An interaction between lysozyme and mucus glycoproteins. Implications for density-gradient separations. 1. Some mucus glycoproteins form soluble complexes with lysozyme at neutral pH values. 2. The extent of complex-formation was determined, by an ultracentrifugal difference method, for a range of glycoproteins covering the common blood-group specificities. 3. Interaction was strongest with those glycoproteins of blood-group Lea specificity; these were also richest in sialic acid. 4. Interaction diminished with increase of ionic strength, and was not detectable at I 0.50; however, an asialoglycoprotein was found to retain some activity. The interaction is accordingly primarily, but probably not exclusively, coulombic in origin. 5. The buoyant density of lysozyme in CsCl, CsBr, CsI and Cs2SO4 was determined; the values in the last three salts are anomalously high. This finding accounts for the previously noted difficulty of separating free protein from glycoproteins by single-stage centrifugation in CsBr. 6. Conditions for effective separation of glycoproteins from secretions containing lysozyme by density-gradient centrifugation are reported.


The role of disulphide bonds in human intestinal mucin. Goblet-cell mucin (mucin 1) was isolated and purified from human small-intestinal scrapings. After application of mucin 1 to DEAE-Bio-Gel (A) columns, most of the glycoprotein (76-94% of hexoses) was eluted in the first peak (designated mucin 2). Minor amounts of acidic glycoproteins were eluted with 0.2m- and 0.4m-NaCl in later peaks. Analyses of mucin 1 and mucin 2 revealed mucin 2 to be a monodisperse highly glycosylated glycoprotein containing 6.3% by wt. of protein, N-acetylgalactosamine, N-acetylglucosamine, galactose and fucose. Mucin 1 was similar in composition, but was polydisperse and contained more protein (12.3% by wt.) as well as N-acetylneuraminic acid. Analytical CsCl-gradient ultracentrifugation showed both mucin 1 and mucin 2 to have a major component with an average buoyant density of 1.47000g/ml. Mucin 1 also contained a slightly less-dense minor glycoprotein component. After exhaustive reduction and alkylation mucin 1 retained its major component, but partly dissociated into two lighter glycoprotein components. Mucin 2, in contrast, did not change its density distribution after reduction. Band ultracentrifugation in (2)H(2)O-containing iso-osmotic buffers showed that mucin 1 contained a major fast-sedimenting component (s(o)=37+/-2S), and a minor amount of a slower-sedimenting component. After reduction there was an increased quantity of the latter component, for which an s(o) value of 14.5S was calculated. In contrast, mucin 2 was unaltered by reduction (s(o)=33+/-2S). These findings indicate that the major component of goblet-cell mucin (mucin 2) does not dissociate after S-S-bond reduction, and thus does not apparently rely for its polymeric structure on the association of subunits through covalent disulphide bonds. However, the effects of reduction on mucin 1 suggest that in the native mucin intramolecular disulphide bonds in the minor glycoproteins may stabilize their structure, permitting secondary non-covalent interactions to develop with the major dense mucin (mucin 2) protein.


Studies on the intracellular segregation of polyribosome-associated messenger ribonucleic acid species in the lactating guinea-pig mammary gland. 1. Free and membrane-bound polyribosomes were isolated and the associated mRNA species characterized by cell-free protein synthesis, RNA-complexity analysis and polyribosome run-off in vitro. 2. Of the recovered polyribosomal RNA 85% was associated with membrane-bound polyribosomes and contained 87--93% of the total milk-protein mRNA species as assessed by cell-free protein synthesis or RNA-complexity analysis. 3. RNA-complexity analysis showed that the abundant (milk-protein mRNA assumed) species constituted 55% of the post-nuclear poly(A)-containing RNA population, the remainder consisting of a moderately abundant population (18%) and a low abundance population (27%). Calculations suggest that each population contained up to 2, 48 and 5000 different species respectively. 4. RNA-complexity analysis of the free polyribosomal poly(A)-containing RNA demonstrated that all the species in the post-nuclear fraction were present, though in different proportions, the abundant, moderately abundant and low-abundance groups representing 38, 30 and 32% of this population. 5. RNA-complexity analysis of the membrane-bound polyribosomal poly(A)-containing RNA revealed a more limited population, 72% consisting of the abundant (milk-protein mRNA) species, and 28% a population of up to 900 RNA species. 6. Polyribosome run-off confirmed that milk-protein mRNA was associated with the membrane-bound and free polyribosomes, but represented only a small fraction of the total protein synthesized by the latter. 7. Comparative analysis of milk proteins synthesized in mRNA-directed cell-free systems, or by run-off of free and of membrane-bound polyribosomes, is consistent with the interpretation that in vivo the initiation of protein synthesis occurs on free polyribosomes, followed by the attachment of a limited population to the endoplasmic reticulum. After attachment, but before completion of peptide synthesis, the detachable N-terminal peptide sequence of one of these(pre-alpha-lactalbumin) is removed. 8. The results are discussed in terms of the mechanisms involved in the intracellular segregation of mRNA species in the lactating guinea-pig mammary gland.


The oxidation-reduction potentials of cytochrome o + c4 and cytochrome o purified from Azotobacter vinelandii. Oxidation-reduction titrations of Azotobacter vinelandii cytochrome o + c4 and cytochrome o were performed with simultaneous potential and absorbance measurements under anaerobic conditions. Cytochrome c4 has a midpoint potential (Em, 7.4) of 260mV and purified cytochrome o has an Em, 7.4 of -18mV. Little change in the midpoint potential of cytochrome o was observed when titrated in the pH range 6.2--9.8.


Delineation of fucosyltransferase activities with thiol reagents. The thiol reagent dithiothreitol inhibits the activity of a core GDP-fucose-N-acetylglucosaminide alpha-6-fucosyltransferase in plasma and blood-cell homogenates, while promoting the activity of alpha-2- and alpha-3-fucosyltransferases. The latter enzymes catalyse transfer of fucose on to terminal galactose and subterminal N-acetylglucosamine residues respectively. A thiol-blocking reagent N-ethylmaleimide does not affect the activity of the alpha-6-fucosyltransferase, but inhibits the other two enzymes. These results indicate the presence of a critical disulphide linkage in the alpha-6-fucosyltransferase, and provide a means of delineation of different fucosyltransferases.


The equilibrium assumption is valid for the kinetic treatment of most time-dependent protein-modification reactions. To facilitate mechanistic interpretation of the kinetics of time-dependent inhibition of enzymes and of similar protein modification reactions, it is important to know when the equilibrium assumption may be applied to the model: formula: (see text). The conventional criterion of quasi-equilibrium, k + 2 less than k-1, is not always easy to assess, particularly when k + 2 cannot be separately determined. It is demonstrated that the condition k + 2 less than k-1 is necessarily true, however, when the value of the apparent second-order rate constant for the modification reaction is much smaller than the value of k + 1. Since k + 1 is commonly at least 10(7)M-1.S-1 for substrates, it is probable that the equilibrium assumption may be properly applied to most irreversible inhibitions and modification reactions.


