modified by changing cellular Na+ concentration and/or lowering the electrical potential difference across the peritubular membrane (peritubular membrane potential) 2. Elevation of external K+ concentration or addition of alanine at 40 mM to the medium decreased the delta muNa+ mainly through the depolarization of the cells. Addition of 1 mM ouabain resulted in a decrease in the peritubular membrane potential and increase in cellular Na+ concentration, thus decrease in the delta muNa+. 3. p-Aminohippurate uptake decreased in proportion to the decrease in the delta muNa+ under all experimental conditions, indicating that the maintenance of the delta muNa+ is required for p-aminohippurate transport. 4. All three different experimental conditions, high medium K+ concentration, 40 mM alanine or 1 mM ouabain, increased the apparent Michaelis constant, Kt, without affecting the maximal uptake rate, V, for p-aminohippurate. These results suggests that the delta muNa+, largely the peritubular membrane potential, may affect the association and/or dissociation of p-aminohippurate and Na+ at both interfaces of the peritubular membrane of the proximal tubular cells.


Inhibition of 14C-labeled amino acid incorporation into protein by particles and surface-active agents in human granulocytes.

Contact of human polymorphonuclear leukocytes with latex or zymosan particles resulted in inhibition of incorporation of 14C-labeled amino acids into protein in the phagocytosing cells. The surface-active agents, concanavalin A and digitonin, exerted the same inhibitory activity on 14C-labeled amino acid incorporation into protein in human granulocytes. The inhibition of 14C label incorporation was dose dependent and the half-maximal effect per 10(7) cells was found at 0.8 mg of latex (0.79 micrometer in diameter), 0.25 mg of serum-coated zymosan and 35 microgram of digitonin/ml of the incubation medium. Amino acid transport (14C-labeled amino acids or amino[14C]isobutyrate) was the same in the resting and phagocytosing granulocytes. These results suggest that the incorporation of 14C-labeled amino acids into protein in human granulocytes is in some way connected to alteration in the cell plasma membrane.


Biosynthesis of ribosomal proteins by poly(A)-containing mRNAs from rat liver in a wheat germ cell-free system and sizes of mRNAs coding ribosomal proteins.

(1) Poly(A)-containing mRNAs from total polysomal RNA of regenerating rat liver were incubated with [3H]leucine in a wheat germ cell-free system. Ribosomal proteins were purified as described previously [1], and with two-dimensional gel electrophoresis. The proteins on the gel except for less basic protein had appreciable radioactivity, whereas the surrounding areas had very low radioactivity. Acetic acid-soluble proteins labeled in this system were subjected to three-dimensional gel electrophoresis [2]. Except for L1 and L2 proteins, each of the ribosomal proteins, including less basic ones, showed a major radioactive peak coinciding with the protein band on SDS gel. Thus, the wheat germ cell-free system completely translates almost all mRNAs for individual ribosomal proteins. Equimolar amounts of almost all ribosomal proteins were synthesized in the presence of the saturating concentration of mRNAs. (2) Free polysomes from regenerating rat liver were fractionated into three sizes. Each class of polysomes was incubated with [3H]leucine. Ribosomal proteins with molecular weights of 40 000 to 21 000 were mainly synthesized by Fraction B (5-14 monomeric ribosomes), L1 and L2 [2] with 60 000 and 54 000, by Fraction C (greater than 15 monomeric ribosomes) and B, and ribosomal proteins smaller than 20 000 by Fractions A (less than pentamer) and B. (3) mRNAs from rat liver total polysomes were fractionated into seven classes by size and each was translated in the wheat germ extract. Ribosomal proteins with molecular weights of 54 000 to 30 000 were mainly synthesized by mRNAs of 12 to 14.5 S, ribosomal proteins of 35 000 to 22 000 by those of 9.5 to 12 S, ribosomal proteins of 22 000 to 13 000 by those of 7 to 9.5 S, and smaller ribosomal proteins by those smaller than 7 S. These results indicate that individual ribosomal proteins are synthesized by monocistronic mRNAs, the lengths of which are proportional to the molecular weights of the corresponding ribosomal proteins.


Xeroderma pigmentosum variants have a slow recovery of DNA synthesis after irradiation with ultraviolet light.

Human cells (normal and xeroderma pigmentosum variant) irradiated with ultraviolet light and pulse-labelled with [3H]thymidine underwent transient decline and recovery of molecular weights of newly synthesized DNA and rates of [3H]thymidine incorporation. The ability to synthesize normal-sized DNA recovered more rapidly in both cell types than thymidine incorporation. During recovery cells steadily increased in their ability to replicate normal-sized DNA on damaged templates. The molecular weight versus time curves fitted exponential functions with similar rate constants in normal and heterozygous xeroderma pigmentosum cells, but with a slower rate in two xeroderma pigmentosum variant cell lines. Caffeine added during the post-irradiation period eliminated the recovery of molecular weights in xeroderma pigmentosum variant but not in normal cells. The recovery of the ability to synthesize normal-sized DNA represents a combination of a number of cellular regulatory processes, some of which are constitutive, and one of which is altered in the xeroderma pigmentosum variant such that recovery becomes slow and caffeine sensitive.


An assay system for factors involved in mammalian DNA replication.

An assay for cellular factors stimulating DNA synthesis by partially lysed CHO cells is presented. The assay is based on the observation that in highly lysed cells, DNA synthesis, as determined by [3H]dTTP incorporation, was only 2-5% of that in gently lysed cells, and that this low level of DNA synthesis could be increased by a factor of approx. 50 by the addition of CHO cell extract (i.e. supernatant of a cell homogenate subjected to high-speed centrifugation). Highly lysed cells were obtained by treatment with 0.1% Brij-58 and 240 mM KCl, while for the preparation of gently lysed cells, 0.01% Brij-58 and 80 mM KCl were used. Incorporation of [3H]dTTP reflected DNA synthesis qualitatively similar to that in intact cells. It was semiconservative, and no repair synthesis was detected unless cells were irradiated with ultraviolet light prior to parital lysis. DNA molecules of 4 S were synthesized and converted to DNA of more than 25 S via 6-12-S intermediates. DNA synthesis was restricted to nuclei from cells in S phase, and cell extract did not induce DNA synthesis in nuclei from cells in G1 phase. Stimulation of DNA synthesis by cell extract was concentration-dependent. Cell extract activity was recovered to more than 50% after (NH4)2SO4 precipitation. Heat-inactivation experiments suggested that cell extract contained at least tow factors timulating DNA replication. This system may, therefore, be used for the purification and characterization of factors participating in DNA replication of mammalian cells.


Sequential changes in DNA polymerases alpha and beta during diethylnitrosamine-induced carcinogenesis.

It has often been suggested that the high molecular weight DNA polymerase alpha of eukaryotes plays a role in de novo replication of DNA, while the low molecular weight polymerase beta is involved in repair replication. Previous studies have shown that when diethylnitrosamine is fed in the diet to rats it causes after a few weeks an increase in de novo replication of DNA, which then returns to normal values. In contrast, repair replication may be expected to continue throughout the feeding period. Study of DNA polymerase activity in livers of animals during carcinogenesis showed that an increase in polymerase alpha occurred at the time of increased de novo replication, while there was a gradual increase in polymerase beta during the time diethylnitrosamine was present in the diet. When diethylnitrosamine treatment was stopped, there was a rapid drop in polymerase beta activity. These results support the view that the polymerase alpha is involved in DNA replication, that the polymerase beta functions in repair replication, and that the beta enzyme can be induced by chronic damage to DNA.


Isolation of a stimulatory factor for nuclear DNA replication.

Aqueous extracts of isolated nuclei and intact plasmodia of Physarum contain a heat-stable stimulator of nuclear DNA replication. The stimulatory factor is present throughout the mitotic cycle, and its activity is unaffected by prior exposure of plasmodia to cycloheximide. The stimulatory substance has been partially purified by heat treatment, precipitation with ethanol, chromatography on DEAE cellulose, and gel filtration. The purified material contains both carbohydrate and protein, and exhibits a molecular weight of about 30 000. The active substance increases the rate and overall extent of DNA replication in S-phase nuclei, but does not trigger the initiation of DNA synthesis in nuclei isolated from G2-phase plasmodia. The stimulatory material contains little or no deoxyribonuclease or DNA polymerase activity, and it does not affect DNA polymerase activity assayed using a purified DNA template.


Immunochemical detection of N6-methyladenine in DNA.