Photoactivated covalent binding of [3H]bilirubin to human serum albumin. A one-step procedure has been developed for the preparation of [3H]bilirubin IX-alpha in good yield from unlabelled bilirubin. Irradiation of an aqueous solution of [3H]bilirubin IX-alpha in the presence of human serum albumin results in the covalent attachment of the bilirubin to the protein. Preliminary degradation studies have been carried out to locate the site of attachment of the bilirubin to the albumin.


Early stimulation of rat liver microsomal protein synthesis after tri-iodothyronine injection in vivo. In an effort to determine the physiological significance of previous studies showing stimulation of microsomal protein synthesis by thyroxine added in vitro, an early effect of tri-iodothyronine injected in vivo was sought. Tri-iodothyronine (25 micrograms/100 g) administered to euthyroid rats stimulated microsomal protein synthesis in vitro within 3--6 h. This effect occurred much earlier than the 26 h lag previously reported after tri-iodothyronine administration to hypothyroid rats. This early effect of tri-iodothyronine on protein synthesis is prevented by alpha-amanitin, suggesting that it is dependent on RNA synthesis. The failure to find a direct effect in vivo of tri-iodothyronine on translation casts doubt on the physiological significance of previous studies that have shown a direct stimulation of translation by thyroxine added in vitro.


Incorporation of N-fluoroacetyl-D-glucosamine into hyaluronate by rabbit tracheal explants in organ culture. 1. Incubation of rabbit tracheal explants with N-[(3)H]acetyl-d-glucosamine and N-acetyl-d-[1-(14)C]glucosamine led to labelling of a number of soluble macromolecular products separable from the medium, after papain digestion, by ion-exchange chromatography. 2. With N-acetyl-d-[1-(14)C]glucosamine in the incubation medium, a neutral glycoprotein, two acidic glycoprotein fractions, hyaluronic acid and a glycosaminoglycan fraction were obtained and all were radioactively labelled. Similar labelling occurred with N-fluoroacetyl-d-[1-(14)C]glucosamine or N-fluoro[(3)H]acetylglucosamine as precursor. 3. Maximal labelling was obtained at 96h after incubation of cultures. N-Fluoroacetyl-glucosamine under these conditions was incorporated into hyaluronate less efficiently than N-acetylglucosamine. 4. With N-fluoroacetyl-d-[1-(14)C]glucosamine as precursor, a hyaluronate component was separated that on enzymic degradation by glycosidases (hyaluronidase, beta-glucuronidase and N-acetyl-beta-hexosaminidase) yielded a (14)C-labelled oligosaccharide fraction together with N-acetyl-d-[1-(14)C]glucosamine and N-fluoroacetyl-d-[1-(14)C]glucosamine, consistent with some exchange of N-acetyl groups having occurred. 5. The results on enzymic degradation of labelled macromolecules by glycosidases suggest that the presence of incorporated N-fluoroacetyl side chains may render the hyaluronate analogue more resistant to hyaluronidase.


Incorporation of exogenous precursors into uridine nucleotides and ribonucleic acid. Nucleotide compartmentation in the renal cortex in vivo. The possibility of compartmentation of UTP in vivo was investigated in the renal cortex of unanaesthetized rats. In addition, liver and spleen were studied in order to compare tissues with different utilization of precursors for pyrimidine nucleotide synthesis. After continuous 2h infusions of [(3)H]uridine or [(3)H]orotate, their incorporation into UTP, UDP-sugars and RNA was quantified. Rates of RNA synthesis were calculated by dividing the incorporation of precursor into RNA by the average specific radioactivity of the UTP pool. Although similar RNA-synthesis rates might have been expected with the two precursors, higher rates were found with uridine than with orotate. The relative incorporation into UDP-sugars of these precursors was also different. Similar results were obtained in the liver. In the spleen, equal amounts of both precursors were incorporated into UTP, but [(3)H]orotate incorporation did not lead to labelling of RNA. To evaluate the heterogeneity of cells with respect to the metabolism of pyrimidines, precursor incorporation was studied in isolated glomeruli and by radioautography. Incorporation into glomeruli was qualitatively similar to but quantitatively different from results in the renal cortex. Although there is obvious tissue heterogeneity, compartmentation of UTP pools is the most credible explanation for the results obtained with the renal cortex and liver. Consequently RNA and UDP-sugars may originate from two different UTP pools. Tissue heterogeneity is the likely explanation for the results obtained in the spleen. Studies of synthesis of pyrimidine and RNA, particularly in relation to growth and regeneration, must take into consideration the precursor used, the apparent existence of UTP compartmentation and the degree of cellular heterogeneity.


Biosynthesis and degradation of prolactin in the rat anterior pituitary gland. Time course of incorporation of label in vitro and evidence for rapid degradation. Biosynthesis of prolactin was studied in anterior pituitary glands from female rats, incubated in vitro. In this system [3H]leucine was incorporated into pituitary proteins, including somatotropin (growth hormone) and prolactin. The rate of uptake of label into prolactin (and to a lesser extent into total protein) slowed considerably during the first 2 h of incubation, although the rate of uptake into somatotropin was constant for 8 h. The most probable explanation for this apparent decrease in the rate of prolactin synthesis is degradation of prolactin in the gland. Degradation of this hormone was also demonstrated by incubating prelabelled pituitaries in unlabelled medium and following the content of labelled prolactin, and by studying the hormonal content of pituitary glands (by radioimmunoassay) before and after incubation. Degradation of prolactin appears to be much more rapid than that of somatotropin, and may represent a physiological mechanism whereby over-accumulation of prolactin is prevented when secretion of the hormone has been rapidly switched off.