Antibodies to N6-methyladenosine were produced in rabbit by means of immunization with N6-methyladenosine coupled to bovine serum albumin via periodate oxidation. Cross-reacting antibodies were removed by bovine serum albumin-Sepharose and appropriate nucleoside-human serum albumin absorbents. Nucleoside-specific antibodies were isolated by affinity chromatography of N6-methyladenosine-human serum albumin-Sepharose. The specificity of the purified antibodies has been demonstrated by complement fixation inhibition analyses using nucleoside analogues as inhibition than dAMP. Anti-N6-methyladenosine was used to detect N6-methyladenine in denatured DNAs from various sources by complement fixation. Practically no complement fixation has been found with DNAs containing no N6-methyladenine, such as calf thymus, salmon sperm, Micrococcus radiodurans, Streptomyces chrysomallus and Streptomyces hygroscopicus, whereas a weak reactivity occurred in the case of Bacillus subtilis DNA and Sarcina maxima DNA. For DNA from Proteus mirabilis, Escherichia coli K-12, E. coli B, E. coli WF+, lambda, T2 phages quantitative differences in the immunochemical reactivity were observed, which only partially correlate with the N6-methyladenine content of the DNAs. Other factors, influencing the accessibility of N6-methyladenine to the antibody-combining site have to be taken into consideration.


Levels of aminoacyl-tRNA synthetases, tRNA nucleotidyltransferase and ATP in germinating lupin seeds.

Transfer RNAs in dry lupin seeds are aminoacylated to a low extent (Kedzierski, W. and Pawełkiewicz, J. (1977) Phytochemistry 16, 503-504) and are partly degraded at the acceptor terminus (Dziegielewski, T. and Pawełkiewicz, J. (1977) Bull. Acad. Polon. Sci. Ser. Biol. 7, 4oo-435). Increase in the levels of tRNA aminoacylation and disappearance of defective tRNA molecules during seed germination are not accompanied by significant changes in the levels of phenylalanyl-, arginyl-, valyl-tRNA synthetases and tRNA nucleotidyltransferase. Additionally, no inhibitor of aminoacylation of valine tRNA has been detected in dry seeds. However, dry seeds contain very low ATP amounts, which increase dramatically during germination. The above results suggest that a very low ATP level is a factor limiting the aminoacylation and reparation of tRNA molecules at early stages of seed germination.


Low molecular weight RNAs as components of nuclear ribonucleoprotein particles containing heterogeneous nuclear RNA.

70-130 S polyparticles as well as 38 S monoparticles were isolated from rat liver nuclei and analyzed in respect to their RNA components by microgel polyacrylamide electrophoresis in formamide. In addition to the high molecular weight polydisperse hnRNA of polyparticles several low molecular weight RNAs (snRNA) were detected. There are at least six distinct snRNA species in polyparticles. Except for one species, which is missing, 38 S monoparticles showed a similar snRNA pattern. From densitometer tracings of microgels the snRNAs were estimated to represent about 11% of the total polyparticle RNA. The number of nucleotides for the various snRNAs were determined from a plot of relative electrophoretic mobility versus log number of nucleotides. The possibility that the snRNAs are degradation products of the hnRNA was excluded on the basis of the following findings. (1) The snRNA pattern was similar in mono- and polyparticles. (2) Whereas the hnRNA of polyparticles incubated at 37 degrees C was extensively degraded, the snRNA did not show a corresponding increase. (3) After a 30 min pulse with [3H]orotate the hn RNA was readily labeled; none of the snRNAs, however, incorporated radioactivity. The snRNAs were still found after treatment of polyparticles with 2 M NaCl excluding contamination by nucleoplasm.


Polyribosome size analysis. Measurement of number-average polyribosome sizes.

The analysis of translational efficiencies of specific mRNAs requires a determination of the polyribosome size. The appropriate value to use in such calculations is the number-average size. A method is described for accurately measuring the number-average size of total and of specific protein synthesizing polyribosomes using isokinetic sucrose density gradients and 125I-labeled antibodies. By this method, we demonstrated that albumin synthesizing polyribosomes from a serum albumin secreting mouse hepatoma cell line exist over a broad range from trimers to 20-mers (mean 6-10). The specificity of antibody interaction with polyribosomes was demonstrated using cells not synthesizing mouse serum albumin, and by demonstrating that 125I-anti ovalbumin does not bind to mouse hepatoma polyribosomes. Treatment of the mouse hepatoma cells with 1 MUM cycloheximide shifted practically all of the monomers into polyribosomes resulting in an increase in the number-average size of the albumin synthesizing polyribosomes. Cycloheximide treatment, however, did not eliminate the size heterogeneity in the albumin synthesizing polyribosomes.


The occurrence of uridine diphosphate N-acetylgalactosamine 6-sulfate in quail egg white and characteristic distribution of sulfated sugar nucleotides in different avian eggs.

A sulfated sugar nucleotide has been isolated from quail egg white, and accounts for nearly 80% of the total sugar nucleotides found in the egg white. Evidence is presented that this nucleotide is uridine diphosphate N-acetylgalactosamine 6-sulfate, an isomer of the 4-sulfated derivative of uridine diphosphate N-acetylgalactosamine previously found in chicken egg white. Further studies on the distribution of sulfated sugar nucleotides in egg white of various birds (chicken, quail, pheasant, peafowl, turkey, goose, and duck) demonstrate that each species has a characteristic composition, differing from one another regarding the relative amounts of 4-sulfated, 6-sulfated, and 4,6-bissulfated derivatives of uridine diphosphate N-acetylgalactosamine.


Association of poly(ADP-rib) synthesis with cessation of DNA synthesis and DNA fragmentation.

CHO cells and cs-4-D3 cells were used to investigate the association between poly(ADP-rib) synthesis and the cessation of DNA synthesis and DNA fragmentation. The cs4-D3 cells are cold-sensitive DNA synthesis arrest mutants of CHO cells. Upon incubation at 33 degrees C, DNA synthesis in the cs4-D3 cells stops and the cells enter a prolonged G1 or G0 phase. The events that occurred when cs4 cells were incubated at 33 degrees C were similar to those that occurred when wild-type CHO cells grew to high density. (1) In both cases, DNA synthesis and cell growth stopped. (2) The NAD+ concentration/cell was 20-25% lower in growth-arrested cells than in logarithmically growing cells. (3) Poly(ADP-rib) synthesis was 3-4 fold higher in growth-arrested cells than in logarithmically growing cells. (4) The growth-inhibited cells developed DNA strand breaks which resulted in large percentages of their DNA appearing in the low molecular weight range of alkaline sucrose gradients. (5) Both the increased rate of poly(ADP-rib) synthesis and the development of DNA strand breaks appears to be characteristic of the G1 phase of the cell cycle. (6) When growth-inhibited cells were restored to conditions favorable for DNA synthesis and cell growth, the DNA strand breaks were repaired. (7) Prolonged incubation under growth-restrictive conditions resulted in the accumulation of more DNA strand breaks than the cells could repair. This was followed by cell death when the cells were restored to conditions favorable for cell growth.


Proton magnetic resonance studies on [methionine]-enkephalin and beta-endorphin in aqueous solution.

Proton magnetic resonance studies of [Met5]-enkephalin (lipotropin 61-65) in aqueous solution indicate a conformational preference for the pentapeptide backbone. The structural differences between [Met5]-enkephalin and other, more flexible peptides have been investigated using paramagnetic probe techniques. An outline structure for beta-endorphin (lipotropin 61-91) in aqueous solution is obtained from binding studies using Gd(III) as a relaxation probe.


Intrachain disulfide bridges of bovine seminal ribonuclease.

The pairing of the four intrachain disulfide bonds of bovine seminal ribonuclease, a dimeric protein isolated from bovine seminal plasma, has been established by the isolation and characterization of the cystine peptides obtained from a thermolytic-tryptic hydrolysate of the protein. These disulfide bonds involve eight half-cystine residues located in the protein subunit chain at sequence positions identical with those of the eight half-cystine residues of the strictly homologous chain of bovine pancreatic ribonuclease. The results reported show that these eight 'homologous' half-cystine residues pair in seminal ribonuclease exactly as they do in pancreatic ribonuclease. They also indirectly confirm that the remaining two half-cystine residues present in each chain of the seminal enzyme are involved in intersubunit bonds.


Studies on factor VIII-related protein. III. Size distribution and carbohydrate content of human and bovine factor VIII.

Human and bovine factor VIII were isolated from cryoprecipitate of fresh frozen plasma by gel filtration on Sepharose CL-2B. The elution diagrams and SDS-agarose electrophoretic analysis of eluted fractions show no significant differences in the size-distribution of factor VIII aggregates between the two species. Agarose gels were stained for carbohydrate by two methods: (1) the dansyl hydrazine reaction following oxidation with periodic acid and (2) staining with fluorescein-labeled concanavalin A. Results of both procedures indicate that in human factor VIII neither the size distribution nor its ristocetin cofactor activity are related to carbohydrate content. Bovine factor VIII contains slightly less sugar than the human preparation as judged from the relative dansyl hydrazine staining intensities. In contrast to human factor VIII, the binding affinity for concanavalin A of bovine factor VIII was gradually decreased with increasing aggregate size. This finding suggests an impaired accessibility of reactive sugar residues in large aggregates of bovine factor VIII.


Characterization of ferritin from human placenta. Implications for analysis of tissue specificity and microheterogeneity of ferritins.