A study of the glycerol phosphate acyltransferase and dihydroxyacetone phosphate acyltransferase activities in rat liver mitochondrial and microsomal fractions. Relative distribution in parenchymal and non-parenchymal cells, effects of N-ethylmaleimide, palmitoyl-coenzyme A concentration, starvation, adrenalectomy and anti-insulin serum treatment. 1. GPAT (glycerol phosphate acyltransferase) and DHAPAT (dihydroxyacetone phosphate acyltransferase) activities were measured both in subcellular fractions prepared from fed rat liver and in whole homogenates prepared from freeze-stopped pieces of liver. 2. GPAT activity in mitochondria differed from the microsomal activity in that it was insensitive to N-ethylmaleimide, had a higher affinity towards the palmitoyl-CoA substrate and showed a different response to changes in hormonal and dietary status. 3. Starvation (48 h) significantly decreased mitochondrial GPAT activity. The ratio of mitochondrial to microsomal activities was also significantly decreased. The microsomal activity was unaffected by starvation, except after adrenalectomy, when it was significantly decreased. Mitochondrial GPAT activity was decreased by adrenalectomy in both fed and starved animals. 4. Acute administration of anti-insulin serum significantly decreased mitochondrial GPAT activity after 60 min without affecting the microsomal activity. 5. A new assay is described for DHAPAT. The subcellular distribution of this enzyme differed from that of GPAT. The highest specific activity of DHAPAT was found in a 23 000 gav. pellet obtained by centrifugation of a post-mitochondrial supernatant. This fraction also contained the highest specific activity of the peroxisomal marker uricase. DHAPAT activity in mitochondrial fractions or in the 23 000 gav. pellet was stimulated by N-ethylmaleimide, whereas that in microsomal fractions was slightly inhibited by this reagent. The GPAT and DHAPAT activities in mitochondrial fractions had a considerably higher affinity for the palmitoyl-CoA substrate. 6. Total liver DHAPAT activity was significantly decreased by starvation (48 h), but was unaffected by administration of anti-insulin serum. 7. The specific activities of GPAT and DHAPAT were lower in non-parenchymal cells compared with parenchymal cells, but the GPAT/DHAPAT ratio was 5--6-fold higher in the parenchymal cells.


Detection of peroxisomal fatty acyl-coenzyme A oxidase activity. It has been postulated that the peroxisomal fatty acid-oxidizing system [Lazarow & de Duve (1976) Proc. Natl. Acad. Sci. U.S.A. 73, 2043--2046; Lazarow (1978) J. Biol. Chem. 253, 1522--1528] resembles that of mitochondria, except for the first oxidative reaction. In this step, O2 would be directly reduced to H2O2 by an oxidase. Two specific procedures developed to detect the activity of the characteristic enzyme fatty acyl-CoA oxidase are presented, namely polarographic detection of palmitoyl-CoA-dependent cyanide-insensitive O2 consumption and palmitoyl-CoA-dependent H2O2 generation coupled to the peroxidation of methanol in an antimycin A-insensitive reaction. Fatty acyl-CoA oxidase activity is stimulated by FAD, which supports the flavoprotein nature postulated for this enzyme. Its activity increases 7-fold per g wet wt. of liver in rats treated with nafenopin, a hypolipidaemic drug. Subcellular fractionation of livers from normal and nafenopin-treated animals provides evidence for its peroxisomal localization. The stoicheiometry for palmitoyl-CoA-dependent O2 consumption, H2O2 generation and NAD+ reduction is 1 : 1 : 1. This suggests that fatty acyl-CoA oxidase is the rate-limiting enzyme of the peroxisomal fatty acid-oxidizing system.


Inhibition of gluconeogenesis in isolated rat liver cells by methylenecyclopropylpyruvate (ketohypoglycin). 1. In isolated rat liver cells, hypoglycin is a less effective inhibitor of gluconeogenesis than its transamination product, methylenecyclopropylpyruvate (ketohypoglycin). 2. Methylenecyclopropylpyruvate at 0.3 mM inhibits gluconeogenesis from all substrates tested, except fructose. 3. Methylenecyclopropylpyruvate does not affect 14CO2 release from [1(-14)C]palmitate, but, in the absence of lactate, inhibits ketogenesis and causes a decrease in the [beta-hydroxybutyrate]/[acetoacetate] ratio. These effects are masked when lactate (10 mM) is present. 4. In the presence of lactate and palmitate, 0.3 mM-methylenecyclopropylpyruvate produces a fall in total acid-soluble CoA and a relative increase in short-chain acyl-CoA at the expense of CoA and acetyl-CoA without changing the ATP, ADP and aspartate contents or the [lactate]/[pyruvate] ratio. 5. Many of the effects of methylenecyclopropylpyruvate observed are consistent with inhibition of butyryl-CoA dehydrogenase and of specific CoA-dependent enzymes involved in gluconeogenesis.


Evidence for the operation of the extrahepatic lipoprotein receptor system in vivo in rats. Effect of dietary cholesterol and orotic acid, alone or in combination, on the rate of synthesis of cholesterol and fatty acid in various tissues, measured by using 3H2O. 1. The biosynthesis of cholesterol in vivo was studied at a number of tissue sites in rats by using 3H2O as precursor. Overall, the mass of cholesterol synthesized was in good agreement with the rate of cholesterogenesis, as determined by kinetic analysis of cholesterol specific-radioactivity-time curves after administration of radiolabelled cholesterol. 2. Dietary cholesterol increased the circulating concentration of cholesterol and inhibited endogenous cholesterogenesis, with concomitant increases in the concentration of esterified cholesterol, in all tissues studied. Addition of ororic acid to the cholesterol-supplemented diet tended to reverse each of these changes in extrahepatic tissues. 3. The co-ordinated change in cholesterol biosynthesis and esterification, with no change in total cholesterol content, in extrahepatic tissues, is attributed to control by receptor-mediated lipoprotein uptake. 4. Further reduction of the concentration of the apoprotein B-containing lipoproteins by addition of orotic acid to a diet without supplementary cholesterol did not further enhance cholesterogenesis. We consider that the relatively high rates of extrahepatic cholesterogenesis in normolipidaemic rats are attributable to the low concentration of low-density lipoprotein.


Effects of bivalent cations on prostaglandin biosynthesis and phospholipase A2 activation in rabbit kidney medulla slices. The bivalent cations Ca2+, Mg2+, Co2+, Mn2+, Sr2+ and Ba2+ were compared for their stimulatory or inhibitory effect on prostaglandin formation in rabbit kidney medulla slices. Ca2+, Mn2+ and Sr2+ ions stimulated prostaglandin generation up to 3--5-fold in a time- and dose-dependent manner (Ca2+ greater than Mn2+ congruent to Sr2+). The stimulation by Mn2+ (but not by Sr2+) was also observed in incubations of medulla slices in the presence of Ca2+. Mg2+ and Co2+ ions were without significant effects on either basal or Ca2+-stimulated prostaglandin synthesis. The stimulatory effects of Ca2+, Mn2+ and Sr2+ on medullary generation of prostaglandin E2 were found to correlate with their stimulatory effects on the release of arachidonic acid and linoleic acid from tissue lipids. The release of other fatty acids was unaffected, except for a small increase in oleic acid release. As both arachidonic acid and linoleic acid are predominantly found in the 2-position of the glycerol moiety of phospholipids, the stimulation by these cations of prostaglandin E2 formation appears to be mediated via stimulation of phospholipase A2 activity.


Casein turnover in rabbit mammary explants in organ culture. 1. Explants of mammary gland from mid-pregnant rabbits were cultured in medium 199 containing insulin, prolactin and cortisol, and specific anti-casein immunoglobulin G was used to measure the amount, rate of synthesis and rate of degradation of casein in the explants in the presence of hormones and after removal of hormones from previously stimulated tissue. 2. The amount of casein in particle-free supernatants prepared from mammary explants was measured by ;rocket' immunoelectrophoresis. 3. The rate of incorporation of l-[4,5-(3)H]leucine into casein was measured after isolation of the casein by immunoadsorbent chromatography and polyacrylamide-gel electrophoresis in the presence of urea and sodium dodecyl sulphate. 4. Casein accumulates in mammary explants in the presence of insulin, prolactin and cortisol, but not in the absence of hormones. Removal of hormones after 24h in culture results in a decrease in the rate of accumulation of casein in the explants. 5. Casein-synthetic rate increases in mammary explants in the presence of insulin, prolactin and cortisol, but not in the absence of hormones. Removal of hormones after 24h in culture results in continued casein synthesis at approx. 30% of the rate in the presence of hormones. The synthetic rate does not decrease to values observed in explants cultured throughout in the absence of hormones. 6. Casein is not degraded in mammary explants during a phase of rapid casein accumulation (36-72h) in the presence of hormones. Furthermore casein is not degraded when hormones are removed from the tissue after between 36 and 72h in culture. 7. Casein is glycosylated in mammary explants; the extent of glycosylation parallels the rate of synthesis. The glycosylated protein is rapidly secreted from the tissue. 8. The results are consistent with the notion that after hormonal stimulation mammary explants from mid-pregnant rabbits synthesize, glycosylate and rapidly secrete casein. Removal of hormones decreases the synthetic rate of casein, but does not cause the accumulation of a pool of degradable casein in the lobuloalveolar cells.


Mechanisms of protein degradation in growing and non-growing L-cell cultures. L-cells prelabelled with [14C]leucine and [3H]thymidine were placed in either fresh growth medium (minimal essential medium with 10% serum) or stepdown medium (minimal essential medium) for 3 days. The 14C/3H ratio remained constant in the growing cultures and decreased in the stationary-phase cultures, indicating no protein turnover in growing cultures and a degradative rate of 0.6%/h in the stationary-phase cultures. Media analysis, however, indicated that 14C-labelled proteins were being degraded at approx. 1.2%/h in growing cultures and 1.7%/h in stationary-phase cultures. Additional studies indicated that a subpopulation of L-cells in the monolayer, comprising approx. 20--30% of the total, were lost in the original processing procedure. Experiments in which recoveries approached 100% by fixation of the monolayer in situ indicated that a protein-degrading subpopulation accounted for all the observed proteolysis in the growing cultures. Proteolysis in these cultures was only partially inhibited with NH4Cl, indicating that only a small part of the protein degradation was occurring in an activated lysosomal-autophagic system. NaF produced a more effective inhibition of proteolysis, but we were not able to distinguish whether this effect was on an ATP-requiring basal-turnover mechanism or a direct effect on unregulated activity of proteinases in the cell hyaloplasm. However, NH4Cl inhibited the proteolysis induced when cells were placed in stepdown medium, suggesting that the induced proteolysis was occurring via the autophagic system. We conclude that L-cells exist in at least two states with respect to protein degradation: (a) a subpopulation that is actively replicating and does not degrade cellular proteins, and (b) a second subpopulation of cells, derived from the preceding one, which degraded most of their labelled proteins, are not capable of further replication, and are not sedimented in an iso-osmotic EDTA buffer solution. In addition, proliferating L-cells, when placed in stepdown medium, begin to degrade cell protein through a mechanism involving autophagolysosomes.


The synthesis of myosin, actin and the major protein fractions in rabbit skeletal muscle. New Zealand White rabbits were infused with [3H]tyrosine for periods of 5--6 h and then different methods of extraction were applied for the purification of the main muscle proteins and protein fractions. Myosin (I), prepared from salt extraction of muscle mince, consistently had a higher specific radioactivity than did myosin (II), isolated by dissociation of actomyosin. Actins (IA) and (IB), extracted from acetone-dried powders prepared by different treatments of myosin-extracted muscle mince, gave specific radioactivities approx. 0.6 that of myosin (I) and 0.7 that of myosin(II). Actin (II), isolated by dissociation of actomyosin, had a specific radioactivity similar to that of myosin (II) from the same source, but higher than those of actins (IA) and (IB). The differences between the specific radioactivities of the proteins, in particular actin, purified by the various methods, are attributed to the loss of newly synthesized material of high specific radioactivity during the initial extraction procedures. It is suggested that actin (II) and myosin (II) are representative preparations for the total population of each protein and that, on this basis, myosin and actin have similar rates of synthesis. Total muscle protein, myofibrils, actomyosin and sarcoplasm were all found to have very similar specific radioactivities at the end of a 6 h infusion.


The subcellular distribution of alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase in dog liver. The subcellular distributions of alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase in the particulate fraction of dog liver were examined by centrifugation in a sucrose density gradient. Most of both enzyme activities in the particulate fraction were localized in the mitochondria, but not in the peroxisomes.


N-formyl-L-methionine deformylase activity in human leucocytes and platelets. Extracts of human neutrophils, lymphocytes and platelets enzymically deformylate N-formyl-L-methionine. Enzyme activity is stimulated by Co2+, inhibited by bivalent-cation chelators and unaffected by inhibitors of serine, thiol and carboxyl proteinases. Leucocyte or platelet N-formylmethionine deformylase may be important in modulation of neutrophil responses to chemoattractant formylmethionyl peptides or similar compounds.


Infection in patients with systemic lupus erythematosus. Association with a serum inhibitor of complement-derived chemotactic activity. We have found subnormal amounts of chemotactic activity in zymosan-treated sera from 13 of 29 patients with systemic lupus erythematosus (SLE). As an explanation for this abnormality, the presence of a uniquely specific, heat-stable inhibitor of complement (C5)-derived chemotactic activity has been documented in sera from 11 of these patients. Sera from 2 other patients contained elevated levels of nonspecific, heat-labile chemotactic factor inactivator (CFI) activity. The serum from 1 patient contained the heat-stable inhibitor as well as elevated levels of CFI. Patients with SLE whose sera contained the heat-stable inhibitor had more active disease clinically, but otherwise they were indistinguishable from patients without the inhibitor. When patients with the heat-stable inhibitor improved clinically, this usually was accompanied by a decrease in serum inhibitory activity. Only one episode of bacterial infection was observed among 16 patients with SLE whose sera yielded normal amounts of chemotactic activity after treatment with zymosan. In contrast, 7 of 11 patients with SLE whose sera contained the heat-stable inhibitor suffered serious bacterial infections. The presence of this heat-stable inhibitor in sera from some patients with SLE may contribute, in part, to their increased susceptibility to infection.