Mammalian ferritins can be resolved into multiple components by isoelectric focusing, and each tissue contains a characteristic subset of isoferritins. Ferritin isolated from human liver was compared to acidic ferritin isolated from mid-gestational human placenta to define a structural basis for ferritin heterogeneity. Placenta ferritin contained several major bands with isoelectric points in the range of pI = 4.7-5.0 which were more acidic than the predominant isoferritins of human liver. Ferritin from each tissue was resistant to denaturation by 10 M urea and appeared to be identical by electron microscopy. Circular dichroism measurements revealed that placenta ferritin had substantially less ordered secondary structure than liver ferritin. Both types of ferritin contained only two subunits when analyzed by electrophoresis in sodium dodecyl sulfate gels, but isoelectric focusing of dissociated subunits in urea revealed 6-7 different components. In this system, placenta ferritin was enriched in the more acidic subunits and it completely lacked the most basic subunits noted in liver ferritin; placental ferritin had no unique components. Differences in isoelectric points among assembled ferritins from these two tissues appear to result from different proportions of these acidic and basic subunits.


Purification of the insulin receptor from human placental membranes.

Insulin receptors were purified from human placental microsomal membranes by solubilisation with Triton X-100 followed by Sepharose 6B chromatography, phosphate gradient elution from hydroxyapatite and affinity chromatography on concanavalin A-Sepharose. 2000-fold purification was achieved with 63% overall recovery. The purified receptor gave a single band on 3.75% polyacrylamide (0.1% Triton X-100) gel electrophoresis. On sodium dodecyl sulphate-polyacrylamide gel electrophoresis there was a major band at 75,000 and a minor band at 80,000 daltons. The purified receptor rechromatographed on Sepharose 6B with an apparent molecular weight of 300,000.


Purification and characterization of rat angiotensinogen.

1. Angiotensinogen (renin substrate) was purified from plasma of nephrectomized rats by a four step procedure using ammonium sulfate fractionation, chromatography on Blue Sepharose CL-6B and SP-Sephadex C-50, and gel filtration on Sephadex G-150. 2. The final preparation had a specific concentration of 9.3 microgram angiotensin I/mg (mean of six separate runs). The best preparation so far obtained contains 14.6 microgram angiotensin I/mg protein, which represents a purity of 62%. 3. By sodium dodecyl sulfate disc electrophoresis an apparent molecular weight of 56,400, and by isoelectric focusing an isoelectric point of 4.85 has been determined. These properties of rat angiotensinogen are similar to those reported for human angiotensinogen.


Purification and characterization of a glycopeptide derived from Phaseolus vulgaris leukoagglutinating phytohemagglutinin.

A glycopeptide obtained from tryptic digestion of citraconylated leukoagglutinating (L-subunit) phytohemagglutinin was isolated and purified. The composition of its ten amino acid and seven hexose residues, when compared to a previous analysis of the NH2-terminal amino acid sequence, indicated that the glycopeptide was derived from residues 11 through 20, with the oligosaccharide unit linked N-glycosidically to asparagine at the twelfth position. Structural studies involving a combination of alpha-mannosidase digestion, periodate oxidation, and permethylation, in conjunction with computerized mass-spectrometric and masschromatographic techniques, suggested a branched sequence of five mannose and two glucosamine residues similar to that found in many mammalian glycoproteins.


Purification, characterization and identification of an agglutinin in human serum.

A serum protein named agglutinin is able to induce mitochondria to agglutinate. The protein has been purified from human serum by chromatography on DE-52. Sephadex G-200 and immunoglobulin-Sepharose 4B columns. Agglutinin is a glycoprotein that migrates electrophoretically as a gamma-globulin. Its molecular weight was determined to be 50,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Monospecific antiserum prepared against the agglutinin was found to be identical with anti-beta 2-glycoprotein I and agglutinating activity could be adsorbed on anti-beta 2-glycoprotein I-Sepharose 4B columns. Thus, the agglutinin has been identified as beta 2-glycoprotein I. The reaction between mitochondria and agglutinin shows positive cooperativity, which is independent on the stage of purification of agglutinin. The agglutinating activity could be diminished (inhibited) by acidic non-soluble lipids such as oleic acid, phosphatidic acid, phosphatidyl serine and phosphatidyl inositol.


Kinetics of the in vitro gelation of a sickling haemoglobin from Hog deer (Axis porcinus).

In order to distinguish the molecular mechanisms involved in the polymerisation of animal and human sickling haemoglobins, the gelation properties of hog deer (Axis porcinus) haemoglobin have been studied. Continuous monitoring of viscosity and minimum gelling concentration measurements of hog deer haemolysates were made over a range of pH, temperature, ionic strengths and in the presence of urea and tris(hydroxymethyl)methylamine. The inhibition of gelling caused by lowering the pH or increasing ionic strength and the abolition of the reversible endothermic nature of gelation by urea suggest that electrostatic interactions predominate in polymerisation but that weak hydrogen and hydrophobic bonds may also be present. Kinetic viscosity data demonstrated a pre-gelation lag phase, dependent on ahigh power of the haemoglobin concentration, similar to the nucleation of human haemoglobin S (HbS). The results indicate similarities in the kinetics of hog deer polymerisation with those of HbS, but major differences in the type of intermolecular attractions involved.


Earthworm (Pheretima communissima and Pheretima hilgendorfi) hemoglobins: giant assemblies and subunits.

The molecular architecture of hemoglobins and their subunits of the earthworms Pheretima communissima and Pheretima hilgendorfi was investigated. In both species, their s0.20,w of 60.8 S and D020,w of 1.80 . 10(-7) cm2 . s-1 corresponded to a molecular weight of 3.07 . 10(6). From electron microscopic observations, the overall structure of the hemoglobins was shown to be two superimposed hexagonal discs, each composed of six-membered constituents. Sodium dodecyl sulfate polyacrylamide gel electrophoresis of both hemoglobins revealed the presence of five species of subunits with molecular weights of 13,000-14,000 (subunit 1), 27,000-28,000 (subunit 2), 30,000-31,000 (subunit 3), 33,000-34,000 (subunit 4) and approx. 52,000 (subunit 5), respectively, and the molar ratio of these subunits 1:2, 3, 4:5 was 2:3:3. If we consider this set of the subunits 1 to 5 as one unit, the molecular weight of this unit should be 2.7-2.8 . 10(5). This one unit, therefore, should be considered to represent one-twelfth the whole molecule with molecular weight of 3.07 . 10(6).


Relative hydrophobicity of human serum proteins as measured by partition in aqueous two-polymer phase systems.

The effect of the ionic strength on the partition of a number of human serum proteins in the buffered isotonic ficoll-dextran phase systems was examined. The relative hydrophobicity of the protein was estimated.


Structure of a Gla-containing dipeptide. The crystal structure of (+/-)-N-carbobenzoxy-(gamma,gamma'-DI-tertbutyl)-gamma-carboxyglutamylglycine ethyl ester.

The crystal and molecular structure of a dipeptide containing a blocked gamma-carboxyglutamyl (Gla) residue is presented. Two intermolecular hydrogen bonds link the amides with carbonyl groups in the dipeptide backbone, but the protected gamma-carboxy groups on the modified glutamic acid are not hydrogen bonded.


Occurrence of Leu-Lys-bradykinin and histidine-rich peptide in high-molecular-weight kininogen isolated from horse plasma.

On incubation of purified horse plasma high-molecular-weight kininogen with purified plasma kallikrein, three new peptides, named fragment 1.2, fragment 1 and fragment 2, were released, in addition to the vasopeptide, bradykinin. Fragment 2 contained an extremely high level of histidine, in which eleven residues out of the total 48 residues were characterized. Thus the result proves the existence of the histidine-rich region in horse high-molecular-weight kininogen, which is similar to the region previously identified in bovine high-molecular-weight kininogen. Moreover, we have identified a new kinin derivative, Leu-Lys-bradykinin, in horse high-molecular-weight kininogen.


Change in binding properties of folate-binding protein in cow's whey due to removal of a cofactor during affinity chromatographic purification.

The folate-binding protein in cow's whey was purified by affinity chromatography on folate or methotrexate-AH-Sepharose 4B. A change in basic binding properties of the protein occurred probably due to the fact that material removed during affinity chromatography contains a cofactor of great importance to folate binding.


Purification and properties of human erythrocyte delta-amino-levulinic acid dehydratase (EC 4-2-1-24).

Human delta-aminolevulinic acid dehydratase (ALA-D) was purified 9 000-fold by salt precipitation, ion-exchange chromatography and gel filtration. These methods resulted into an electrophoretically and immunologically pure protein. The optimum pH of the enzyme is 6.6 and its Km with ALA : 4.8 X 10(-4) M. The enzymatic activity was increased by thiol-containing substances, such as dithiothreitol (DTT), which protect the -SH groups of the protein. Zinc, a portion of the enzyme molecule, was partly lost during the purification procedure; its addition enhances the enzymatic activity. Determination of molecular weights and electron microscopy study are in favor of an octameric structure.