Collagen-induced arthritis in rats. Evaluation of early immunologic events. When rats were injected intradermally with an oil emulsion of native type II collagen, they developed an inflammatory polyarthritis. The incidence and severity of arthritis increased as the amount of collagen injected was increased. Rats 4 1/2 weeks old were the most susceptible to the development of arthritis, whereas weanling and older animals were relatively resistant. There was no difference in incidence between males and females. Mononuclear cells from peripheral blood, lymph nodes, and spleen were cultured with type II collagen and responded maximally to a collagen concentration of 25 microgram/ml. The earliest detectable response was in peripheral blood mononuclear cell cultures obtained 6 to 8 days after immunization. The response of lymph node and spleen cells tended to lag behind that of peripheral blood cells at the earlier time intervals. Antibodies were detected in sera by hemagglutination at 8 days postimmunization. Quantitation of IgM and IgG antibodies by radioimmunoassay showed good correlation with hemagglutination titers and increased binding of collagen by both classes of antibody in arthritic as compared to nonarthritic animals. It is clear that the development of both humoral and cellular immunity to type II collagen is associated with the development of arthritis and may be important in the pathogenesis of this disease.


Reaction of superficial bursae in response to specific disease stimuli. Although microscopic studies have shown similarities between bursal and joint membranes, little is known about bursae and their response to disease states. Eighty-six cases of superficial bursitis due to trauma, bacterial infection, or gout were reviewed and compared with cases of joint inflammation due to the same etiologies. In traumatic bursitis the bursal fluid mucin test was more abnormal and the viscosity lower than that of joint fluid in traumatic arthritis. The bursal fluid total leukocyte count of septic bursitis was less than 20,000/mm3 in 8 of 13 cases but in only 1 of 21 synovial fluids from cases of septic arthritis (P = 0.005). In gouty bursitis the mean total leukocyte count of bursal fluid was 2800/mm3, compared with a mean synovial fluid total leukocyte count of 28,700 in gouty arthritis (P less than 0.02). These findings indicate that superficial bursae react less intensely than diarthrodial joints to specific disease stimuli and that a relatively low bursal fluid leukocyte count is often present in cases of septic and gouty bursitis.


Effects of naproxen on connective tissue changes in the adjuvant arthritic rat. The Freund's adjuvant-injected rat shares a number of features with the arthritis patient, viz the presence of a proliferative synovitis, joint swelling, and cartilage and bone erosion. Naproxen, a prostaglandin synthetase inhibitor which is an effective antiinflammatory agent in laboratory animals and humans, was evaluated as an inhibitor of connective tissue destruction in this model by use of radiologic and histopathologic analyses. Sixteen days after rats were injected with Freund's complete adjuvant, marked joint swelling was noted. On day 17, vehicle or naproxen, 7 mg/kg/day, was administered orally. Twenty-eight days later vehicle-treated animals demonstrated the following pathologic changes in their hindpaws; swelling, cartilage loss, large amounts of pannus within the joint spaces, osteoporosis, bone erosions, periosteal new bone formation, heterotopic ossification, and bony ankylosis. Rats treated 28 days with naproxen had significantly milder disease than the vehicle controls. The incidence of severe juxtaarticular bone destruction was 10/10 in the vehicle controls versus 2/10 of the drug-treated group (P less than 0.01). A comparable reduction in cartilage erosion, incidence of pannus, and new bone formation was noted in the drug-treated group. These effects may relate to an inhibition of prostaglandin biosynthesis; prostaglandins have been shown to: 1) stimulate collagenase secretion from macrophages, 2) stimulate bone resorption in vivo and in vitro, and 3) diminish proteoglycan synthesis in cartilage.


Chronic tenosynovial hand infection from Mycobacterium terrae. A 23-year-old healthy man suffered a puncture wound from the fin of a catfish. Chronic tenosynovitis subsequently developed. Synovectomy was performed and cultures yielded Mycobacterium terrae. This organism, usually considered nonpathogenic, has only rarely been associated with human disease. This case further supports the role of this organism as a potential human pathogen and further suggests that M terrae should be considered in aquatic-associated hand tenosynovitis.


The effect of thoracic duct drainage on lymphocyte dynamics and clinical symptoms in patients with rheumatoid arthritis. Thoracic duct drainage (TDD) was performed in 4 patients with severe rheumatoid arthritis. Clinical effects were apparent in all during drainage, but the term of TDD and the cumulative number of lymphocytes drained had no direct relation to the improvement of clinical symptoms. The number of lymphocytes in the peripheral blood increased despite discharge of lymphocytes from the thoracic duct in the very early stage of drainage, suggesting that lymph drainage from thoracic duct accelerates migration of lymphocytes from lymphocyte pools to the blood stream. Biopsy specimens of synovial membranes obtained post-TDD showed marked decrease of mononuclear cell infiltration as compared to the specimens obtained preoperatively. These findings suggest that clinical effectiveness may be due not only to systemic immunosuppression induced by lymphocyte depletion but also to accelerated migration of inflammatory cells from the synovial tissues to the blood stream occurring with dynamic change of lymph flow during TDD.


The relation of antemortem characteristics to cardiovascular findings at necropsy--The Framingham Study. The relation of premorbid cardiovascular risk attributes routinely measures at regular biennial examinations to cardiac necropsy findings using a special autopsy protocol was examined among 127 decedents of the Framingham cohort. Necropsy findings analyzed were: heart weight, left ventricular (LV) muscle thickness, percent luminal insufficiency of the coronary arteries, and percent intimal involvement with atherosclerosis. Clinical data analyzed included weight, height, blood pressure and serum cholesterol measured 1, 5, and 9 years prior to death. Systolic blood pressure, but not diastolic, correlated strongly with heart weight and LV muscle thickness in both sexes, and with atherosclerotic involvement in women, but not men. Serum cholesterol 1, 5, and 9 years antemortem all correlated positively with the degree of luminal insufficiency in men, while in women only cholesterol 9 years before death correlated significantly. In multivariate analysis only systolic pressure in women correlated independently with left ventricular muscle thickness and relative weight was the only independent correlate of heart weight. Only age was an independent significant correlate of the extent of coronary atherosclerosis in women. For men, coronary atherosclerotic involvement was independently correlated with only the serum cholesterol and measures of obesity were the major predictors of heart size. The degree of coronary pathology also correlated positively with heart weight in men and LV muscle thickness in women. In general clinical data measured just prior to death did not correlate as well with pathological findings as did earlier measurements.


Intimal thickening of human femoral arteries with special regard to elastin, Part 1. Diffuse intimal thickening due to growth and age. In infants, human femoral arteries display seam-like internal elastic lamina (IEL) covered with endothelium on the luminal side and with smooth muscle cells (SMC) on the medial side. At birth the growth of IEL is finished, correlated with a loss of microfibrils (MF) at the periphery. With the onset of the postnatal vessel growth the joints of IEL seem to be mechanically widened until they have the appearance of gaps with progressing age. After the age of 40 years there are often rod-like crystallites in the IEL, probably composed of cholesterol esters. A small first consecutive lamina (CL) can be seen already in childhood; it enlarges until the 3rd decade of life and is interpreted as a substitute to the "fragmented" IEL. After the 5th decade of life the first CL is arranged within the intima at a certain distance from the IEL and consisting of loosely arranged elastic fibrils. In very old arteries (beyond the 8th decade of life) gaps are rarely seen in the first CL. In individuals over the age of 30 years, the space between IEL and the first CL is occupied by smooth muscle cells (SMC) which are tightly packed. Additional CLs above the first CL can be found in elderly individuals, there CL obviously contribute to the intimal thickening. The ultrastructure of the elastic elements of the vessel wall and their possible function are discussed.


Production of coronary arteriosclerosis with sex hormones and human chorionic gonadotropin (HCG) in juvenile steelhead and rainbow trout, Salmon gairdneri. Coronary arteries were examined for significant changes in myointimal hyperplasia in one-year old juvenile steelhead or rainbow trout treated with estradiol, testosterone or human chorionic gonadotropin (HCG). In one experiment juvenile steelhead trout were injected weekly with 2 microgram/g HCG, 5 microgram/g estradiol cypionate or 5 microgram/g testosterone cypionate for 50 days. Control fish were those killed prior to treatment or injected with fish saline or cottonseed oil. In two experiments juvenile nonanadromous rainbow trout were treated with estrogen and testosterone with the same weekly doses as juvenile steelhead for 42 and 30 days, respectively. In the first experiment juvenile rainbow were injected weekly with 2 microgram/g HCG. Juvenile rainbow treated with estradiol, testosterone and HCG and juvenile steelhead treated with testosterone had larger coronary lesions than untreated or control fish. Coronary hyperplastic nodules resembled morphologically those observed previously in sexually mature steelhead and rainbow trout. These data support the hypothesis that reproductive hormones are causal factors in spontaneously occurring arteriosclerosis in spawning Salmo gairdneri.


Influence of cholesterol/fat feeding on cholesterol esterification and morphological structures in intestinal mucosa from guinea pigs. Guinea pigs were fed a semisynthetic diet containing 10% (by weight) cottonseed oil and 1% cholesterol. In response to cholesterol/fat feeding there was an increase in both the unesterified cholesterol (UC) and cholesteryl ester (CE) of the intestinal mucosal cell. Along with the increased cholesterol levels there was a 4-fold increase in the microsomal acylCoA:cholesterol acyltransferase (ACAT) activity after only two days of cholesterol/fat feeding. After 6 days on the experimental diet the ACAT activity was up to 8-fold the activity of the control, and then remained at this level for up to 20 days. The increased ACAT activity was probably not due to increased substrate concentration alone, since the fractional esterification of cholesterol also increased when the cholesterol/fat containing diet was given. There was also an increase in the triglyceride content of the intestinal mucosal cells from guinea pigs on the experimental diet. The mucosal cells of the cholesterol/fat fed animals accumulated varying amounts of lipid droplets, which were without an enveloping membrane, suggesting that the uptake of lipids from the intestinal lumen was higher than the capacity to synthesize and/or secrete lipoproteins. Simultaneously the size and amount of secondary lysosomes increased. A considerable increase in lipid droplets, lipolysosomes, and residual bodies was observed in the lamina propria macrophages while no crystalline clefts were seen.


Chronic renal failure and atherogenesis--Serum factors stimulate the proliferation of human arterial smooth muscle cells. To assess the role of serum factors in the genesis of accelerated vascular disease in chronic renal failure, human arterial smooth muscle cells (SMC) and dermal fibroblasts were grown in culture and the effects of serum from chronic dialysis patients on cell proliferation was studied. Exposure to serum from these renal failure patients was associated with significantly greater growth of both SMC and fibroblasts than that observed with control serum. A portion of this mitogenic effect appears to be related to the presence of a factor(s) which is heat stable, dialysable, and is contained in the lipoprotein deficient fraction of plasma of density greater than 1.25 g/dl. These findings suggest that circulating substances which stimulate the proliferation of SMC may contribute to accelerated cardiovascular disease in patients undergoing chronic dialysis treatment.


Turnover of apoE in normal and hypercholesterolemic rats. The turnover of apoE in the total lipoprotein fraction (p less than 1.21 g/ml) of normolipemic and hypercholesterolemic rats was compared. The specific activity of 125I-labelled apoE in the lipoproteins was determined after isolation of the apoprotein by immunoaffinity chromatography. The serum apoE decay curves showed rapid first and slower second phases. The first phase of the curve of hypercholesterolemic animals suggests some sequestration of the apoprotein. The half-lives of apoE in the second phase were approximately 7 and 23 h in the normal and hypercholesterolemic sera, respectively. Elevated apoE levels and moderate hypercholesterolemia persisted one week after restoration of the normal diet, indicating that the increased apoprotein level seen in hypercholesterolemic rats was not solely to VLDL or chylomicron remnants. However, due to the elevated apoE levels in the hypercholesterolemic rats, the total replacement rates of the apoprotein appeared to be greater than that in normolipemic animals, consistent with the concept that in the steady state there is an increase in apoE secretion in hypercholesterolemic animals rather than a decrease in catabolism.


Spontaneous arteriosclerosis in old, male, virgin Sprague--Dawley rats. Virgin, male, Sprague--Dawley rats were observed from 3 to 34 months of age. Systolic blood pressure rose progressively with age, reaching abnormally elevated levels in some, becoming abnormally low in others. The old virgin males gained and then lost weight progressively with age, their pituitary and adrenal glands were heavier than young rats, and their thymi were greatly involuted. The old virgin rats manifested fatty infiltration of the liver and islet beta cell degranulation concomitant with hyperlipidemia, hyperglycemia, and elevated circulating corticosterone levels. Spontaneous arteriosclerosis, identical to that found in younger breeder rats was found in these old virgin males. Therefore, the vascular and other degenerative changes found in young breeder rats in considered to be an example of accelerated arterial disease.


Distribution of intimal smooth muscle cell masses and their relationship to early atherosclerosis in the abdominal aortas of young swine. In the abdominal aortas of young mash-fed swine, intimal cell masses (pads, cushions) are located predominantly away from blood vessel orifices. They are found scattered throughout the aorta but nevertheless have a definite pattern of distribution. In the distal one half of the abdominal aorta, they are more frequent in the ventral quandrant than in the dorsal or either lateral quadrant. In the proximal half, intimal cell masses are more frequent in the dorsal quadrant. When experimental atherosclerosis is induced in the abdominal aortas of young swine by either a hypercholesterolemic diet or by aortic ballooning followed by a hypercholesterolemic diet, the distribution of early lesions is similar. The lesions are found predominantly in quadrants where intimal cell masses were found to be most frequent in the control group of swine. The results suggest that most of the lesions, though not necessarily all, arose from pre-existing intimal cell masses beneath the aortic surface.