The apoprotein fraction of the bile lipoprotein complex: isolation, partial characterization and phospholipid binding properties.

A bile apoprotein fraction (Apo BLC) was isolated by preparative isoelectric focusing (I.E.F.) from the detergent-free form of the bile lipoprotein complex (BLC). Analytical I.E.F. of Apo BLC yields a characteristic and reproducible pattern of two narrow acidic bands (pI 4,8-5,0). This apoprotein presents a strong tendency to undergo self-aggregation in aqueous buffer. A low molecular weight constituent of Apo BLC has been isolated after gel filtration, its mean Mw is estimated by SDS-PAGE at 7,500 daltons. The binding capacity of Apo BLC for phospholipids was investigated on dimyristoylphosphatidylcholine liposomes by gel filtration and zone electrophoresis. The resulting structures, larger than the original single-shelled vesicles, acquire and anodic electrophoretic mobility. Apo BLC has a weaker affinity for lysophosphatidylcholines: these phospholipids decrease the degree of aggregation of the apoprotein. These studies contribute additional data concerning the high affinity of Apo BLC for phosphatidylcholines, which are the major phospholipid constituents of bile. The discussion deals with the fact that association of Apo BLC with bile phosphatidylcholines may present some implications in the pathogeny of LpX and in the process of intestinal fat absorption.


[Rat liver phosphoglycerate kinase. I. Purification and kinetic properties in the biosynthesis of 1-3 diphosphoglycerate].

Phosphoglycerate kinase (MgATP 3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) has been isolated from rat liver with a purification ratio of 960 and a specific activity of 300 IU/mg of protein. The purity of the enzyme preparations was estimated by polyacrylamide gel electrophoresis. The molecular weight, determined by gel filtration is 42 000. The "subunit" size of phosphoglycerate kinase as determined by sodium dodecyl sulfate gel electrophoresis is 46 000, indicating that the enzyme is monomeric. The rate of the enzyme reaction as a function of the concentration of D-3-phosphoglycerate indicated the usual Michaelis Menten relationship. The rate of the enzyme reaction as a function of the concentration of MgATP2- did not fit the usual Michaelis Menten relationship: two distinct regions can be fitted with different straight lines and suggest the presence of two sites for the Mg ATP2-. This hypothesis seems to be confirmed by the study of the action of the free and complexed nucleotides.


[Equilibrium of thiamine diphosphate complexation with divalent ions--formation constants].

The divalent metal ion-thiamine diphosphate (TDP) complexes are of potential interest because such complexes are directly involved in enzymatic reactions. The binding of Ni2+ to TDP has been studied in order to gain some insight in the TDP-metal interactions which could help to understand the role of the metal ions in general in the enzymatic processes. A technique of pH titration is developped which brings definitive proof of metal-TDP bonding in aqueous solutions. The results indicate that the metal ion interacts both with the pyrophosphate group and the pyrimidine ring. The complexation decreases the pK for the phosphate ionization. The ring nitrogen is a some 12 times weaker ligand of Ni2+ than is pyrophosphate. The stability constants obtained for an ionic strength 0.5 in tetramethylammonium bromide are reported; the maximum of the apparent formation constant, between pH 4.5 and pH 7.5, occurs for pH 6.9. The poor solubility of nickel hydroxyde is responsible for increasing interaction in alcaline solution.


Late positive component correlates of verbal and visuospatial processing.

Bilateral VERs were elicited by letter stimuli requiring verbal processing and patterns requiring visuospatial processing in a go/no-go reaction time paradigm. The VERS were formed separately to the targets and non-targets of each stimulus type. No VER component displayed hemisphere asymmetries which varied in a task-dependent fashion. However P150 latency was found to be shorter, and N150 amplitude larger, in VERs recorded from the right hemisphere. This is considered to reflect the superiority of the right hemisphere for the early stages of the processing of visual input. P400 latency was shorter and its amplitude larger to the pattern stimuli. In addition, subjects' reaction times were shorter to these stimuli. These results were interpreted in evidence supporting the notion that the late positive component of the ER reflects processes of stimulus evaluation which are not necessarily correlated with those of response selection and execution.


Consistency of individual differences in non-specific electrodermal activity.

The study sought to determine whether, as implied by a trait interpretation of individual differences in non-specific electrodermal responding, the frequency of NSRs is a consistent characteristic of subjects across conditions. 40 male subjects completed two 1-hour sessions spaced at an interval of 2 weeks. In each session, three 12-min conditions were administered: relaxation, vigilance, and mental arithmetic. Continuous recordings were made throughout each condition of electrodermal and respiratory activity, and at the end of each condition subjects completed an Adjective Checklist (ACL). Counts were made of the total number of 10-sec intervals during each condition in which at least one NSR was recorded. Separate counts were made for large amplitude (equal to or greater than 1% of baseline) and for small amplitude (less than 1%) responses. Analyses of variance performed on the NSR data indicated that the major component of variance was that due to subjects, and that the interactions between subjects and conditions and subjects and sessions, which would contradict a trait interpretation, were negligible. Consistency in subject's responding was estimated by intraclass correlations across conditions and sessions. For NSR large, the coefficient was 0.58 and for NSR small 0.50. The data were interpreted as support for a trait interpretation of NSR frequency.


Autonomic classical conditioning as a function of awareness of stimulus contingencies.

Two groups of 32 college students were presented compound CSs (lights and tones presented simultaneously) during a classical conditioning paradigm. By means of a masking task and verbal instructions, a partially informed group was made aware of only the visual CS's contingency with the UCS, while a fully informed group was made aware of both the visual and auditory contingencies. Autonomic indices of conditioning (electrodermal responses, heart rate, and digital pulse volume) were later measured to the individual component CSs and to various compound CSs. It was found that: (1) the partially informed group exhibited conditioning exclusively to the visual CS+ and to compounds which included the visual CS+, while (2) the fully informed group exhibited conditioning to both visual and auditory CS+s. The results confirm the importance of awareness in human autonomic discrimination classical conditioning. It is suggested that human autonomic conditioning may be usefully conceptualized as an information processing task with the autonomic indices of conditioning reflecting central cognitive processes.


Interaction effects of extraversion and neuroticism on detection thresholds.

Detection thresholds for electrocutaneous stimulation were measured in young male subjects by two different methods - a method of limits and a forced-choice method - the latter assumed to be less influenced by decision processes. Relations between the threshold measures and scores in the EPI extraversion (E) and neuroticism (N) scales were studied. High N-low E ("unstable introvert") subjects had the lowest thresholds. The result is in line with the predictions made on the basis of Eysenck's personality theory.


Ultradian rhythms in alertness - a pupillometric study.

Pupillary size, pupillary light reflex and pupillary motility were measured every 15 min, for 10 continuous hours, in eight young adults. Time series constructed from the range of pupillary motility revealed 75 to 125 min rhythms; these were significantly out of phase with rhythms of similar periodicity in pupillary diameter and in the magnitude of the pupillary light reflex. There was no consistent correspondence between Stanford Sleepiness Scores and pupillary activity. It is suggested that the rhythms in pupillary activity reflect underlying rhythms in CNS arousal that also modulate perceptual, cognitive and electroencephalographic processes, a conclusion which supports Kleitman's BRAC model.


Physiological response patterns during "intake" and "rejection" tasks.

Two 2-min tasks, one requiring outer-directed attention (intake task), the other a mental arithmetic task (rejection task) were presented to 20 subjects. According to Lacey the distinguishing feature of the response to intake tasks are cardiac deceleration and blood pressure decreases. None of these responses was obtained. Some variables, however, were only affected by one of the tasks; the intake task led to a decreased hand (skin) blood flow and an increased hand vascular resistance, whereas a heart rate acceleration and increased forearm (muscle) blood flow and decreased forearm vascular resistance were observed during the rejection task. Blood pressure, respiration rate and skin conductance were similarly affected by the two tasks. Inconsistencies among previous studies of the response to the two types of tasks were discussed and found to be partially explained by methodological differences.


The patho-physiological basis for tests of viability in isolated perfused organs.

The causes and prevention of inadequate isolated organ perfusion are discussed. The consequences of inadequate perfusion are outlined and presented as a rationale for the choice of appropriate tests of viability. The routine application of some tests is advocated and the criteria used in the assessment of the results of such test considered.


Physiological ageing of red blood cells and changes in membrane carbohydrates.