Swine in atherosclerosis research--Development of an experimental animal model and study of the effect of dietary fats on cholesterol metabolism. The effect of saturated (beef tallow) and polyunsaturated (corn oil) fat diets on plasma cholesterol, bile flow and composition, and excretion of fecal steroids, was studied in 5 female and 6 male miniature swine. The animals were fitted with a modified Baldwin T-tube in the common bile duct for bile sampling and a catheter in the duodenum for reinfusion. Plasma cholesterol levels were increased by beef tallow and decreased by corn oil. Both fats caused an increase in the secretion of bile, biliary lipids and bile acid pool size. The increases were significant when the source of fat was corn oil. The fecal neutral steroid excretion measured in the male swine was increased by both fats. However, only the polyunsaturated fat diet caused a significant increase in the fecal acidic steroid excretion.


Lecithin:cholesterol acyltransferase activity, plasma and lipoprotein lipids and obesity in men and women. Lecithin:cholesterol acyltransferase (LCAT) activity, lipid concentration, lipoprotein lipid concentrations and cholesteryl ester linoleic acid proportion were determined in the plasma of 85 subjects randomly selected from a population during a health screen survey. Mean fractional LCAT rate was significantly higher in men than in women. Molar LCAT rate correlated with low density lipoprotein (LDL) cholesterol concentration in men and with nearly all lipoprotein  lipid concentrations in women. Most of these relationships were dependent on plasma unesterified cholesterol (UC) concentration. Fractional LCAT rate was correlated only with HDL cholesterol concentration in women and this relation was dependent on the influence of obesity. An inverse relationship between plasma cholesteryl ester (PCE) linoleic acid proportion and molar LCAT rate in women was also explained by influences of obesity on the data. Both fractional and molar LCAT rates were positively correlated with obesity (Quetelet's Index and subscapular skinfold thickness) in women but not in men. This study showed the influence of sex on nearly all correlations involving LCAT activity in combined groups of men and women.


Relationship between inhibition of aortic histamine formation, aortic albumin permeability and atherosclerosis. Effects of partial inhibition of aortic histamine formation on aortic albumin uptake and lipid deposition were examined in male, New Zealand white rabbits maintained on Purina Rabbit Chow containing 0.5% cholesterol for a 2-week period. Aortic histamine synthesis was inhibited by partial inhibition of aortic histidine decarboxylase (HD) through administration of alpha-hydrazinohistidine (alpha-HH, MK785, Regis Chemical Co., 25 mg/kg, i.p. at 12-h intervals). Additional rabbits were maintained on either the cholesterol diet or on Purina Rabbit Chow without cholesterol. Results indicate that administration of alpha-HH for the 2-week period produced a 31% reduction (P less than 0.05) in aortic HD activity in those rabbits maintained on the cholesterol diet, and that concurrently there was a 51% reduction in aorta albumin uptake (P less than 0.025) and a 63% reduction in the extent of oil red O staining. By regression analysis a significant correlation coefficient (r = 0.71, P less than 0.005) was obtained between the aortic albumin uptake and the aortic histamine forming capacity (HFC) in rabbits maintained on this cholesterol diet. These findings indicate that the aortic HD system may be an important enzymatic coupler involved in vascular permeability alterations occurring early in the atherogenic sequence.


Time study of coronary myointimal hyperplasia in precocious male steelhead trout, Salmo gairdneri. Time-related observations of myointimal hyperplasia in precocious male steelhead trout maintained in the laboratory suggest that regression of arterial lesions occurs. The most severe myointimal hyperplasia in these trout was observed from June through August during which time the mean lesion rank of the sampled fish reached 2.3 on a ranking scale of 0-5. This time is concomitant with the period of increasing gonadal mass in the strain studied. However, there was no correlation between gonad size and arteriosclerotic lesion severity. Mean lesion severity of fish examined from September through November was approximately one-half that reported for the previous periods. During the September-November period gonadal weights in this strain of fish decreased. Although no statistical correlations could be derived it is suggested that in steelhead trout, changes in myointimal hyperplasia lesion severity are associated with changing reproductive patterns.


Increased hepatic cholesterol synthesis in normal rats by cross-circulation with ileal bypassed partners. Aortic cross-circulation between Holtzman rat littermates was employed to investigate the possible role of a blood-borne factor from the small intestine in the regulation of hepatic cholesterol synthesis. Experimental pairs, consisting of a normal rat and a distal 50% small bowel excluded partner, demonstrated significantly increased combined hepatic cholesterol synthesis when compared to control pairs, consisting of two normal rats, both at 3 and 5 days following parabiosis. This difference was accounted for by increased hepatic cholesterol synthesis in the normal rat in each experimental pair. Neither weight loss nor differences in dietary intake contributed to this effect. Whole blood cholesterol in the common circulation of both experimental and control pairs was lowered; while hepatic cholesterol content was transiently increased, at 3 but not 5 days following parabiosis. Thus, the intestinal bypassed rat stimulates, or releases inhibition of, hepatic cholesterol synthesis in a non-bypassed parabiotic partner. The mechanism for this phenomenon has yet to be defined.


Atherogenesis in the White Carneau pigeon. Further studies of the role of carbon monoxide and dietary cholesterol. Two experiments are described. In the first, 3 pairs of groups of 20 female White Carneau pigeons were fed on diets containing 0.5%, 1% or 2% cholesterol. Birds in one group from each pair were exposed to 150 ppm carbon monoxide (CO) for 6 h on 5 days of each week for 52 weeks, sufficient to raise their carboxyhaemoglobin (COHb) levels to approximately 10%, while those in the other group were sham-exposed under similar conditions. In the second experiment, 3 groups of 40 female pigeons were each fed on a diet containing 1% cholesterol, one group being exposed to CO to give COHb levels of 20%, one to give COHb levels of 10% and one being sham exposed. In addition, 20 birds in the second experiment were fed on the 1% cholesterol diet but were neither exposed to CO nor sham-exposed. Cholesterol enriched diets caused mean plasma cholesterol values in each group to rise sharply within 4 weeks of starting them, but the levels reached were as high with diets containing 0.5% cholesterol as for diets containing 1% or 2% added cholesterol. Exposure to CO increased plasma and aortic cholesterol levels, though this increase was only statistically significant for aortic levels in the second experiment. In both experiments combined exposure to the 1% cholesterol diet and CO resulted in a significant decrease in aortic triglyceride content. The incidence and severity of coronary artery atherosclerosis was associated with increasing dietary cholesterol. It was also associated with exposure to CO in birds given 0.5% or 1%, but not 2%, dietary cholesterol; the increase in birds given 1% was related to the dose of CO. Possible mechanisms are discussed for this effect of CO, which is not found in normally fed birds.