Evidence is presented to indicate a generalized role for the terminal sialic acid residues of circulating erythrocytes. After reinjection into their donors, neuraminidase-treated human, rabbit, rat and dog erythrocytes were promptly removed from the circulation : intect erythrocytes, previously incubated under the same conditions but without neuraminidase, were removed after a significantly longer period. The neuraminidase-treated erythrocytes were cleared by the liver and in a little part by the spleen. Old and young human, rabbit, rat erythrocytes contained different quantities of stromal sialic acid, significantly lowered on the old cells. But the half-life of old intact rabbit erythrocytes is sigificantly shorter than that of neuraminidase-treated young erythrocytes with a similar minidase-treated young erythrocytes with a similar sialic acid content. Indeed sialic acid is not the only carbohydrate component of the membrane that is decreased during erythrocyte ageing, the others membranous sugars are decreased too. Theses changes in the carbohydrate moity could have a role in the clearance of the erythrocytes.


Suitability criteria for compartmental analysis of the plasma clearance curve of exogenous cholephils. 131I Rose Bengal fulfils these criteria.

Hepatic clearance of exogenous cholephils involves at least three transfers : uptake by the liver, reflux from hepatocytes into the plasma, and intracellular transport followed by active secretion in bile. Compartmental analysis may be applied to the plasma disappearance curve in order to calculate the fractional transfer rates. The 131I Rose Bengal (131I RB) fills the requirements of such an analysis, contrary to bromsulphalein and indocyanine green, which have been used until now. 131I RB does not undergo enterophepatic cycle and is not metabolized by the liver. It behaves, in the preparation used, like a homogeneous substance, remains stable in the serum as long as 25 hours after its injection and may be bound completely with human albumin in vitro prior to administration : under these conditions, the material which is cleared by the liver is homogeneous. The use of tracer instead of loading doses, avoid any toxic effects on transport systems and results in plasma and liver concentrations which are negligeable in comparison with the Km of membrane transport, and are compatible with first order kinetics.


Action of middle molecules from chronic renal insufficiency treated by hemodialysis on mitochondrial respiration.

"Middle molecules" are isolated both from the urine of normal subjects and from the blood of patients suffering from chronic renal insufficiency treated by hemodialysis. Their action is tested on mitochondrial respiratory activity. Some of these molecular species (fraction 7 f and 7 g) are without any effect. On the other hand the 7 c middle molecules fraction gives a loss of respiratory control due to a decrease of respiratory activity in state 3. A diminution of the P/O ratio is noted and so the available ATP pool is decreased. This action can be compared to the fact that 7 c plasmatic fraction concentration seems to be connected with the patients clinical condition.


Teratogenic effects of lead in the mouse.

Female mice of the C57B1 strain were mated and given from the first day of the pregnancy a normal diet, containing 1.1% of calcium, or a calcium-deficient one, containing 0.2% of calcium. Animals of the 2 groups were injected intra-peritoneally with 15 or 35 mg of lead acetate/kg at different times of the fetal organogenesis (8th, 9th, 10th or 12th day of the pregnancy). In the normal diet group, injection of lead increases the postimplantation mortality and the rate of skeletal anomalies among the fetuses. The anomalies are restricted to the anterior part of the axial skeleton and consist essentially in the fusion of 2 or more cervical vertebrae. In addition, lead diminishes the blood calcium levels in the pregnant females. In the calcium deficient group, all these effects of lead are considerably increased and fetuses suffer a loss of weight and delayed ossification. In the animals given such a diet but non lead-injected, the fetal weight is already diminished. However, the ossification and the rate of skeletal anomalies are not affected, and the blood calcium levels of the mothers are similar to those of the control females given a normal diet.


Induction of haemorrhagic necrosis in a murine tumour by products from the culture fluid of Azotobacter.

Macromolecules in the supernatant from cultures of the nitrogen-fixing bacterium Azotobacter chroococcum when given systemically induce permanent regression of established grafts of a syngeneic murine lymphoma. The effect mimics that produced by endotoxin in that haemorrhagic necrosis is induced in the tumour, but the ratio of the toxic dose to the dose needed to cause tumour regression is greater than for this culture supernatant than for endotoxin.


Suppression of DNA synthesis in mitogen stimulated leukocytes with  partial trypsin digests of Staphylococcus aureus protein A.

Staphylococcus aureus protein A (SPA) acts as a mitogen stimulating DNA synthesis and is not antagonistic to the mitogenic activity of other mitogens. The mitogenicity of SPA is due to the protein's multivalent Fc antibody receptors. Partial trypsin digestion of SPA yields monovalent Fc antibody receptors which are not mitogenic. Using a thymidine incorporation assay, results indicated that tryptic digests of SPA added to mouse splenic leukocytes 12 hours after stimulation with either concanavalin A, lipopolysaccaride or SPA suppressed the DNA synthesis normally elicited by these mitogens. SPA was digested with 1% trypsin at pH 8 for varying periods of time. As the digestion time increased from 0 to 60 minutes the mitogenicity of the SPA-trypsin digests decreased indicating cleavage from multivalent to monovalent SPA Fc antibody receptors. The decrease in mitogenicity of the SPA  digests was directly associated with increased ability to inhibit DNA synthesis in mitogen stimulated leukocytes. A proposed mechanism for the inhibition of DNA synthesis suggests that SPA-monovalent Fc antibody receptors mimic cellular Fc receptor function thereby influencing cellular gene expression.


Transplantability and metastatic potential of chemically induced rat brain tumours.

From clinical observations it is known that brain tumours in principle do not metastasize. An explanation for this phenomenon is not available. The few described cases of distant metastases from primary brain tumours all occurred after surgery of the central nervous system. Furthermore, the brain does not contain a lymphatic system. The major question in this matter is whether the inability of CNS tumours to metastasize is based on a specific tumour bound property or on specific local factors. Since an experimental model for this situation was not available we induced brain tumours in rats. About 130 WAG/Rij and Sprague Dawley rats (males and females) were treated with the neurocarcinogen ethylnitroso-urea (ENU) within 24 hours after birth. Tumours appeared at the age of 6 to 29 months. All tumours were removed after killing the host and transplanted subcutaneously into syngeneic rats. Histologically the tumours were mostly oligodendrogliomas, schwannomas and several mixed glial tumours. Metastases from these primary tumours were not observed. The transplanted tumours showed distant metastases in 52% of the cases. Metastases occurred mainly in lungs, liver and lymph nodes. From these observations it is concluded that the absence of metastases from primary brain tumours is probably not related to a specific property of brain tumours. Further research is emphasized on specific local factors.


Effect of thyrocalcitonin on renal reabsorption of bicarbonate.

Porcine thyrocalcitonin (TCT) given intramuscularly at a dose of 2-3 MRC units/kg body weight induces a renal bicarbonate wasting probably at the level of the proximal tubule. This effect is found in normal children as well as in patients with renal tubular acidosis and with vitamin D deficiency. The mechanism by which TCT acts on the acidification mechanism is probably independent of parathyroid hormone (PTH) because the effect could also be obtained in hypoparathyroid patients.


Thymidine labeling index and estrogen receptor level in 64 human breast cancers.

Concurrent assays of labeling index (LI) and estrogen receptors (ER) have been performed in 64 human breast cancers. The dipping method of autoradiography has been used for measurements of LI. The LI values are given as a number per hundred of cell nuclei labeled by tritiated thymidine. ER levels were determined by dextran-coated charcoal method followed by Scatchard analysis, and expressed in femtomoles per milligram of cytosol protein. Tumours with ER greater than or equal to 28 fmol/mg were demonstrating a low LI (less than 2.3%) and come all from patients of age over 50 years. LI and ER values have been compared with clinical data. The best clinical outcome have been found among patients with low LI values (less than 2.3%). Patients with borderline ER values (3 fmol/mg less than ER less than 28 fmol/mg) showed a satisfactory clinical outcome in 88% of cases when LI less than 2.3% and 57% of cases when LI greater than or equal to 2.3%.


Effect of concanavalin A on lysosomal enzyme content of Hela cells.

Precoating Hela cells with concanavalin A (Con A) induces a decrease of lysosomal enzyme intracellular content, and correlatively increases the amount of these enzymes in the culture medium. Possible mechanisms of the lectin effect are discussed in relation to the hypothesis of the secretion-endocytosis cycle of lysosomal enzymes.


Studies of macromolecular heterogeneous associations involving cross-linking: a re-examination of the ovalbumin-lysozyme system.

A system is considered in which a multivalent acceptor interacts with a bivalent ligand in solution to form an array of complexes via multiple binding and cross-linking reactions. With the use of reacted site probability functions expressions are derived in terms of a site binding constant which are of potential use in the interpretation of sedimentation equilibrium and binding results obtained with such systems. Their potential use is explored in relation to results obtained on the interacting ovalbumin-lysozyme system at pH 6.80, ionic strength 0.02. A comparison is made of this interpretation with that based on an interaction pattern involving only multiple binding of ligand in the absence of cross-linking effects. While both interpretations quantitatively describe certain results, it is shown, by invoking further experimental observations on apparent weight-average molecular weight and precipitation behavior, that the more favored interpretation is that involving the operation of a spectrum of forces leading to a large array of ovalbumin-lysozyme complexes, including those of the cross-linked type. It is stressed that the particular ovalbumin-lysozyme system is but one example of interaction between oppositely charged macromolecules and therefore that the derived equations may find wider application to such systems and those known to involve more specific cross-linking interactions.


Identification of carpronium chloride and its metabolite in human urine by field desorption mass spectrometry using deuterium labelling.

The identification of carpronium chloride and a metabolite in human urine has been performed by means of field desorption mass spectrometry using deuterium labelling. Essential points in this study are the simultaneous administration of a deuterium labelled drug ([2H9]carpronium chloride), a purification procedure by ion pair extraction with an iodine reagent, and the use of paper electrophoresis to examine the degree of clean-up. The field desorption mass spectra of the purified extracts obtained from sample urine gave a characteristic pattern resulting from the carpronium cation (m/z 160, m/z 169) and a metabolite of the N-(3-carbohydroxypropyl)trimethyl ammonium cation (m/z 146, m/z 155).


Steroids and hypertension. 1--Gas-liquid chromatography and gas chromatography mass spectrometry of 3,15- and 3, 16-dihydroxy-17-oxosteroids.

In order to analyse and quantitate the urinary 16-oxysteroids known or thought to be associated with hypertension, we have established for six 16-oxy-C19 reference steroids the following parameters: elution volume on lipophilic gel columns, gas chromatographic retention data expressed as methylene unit values of trimethylsilyl ether and O-methoxime trimethylsilyl ether derivatives on OV-1 and OV-17 packed columns and on SE-30 capillary column, and mass spectra of these compounds. These reference steroids were: 3 alpha, 16 alpha-dihydroxy-5 alpha-androstan-17-one, 3 alpha, 16 alpha-dihydroxy-5 beta-androstan-17-one, 3 beta, 16 alpha-dihydroxy-5 alpha-androstan-17-one, 3 beta, 16 alpha-dihydroxy-5-androsten-17-one, 3 beta, 16 beta-dihydroxy-5-androsten-17-one, 3 beta, 17 beta-dihydroxy-5-androsten-16-one and 3 alpha, 15 alpha-dihydroxy-5 beta-androstan-17-one. The proposed method was shown to be applicable to the specific analysis of 16-oxy-C19-steroids in biological samples since it achieved the selective isolation of these compounds from other steroids and their quantitative elution in a single fraction. The analysis of the urinary steroids of two patients with arterial hypertension demonstrated an elevated rate of 3 beta, 16 alpha-dihydroxy-5-androsten-17-one.


Identification and quantitative determination of o- and m-hydroxymandelic acid in human urine.

o-Hydroxymandelic acid and m-hydroxymandelic acid have been identified in human urine by gas chromatography mass spectrometry selected ion monitoring. After solvent extraction the urinary acids were converted to their O-trifluoroacetoxy methyl ester derivatives which were identified by comparison of the retention times and relative intensities of the characteristic m/z 374 and m/z 315 ions with those from authentic samples. 4,6-[2H3]-o-hydroxymandelic acid and 2,4,6-[2H3]-m-hydroxymandelic acid were synthesized for use as internal standards in the quantitative estimation of the isomeric hydroxymandelic acids excreted in urine. In ten normal adults the following results were obtained: o-hydroxymandelic acid 4--16 ng mg-1 creatinine and m-hydroxymandelic acid 11--71 ng mg-1 creatinine. Acid hydrolysis of the urine or ingestion of a diet of known composition did not affect these results, indicating that these compounds are excreted as the free acids and probably arise by metabolism of the corresponding phenylethanolamine(s).


Chemical ionization mass spectrometry of trimethylsilylated carbohydrates and organic acids retained in uremic serum.

After appropriate sample pretreatment and derivatization, uremic serum was investigated by combined high resolution gas chromatography and mass spectrometry, using both electron impact and chemical ionization methods. Electron impact and chemical ionization spectra of a number of identified (trimethylsilylated) carbohydrates and organic acids are compared. The utilization of chemical ionization mass spectrometry, with isobutane as the reagent gas, is discussed in detail. The influence of the reagent gas pressure on the total ion current and on the spectral appearance was studied. The identification of compounds, based on electron impact mass spectral data, was confirmed and often aided appreciably by using this technique. The chemical ionization spectra of trimethylsilyated alditols and aldonic acids, as well as of other organic acids showed protonated molecular ions, whereas aldoses did not. Differences with electron impact spectra are found mainly in the high mass region. The loss of one or more trimethylsilanol groups becomes the predominating fragmentation route at higher reagent gas pressures.


The importance of an elevated skin temperature in transcutaneous oxygen tension measurement.

The oxygen tension as measured by a heated transcutaneous polarographic electrode has been shown by a number of workers to correlate closely with the arterial blood oxygen tension as measured by accepted invasive methods. The present paper presents a study of the dependence of the measured skin oxygen tension values on the temperature of the sensor. A graph is included that emphasizes that the skin sensor temperature should be greater than 43 degrees C if the device is to be operated in the platequ region. The effect of increasing the skin temperature on both the local blood flow and the associated skin permeability changes has been studied. Skin blood flow studies were performed by means of a radioisotope washout technique using krypton-85 and xenon-133 together with a Huch transcutaneous oxygen electrode. The electrode control system incorporates a heating power monitor that Huch and co-workers suggest can indicate the relative blood flow in the tissues beneath the electrode. Two techniques for localized heating of the skin were employed during the washout experiments. First, a copper tube heating ring supplied from a water bath surrounded the area of measurement. Second, a focused source of light applied heat directly to the area of interest, which was 12 mm in diameter. Information obtained in the course of using a quadrupole mass spectrometer with a heated skin probe has given some information about the permeability of the skin and underlying tissues. In these studies three gases were used. Oxygen and argon are transported by the blood, but carbon dioxide tends to be generated within the tissues, and its skin tension is less affected by the blood content. Between 36 degrees C and 45 degrees C it is possible to relate changes in the oxygen and argon skin tensions to changes in capillary blood flow.


A two temperature, two PO2 method of estimating the determinants of tcPO2.

We have prepared an algorithm describing the relationship of tcPO2 to PaO2 and used this with measured values of both tcPO2 and PaO2 under four conditions (two PO2 levels, two temperatures) to generate four equation sets that yielded unique solutions for the four unknown parameters. In adults under a 44 degree C electrode, capillary temperature averaged about 43 degrees C, O2 consumption about 0.0042 ml/gm/min, blood flow about 0.64 ml/gm/min, and diffusion gradient, D, about 32 mm Hg. We have not attempted to define these values in premature or normal newborn infants, because the method requires about one hour of exposure to 100% O2.


Fetal tcPO2 and FHR variability.

The variability of fetal heart rate, expressed as oscillation amplitude, was correlated to fetal tcPO2 in mothers during normal labor without administering analgesics. There was no correlation between tcPO2 levels and FHR variability. Changes in tcPO2 occurred in a few cases simultaneously with FHR variability, but no consistent pattern could be established.


Correlation between fetal scalp tcPO2 and microblood samples.

In summary we would like to state that the correlation between tcPO2 recorded from human fetal scalp and microblood PO2 is quite good. High tcPO2 values are very reassuring for the clinician and indicate a good oxygenation of the fetus. In the presence of a low tcPO2 reading, other signs of fetal hypoxia like abnormal heart rate patterns or a low microblood PO2 are required to confirm the diagnosis. If a low tcPO2 is the only sign suggestive of fetal hypoxia and all other parameters are normal, we recommend removal of the electrode and reapplication in a different spot.


Failure of conventional monitoring to detect apnea resulting in hypoxemia.

In summary, our findings support and confirm the concerns of many investigators that present methods of cardiorespiratory monitoring are inadequate for the detection of many forms of apnea. Nurses underrecord both the frequency and duration of apneic episodes. Bradycardia is an unreliable index of hypoxemia. Thoracic impedance monitors are unreliable because they detect only a fixed duration of respiratory pause and are sensitive to many artifacts unavoidable in a clinical setting. Finally, ineffective breathing patterns such as disorganized breathing, obstructive apnea, and paradoxical breathing are undetectable by thoracic impedance monitoring. We warn against the reliance on heart rate and thoracic impedance monitoring alone for infants with recurrent apnea.


Continuous comparison of in vitro and in vivo calibrated transcutaneous oxygen tension with arterial oxygen tension in infants.

Transcutaneous PO2 simultaneously recorded by Drager and Radiometer electrodes on the abdominal skin was compared for six-hour periods with aortic PaO2 recorded by a Searle intravascular oxygen electrode. Ten newborn infants with serious respiratory illnesses, six of whom needed mechanical ventilation, were studied. The skin electrodes were heated to 44 degrees C and calibrated first in vitro, and then in vivo against the infant's PaO2. The results showed that 1) after in vivo calibration both skin electrodes gave an accurate estimate of PaO2 for six hours without resiting of the electrodes; 2) the Radiometer electrode gave as satisfactory an estimate of PaO2 after in vitro as after in vivo calibration; 3) the Drager electrode gave a significantly less accurate estimate of PaO2 after in vitro than after in vivo calibration; 4) no evidence suggesting that peripheral vasoconstriction interfered with the accuracy of estimation of PaO2 by the skin electrodes was found.


tcPO2 in pediatric cardiology: application during balloon septostomy, tolazoline administration, and in children with right-to-left shunt.

In pediatric cardiology tcPO2 is useful in monitoring cyanotic children given high-risk therapy such as balloon septostomy or drugs with controversial effects such as tolazoline in persistent fetal circulation. tcPO2 during administration of 100% oxygen enables a rapid, noninvasive differentiation between cyanosis due to intracardiac right-to-left shunt and that due to low cardiac output or pulmonary ventilation or diffusion difficulty. The size of the right-to-left shunt can be roughly estimated from the highest value of tcPO2, this estimation being influenced by anemia, hypothermia, and acidosis, among other factors. A trend of the tcPO2 rise is evident 90 seconds after the beginning of oxygen breathing. If tcPO2 does not rise at least 40 mm Hg over the initial value, a significant right-to-left shunt must be suspected. Interpretation of tcPO2 rise is difficult in dynamic right-to-left shunt, changing with oxygen breathing.


Right-to-left ductal shunt and transcutaneous PO2.

The continuous monitoring of the PO2 with double sensors, pre- and post-ductal, is very interesting during the neonatal period; the magnitude of the right-to-left ductal shunting, related to high pulmonary vascular resistance, is well evaluated by the hyperoxemia test. This noninvasive method allows a reliable observation of the pulmonary vascular response to oxygen, alkaline solutions, or tolazoline infusion.


Transcutaneous PO2 monitoring (tcPO2) in the newborn during apneic spells, convulsions, cardiac catheterizations, and exchange transfusions.

Continuous transcutaneous PO2 recording improves the monitoring of critically ill infants and newborns submitted to invasive procedures such as exchange transfusions or cardiac catheterization. The measurement of tcPO2 in babies with apneic spells or seizures gives a better understanding of these respiratory disorders. It is reasonable to recommend the continuous monitoring of PO2 with a transcutaneous electrode in neonatal intensive care units.


The monitoring of adult intensive care patients by transcutaneous PO2 measurements.

In intensive care medicine the tcPO2 measurement can be used efficiently if the problems discussed are taken into consideration. The advantages of the method follow. 1) The trend of the arterial PO2 can be monitored continuously. 2) A large fall in tcPO2 can be a sign of arterial PO2 decrease or of circulatory insufficiency.


Use of the continuous transcutaneous PO2 measurement in lung function tests.

Continuous measurements of transcutaneous PO2 are of interest in pulmonary function tests by which dynamic processes of pulmonary gas exchange are examined. The aim of the present study is to clarify the question of whether the transition function of transcutaneous PO2 after a step change of inspiratory oxygen concentration could give information about pulmonary gas exchange. In 8 healthy volunteers and 31 patients with chronic obstructive pulmonary disease of various degrees the half-value mixing times of transcutaneous PO2 were correlated with parameters of respiratory mechanics and pulmonary gas exchange. In spite of a damping of rapid changes of arterial PO2 by the transcutaneous measurement, there were significant correlations of the mixing times with physiologic right-to-left shunt; with physiologic dead space, alveolar ventilation, and blood gases; and with the parameters of lung overinflation and airway obstruction. There was a significant correlation with the age of the subjects, too. This had to be attributed mainly to an increase in severity of lung function disturbances with advancing age. Aging of the skin seemed to have a minor influence. According to these results, the transition function of transcutaneous PO2 after a step change of inspiratory oxygen concentration is determined mainly by the alveolar ventilation and the ventilation-perfusion inhomogeneity. This paper describes a short test that only takes some minutes of oxygen breathing and that has nearly the same diagnostic value as the measurement of the half-value mixing time.


Interpretation of the tcPO2 curve in adult patients in an intensive care unit.

From the data collected on 135 adult patients in an intensive care unit the following conclusions of the interpretation of the transcutaneous PO2 curve in this type of patient may be drawn: 1) One to four minutes after the electrode was attached a nadir (= the lowest initial point) was reached and then there was a gradual increase which ended with an initial plateau. This plateau could be predicted from the lowest initial point by adding 30 mm Hg. 2) If the lowest initial point was 35 mm Hg or less, it was expected that the final tcPO2 level would be lower than the actual PaO2. 3) No correlation was seen between the lowest initial point of the tcPO2 curve and the time until the initial plateau was reached. In 75% of the cases the plateau was reached within 20 minutes. 4) A slight positive correlation was found between arterial blood pressure and lowest initial point. The difference between the means of the lowest initial point for a group with blood pressure less than or equal to 90 mm Hg and another group with blood pressure greater than 120 mm Hg was highly significant. 5) Transcutaneous PO2 measurements were more likely to represent the arterial PO2 level if blood pressure was greater than 120 mm Hg than if it was less than or equal to 90 mm Hg. 6) The lower the initial tcPO2 level, the more likely it was that the arterial level was higher than the tcPO2 level (see also point 2). 7) The differences between PaO2 and tcPO2 were independent of the arterial PO2 level. 8) The overall correlation between PaO2 and tcPO2 in this material was 0.91; the slope was 0.90 and the intercept -8 mm Hg. The correlation coefficient within each individual patient was higher. 9) The correlation coefficient between PaO2 and tcPO2 for those patients in whom PaO2 was 60 mm Hg or less was comparatively low. This may be explained by the bad condition of these patients followed by a reduced blood flow. 10) An initial drop in tcPO2 to zero level indicates a reduced peripheral circulation, but this must be suspected when the initial drop is below 35 mm Hg.


A comparative study of continuous intravascular O2 saturation and transcutaneous PO2 measurements in the sheep fetus following the reduction of uterine blood flow.

Under physiologic conditions the tcPO2 reflects the PO2 in the carotid artery. It is therefore a measure of the PO2 in the arteries supplying the brain. The response of the tcPO2 to fetal hypoxemia, as during the recovery period following hypoxemia, is delayed. The delayed response of the tcPO2 during recovery is generated by the release of catecholamines into the fetal circulation. The reading of the tcPO2 corresponds better with the course of the heart rate decelerations and the increased blood pressure than with the course of the PO2 in the fetal aorta. The tcPO2 is, in this context, a reliable measure for the peripheral oxygenation and vasoconstriction. The tcPO2 however should be applied with great care for studying the pathomechanism of fetal heart rate decelerations during labor.


Immune complexes and complement in rheumatoid arthritis.

Immune complexes have been shown to occur frequently during rheumatoid arthritis. They have been found in blood, in the synovium and in other extravascular lesions. The recent development of methods for the quantitation of immune complexes provided new tools to evaluate the possible role of immune complexes in rheumatoid arthritis. Immune complexes which appear in synovial fluid are in higher concentration than in serum and have particular physicochemical properties. They likely result from a local formation in the synovium and seem to be directly involved in the generation of the local inflammation. High levels of circulating immune complexes are usually associated with the development of extra-articular vascular lesions. One of the major biological activity of immune complexes is to activate the complement system. There is indeed evidence of complement activation in circulating blood as well as in synovial fluid in patients with rheumatoid arthritis. The presence and the concentration of complement breakdown products in these fluids correlates with the clinical activity. Therefore, the analysis of immune complexes and of complement components appears useful for diagnosis and follow-up, and for the understanding of the pathogenesis of the disease.


[Pathomorphology of human arthroses].

The pathologic morphology of osteoarthrosis (arthrosis deformans) is illustrated macroscopically, histologically, and electron microscopically TEM and SEM). The morphologic hallmark of the lesions is the association of destructive and inadequate reparative processes in the articular cartilage and reactive bone growth, often leading to grotesque deformity of the joints with characteristic osteophytosis. The fundamental morphologic alterations involving chondrocytes and matrix are similar in all arthroses, irrespective of their topography.


Reconstruction of the auricle in minor grades of microtia by a silicone framework.

A silicone ear frame was used in a series of 31 minor microtia cases for subtotal external ear reconstruction. The authors' original method is described and includes the unfolding of the microtic ear in 3 flaps, circular shape trimming of the frame, and enlargement of the conchal cavity. There have been no failures in this series and no exposure in up to 4 years follow-up. Aesthetic results, on the whole, have been excellent.


Reduction mammaplasty using dermofat pedicles.

A technique for reduction mammaplasty is described which aims to preserve the maximum dermal blood and nerve supply to the nipple-areola complex. The procedure is suitable for the correction of all degrees of breast hypertrophy.


Resurfacing denuded areas of the beard with full thickness scalp grafts.

Reconstruction of a beard-bearing area, such as the upper lip, necessitates hair-bearing grafts no thicker than a full thickness skin graft, in order to allow for normal movement. In the normal scalp, however, most of the follicles reach into the subcutaneous fat. By preoperative epilation it seems possible to induce the catagen phase of the hair cycle during which the follicles migrate into the corium and thus will be transplanted as complete morphological units. The clinical results support this concept.


Spontaneous regression of anterior open bite following treatment of macroglossia.

A case of lymphangioma of the tongue with gross mandibular deformity is presented. Surgical reduction of tongue size resulted in considerable remodeling and the orthodontic implications of this remodelling are discussed.


Incidental discovery of recurrent breast carcinoma in patients seeking breast reconstruction.

On the basis of 6 clinical cases where a breast carcinoma metastasis or another breast tumour were discovered during a routine preoperative medical examination or during reconstructive surgery, the author emphasises the role of plastic surgeons in detecting recurrent disease in patients consulting them for reconstructive surgery after breast amputation. Plastic surgeons who probably perform more breast operations than any other specialist should have a thorough knowledge of all the diseases and tumours which may affect these organs.


The failure of free muscle transplants in the treatment of urinary incontinence.

Seven cases of urinary incontinence treated by a free muscle transplant are reported. The incontinence was completely unaffected in 5. One patient  was able to work without using diapers, while another albeit completely incontinent, could voluntarily stop the flow after the bladder was filled with water.


An enquiry into the Mental Health Enquiry.

A limited study into the accuracy of information for the Mental Health Enquiry was undertaken at two major mental illness hospitals in Leicestershire. The results indicate that certain items of data are of questionable value. The current arrangements for processing the data may also need review.


The prediction of the course of minor psychiatric disorders.

Recent studies have suggested that psychosocial factors play an important part in the prediction of the course of minor psychiatric disorders. Fifty-nine new psychiatric out-patients suffering from minor disorders were assessed, using standardized clinical and social interviews, and 52 were followed up after one year and the clinical assessment repeated. Social and clinical factors were equally important predictors of the number of months ill in the survey year, but social and constitutional variables were superior in the prediction of percentage change in symptoms over the year. The results of correlation, factor and multiple regression analyses suggest that the course of minor psychiatric disorder is best predicted by three sets of variables which are, in order of importance, the patient's material social circumstances, his clinical symptoms and his 'genetic risk' scores.


A comparative study of reactive psychosis and acute psychosis without precipitating stress.

A study was carried out to compare the antecedent factors, phenomenology, treatment required and prognosis in 30 cases of Reactive Psychosis and 30 cases of Acute Psychosis without antecedent stress, matched on age and sex. Both groups were followed up for 7 months. It was found that the two groups differed markedly on several dimensions: The Reactive Psychotics had more hysterical and affective features, more vulnerable personality, higher stress experience in their life prior to illness and relatively better prognosis compared to the Acute Psychotics. The difference still remained when those in either of the groups who at the end of the follow-up turned out to be schizophrenics or suffering from affective disorder were excluded from analysis.


Postpartum mania.

Twenty-one patients with bipolar affective disorder (20 manic episodes, one depressive episode) during the postpartum period were evaluated. They were compared to an unselected group of womicantly more Schneiderian symptoms and fewer recurrences of illness within the three-year periofectively ill relatives than the controls. The practical significance of these findings with rey of the bipolar syndrome.


Reported parental characteristics of agoraphobics and social phobics.

The clinical impression that phobic patients perceive their parents as being uncaring and overprotective was investigated in a controlled study of eighty-one phobic patients. Those assigned to a social phobic group scored both parents as less caring and as overprotected, while those assigned to an agoraphobic group differed from controls only in reporting less maternal care. Intensity of phobic symptoms in the pooled sample was examined in a separate analysis. Higher agoraphobic scores were associated with less maternal care and less maternal overprotection, while higher social phobic scores were associated with greater maternal care and greater maternal overprotection.


School phobic children at work.

Seventy-one school phobic children seen at a child guidance clinic were followed up after they had passed school leaving age. Their record in work and further education was compared with the length of time they were off school, whether or not they returned to school, and the occurrence of further psychiatric treatment after discharge from the clinic. It was found that whether or not the child returned to school and the length of time he was off school had no relationship to future performance at work or in further education. A significantly larger number of children receiving further psychiatric treatment after discharge from the clinic failed to reach their potential in work or further education. Adequate treatment of the underlying conditions appears to be more important than returning the child to school.


Alcohol dependence and phobias: clinical description and relevance.

Of 102 alcoholics admitted to an alcoholism treatment unit one third (33) were clinically rated as having a disabling agoraphobia and/or social phobia, and a further third (37) as having less disabling phobic symptoms of either or both kinds. Questionnaires and self-report symptom scales were used to validate the clinical ratings. Alcohol problem screening tests and consumtpion levels confirmed the severity of alcohol dependence. In a group of 44 phobic alcoholics who reports and physical dependence significantly more frequently than the converse. The prevalence of agoraphobia and social phobia in alcoholics is discussed in relation to prevalence date in affective and normal populations, together with their implications for the aetiology of phobias and alcohol det approaches and prognosis.


Should recurrent calcium oxalate stone formers become vegetarians?

The hypothesis that the incidence of calcium stone disease is related to the consumption of animal protein has been examined. Within the male population, recurrent idiopathic stone formers consumed more animal protein than did normal subjects. Single stone formers had animal protein intakes intermediate between those of normal men and those of recurrent stone formers. A high animal protein intake caused a significant increase in the urinary excretion of calcium, oxalate and uric acid, 3 of the 6 main urinary risk factors for calcium stone formation. The overall relative probability of forming stones, calculated from the combination of the 6 main urinary risk factors, was markedly increased by a high animal protein diet. Conversely, a low animal protein intake, such as taken by vegetarians, was associated with a low excretion of calcium, oxalate and uric acid and a low relative probability of forming stones.


Maximum urinary concentration ability in patients with idiopathic hydronephrosis.

The maximum urinary concentration ability and renal parenchymal function of each kidney were investigated in 34 patients with unilateral hydronephrosis. The ability to concentrate urine was not reduced in 10 hydronephrotic and 24 contralateral kidneys. The concentration ability was moderately reduced in 14 hydronephrotic and 10 non-hydronephrotic kidneys and severely impaired in 10 hydronephrotic kidneys. A reduced concentration ability was found almost entirely in hydronephrosis complicated with upper urinary tract infection or renal calculi. Hydronephrotic kidneys without these complications showed a normal concentration ability in 10 of 11 cases. Parenchymal function was reduced in only 10 hydronephrotic kidneys, 7 of which had had upper urinary tract infections and 3 of which had renal stones. It is our opinion that uncomplicated cases of unilateral hydronephrosis should not be operated upon unless necessitated by signs and symptoms. Measurement of the maximum urinary concentration ability might be helpful in setting the correct indication for surgery in borderline cases.


Intravenous inosine for ischaemic renal surgery.

The use of the intravenously administered nucleotide inosine is described for the protection of function in the human kidney undergoing ischaemic renal surgery. The treatment of 7 patients is described. The simplicity of the technique is emphasised.


Technique of clearance (or disconnection) of dilated lymphatics in the renal hilum and lower ureter and bladder in cases of intractable chyluria or haemochyluria.

The indications, investigations and techniques for treating patients with chyluria are presented. Thirty-eight patients were treated by lymphatic disconnection either at renal hilum or at lower ureter and bladder level. Pedal lymphangiography was essential to decide upon the level of operation. Results were satisfactory in the majority when chyluria was intractable.


The underestimated hazards of xanthogranulomatous pyelonephritis.

Twenty-eight cases of xanthogranulomatous pyelonephritis were studied in retrospect and difficulties in diagnosis and treatment are described. Awareness of the entity is the most effective safeguard against over-aggressive resection for this benign condition.


Frusemide intravenous urography in the diagnosis of pelviureteric junction obstruction.

Standard intravenous urography is not a reliable method of demonstrating pelviureteric junction obstruction. Frusemide urography enables 3 groups to be defined on the basis of the percentage increase in the size of the renal pelvis after the diuretic. These groups are clinically significant.


Diuresis renography and the results of pyeloplasty for idiopathic hydronephrosis.

Eighty-six kidneys in 82 patients have been assessed by diuresis renography after pyeloplasty. A non-obstructed diuresis renogram was found in 64% of kidneys. Ninety per cent of patients were clinically improved by surgery and 51% of cases studied had improved urographic features. Thirty kidneys (29 patients) were assessed both before and after pyeloplasty. An improvement in the diuresis renogram was found in 89% of cases postoperatively. Standard renography demonstrated improved drainage in only 33% of kidneys. These findings are discussed in relation to the value of diuresis renography after pyeloplasty.


The early and aggressive repair of iatrogenic ureteric injuries.

Sixty-two iatrogenic injuries of the ureter in 52 patients have been studied over a 15-year period. A plea is made for early recognition in post-operative urinary fistulae and early operative repair. The results of 41 cases treated by the Boari-Ockerblad method are reported without post-operative obstruction or failure.