Lipoproteins and plasma cholesterol esterification in normal and diabetic subjects. Serum lipoproteins, separated by preparative ultracentrifugation and the activity of the plasma enzyme lecithin: cholesterol acyltransferase (LCAT) have been measured in insulin-dependent diabetics, non-insulin-dependent diabetics and in age-matched non-diabetic controls. In the insulin-dependent diabetics, mean total serum cholesterol and high density lipoprotein cholesterol (HDL-C) concentrations were significantly higher than in controls. Non-insulin-dependent diabetics had significantly raised total triglycerides and cholesterol, but HDL-C levels were essentially normal. The increased low density lipoprotein cholesterol (LDL-C) in both diabetic groups was statistically significant in men. A methodological study of HDL separation techniques was carried out to facilitate interpretation of these findings. Mean LCAT activity, by a method reflecting combined enzyme and substrate effects was significantly increased in these diabetic groups. The results confirm recent reports of a raised HDL-C in those insulin-dependent diabetics who are prone to coronary heart disease.


Vascular casting. A new method for the study of the arterial endothelium. Vascular casts of the rat aorta were made using modified Batson's No. 17 anatomical corrosion compound. The outline of the silver-stained endothelial cell boundaries were clearly visible on the cast when examined using both scanning electron and light microscopy. The physical properties of the injectate were determined. The exothermic nature of the polymerization of the injectate did not grossly damage the endothelial cells. A method was developed for recovery, without damage, of the arterial tissue surrounding the cast. The technique of vascular casting appears to be a powerful tool for the study of the arterial endothelium which avoids shrinkage artifacts and maintains 3-dimensional geometry.


Low serum estradiol levels in subjects with arterial calcification. The relationship between arterial calcification and serum estradiol levels was studied in 72 postmenopausal female and 59 male subjects. In both sexes, subjects with iliac artery calcification had rather lower serum estradiol levels (8.4 +/- 1.4 pg/ml in the females, and 19.2 +/- 2.5 pg/ml in the males) than controls (16.1 +/- 1.6 pg/ml in the females, and 29.7 +/- 2.4 pg/ml in the males). The bone mineral content of females with iliac artery calcification (0.44 +/- 0.02 g/cm2) was lower than controls (0.52 +/- 0.01 g/cm2); a positive correlation between serum estradiol levels and bone mineral content was found in the females. However, bone mineral content did not significantly differ between males with and without arterial calcification (0.67 +/- 0.03 g/cm2 in the former, and 0.65 +/- 0.02 g/cm2 in the latter). These results indicate that arterial calcification and increased bone resorption are both individual results of estrogen deficiency.


Angiographic demonstration of recanalization of the basilar artery. This report details the clinical course and angiographic findings of a patient with intermittent vertebrobasilar insufficiency. Recanalization of the basilar artery was demonstrated during anticoagulant therapy. A review of the literature failed to disclose a previous report of this occurrence.


[Vertical transmission of hepatitis B (new personal case in a newborn infant)]. A case of acute B hepatitis in a male 3 month-old baby born to an asymptomatic-HBsAg-carrier mother is reported. There is a good evidence to support the hypothesis of a vertical transmission at delivery. In fact the mother failed to exhibit any reactivity for e/anti-e system and resulted DNA-polymerase negative.


[Diffusion of hepatitis B surface antigens in subjects with bladder schistosomiasis]. The problem of the relationship between the surface B antigen and the parasitosis characterized by active penetration of the larvae through the skin has not yet been resolved. Conflicting results have been reported lately on this problem even though it is currently believed that HBsAg is found more often among patients infected by parasites than among healthy subjects. Serum samples from 67 Somalian patients infected by S. haematobium were tested for the presence of the surface B antigen by ELISA method. The HBsAg was found in 19.4 per cent of these patients, while among controls (90 cases) the frequency of HBsAg was of 10 per cent. A comparative analysis of the results obtained with various methods in Somalia shows that the frequency of HBsAg among subjects with urinary schistosomiasis is of 25.9 per cent with indirect haemoagglutination (IHA), of 19.4 per cent with ELISA method and of 14.8 per cent with radioimmunoassay (RIA). These observations seem to indicate that the problem of an increased frequency of HBsAg among patients with urinary schistosomiasis needs further investigation.


[Antibiotic resistance transmissible in "minor" Salmonella strains recently isolated in the Piedmont region]. The presence of R factors in one hundred (100 strains of Salmonella, recently isolated in Piedmont Region, has been investigated. The resistance to 9 antibiotics has been determined for these strains. 83 proved resistant to one or more antibiotics and in particular 57 of these proved to be multiresistant. Only 18 R factors were put in evidence, 13 of these belonging to Salmonella wien. The majority of the R factors found were present in strains of Salmonella isolated from healthy carriers.


[Influence of composition of the culture media on the in vitro determination of the bacterial sensitivity to sulfonamides, trimethoprim and cotrimoxazole]. The difficulties concerning the interpretation of results of susceptibility tests to sulphonamides, trimethoprim, co-trimoxazole have been investigated. The zones of inhibition produced by discs containing these drugs are peculiar since frequently they contain a light bacterial growth and small colonies. Therefore the real sensitivity or resistance of the microorganism is questionable. Susceptibility tests performed on media of different composition, have shown that the phenomenon is chiefly due to the presence in the medium of thymidine and methionine. Both the substances are end-products of the metabolic pathway on which act sulphonamides, trimethoprim, co-trimoxazole. When they are in the medium, they can be available for the bacteria in spite of the inhibition caused by the mentioned drugs.


Control of endotoxinemia in liver disease by lactulose and paromomycin. Through the use of the Limulus test research has been carried out on gram-negative endotoxin in patients with hepatic cirrhosis, chronic hepatitis, acute hepatitis, and in a control group. The positivity of this test in patients with cirrhosis and chronic hepatitis was 93.3% and in cases of acute hepatitis it was 90.9%. The effect of the combined administration of lactulose and paromomycin on endotoxin blood levels has been evaluated in a group of 9 patients with acute hepatitis, 8 with cirrhosis, 1 in hepatic coma, and 1 patient with chronic persistent hepatitis: in 18 of the 19 patients the Limulus test became negative. The results have been discussed in relation to clinical and laboratory data, and to recent data concerning the interaction between intestinal bacterial flora, endotoxin, and liver. Hypotheses have been proposed regarding the hepatocellular c-AMP mediated mechanism of endotoxin action.


[Changes in the bacterial flora of patients under antineoplastic treatment]. The bacterial flora of the skin from different anatomical sites on cancer patients and a control group of medical personnel was examined. This was done to ascertain if antineoplastic therapy was able to change the pattern of microbial flora of patients and to provide a control for possible infectious complications. The results show that in the control group bacterial flora was normal and the antineoplastic treatment did not succeed in changing the bacterial pattern in the skin of patients deeply. Gram negative bacteria were isolated more frequently from the skin of leukemia patients than from either patients with malignant melanoma or other neoplastic diseases.


Treatment of fulminant viral hepatitis with anti-lymphocyte gamma globulins. Ten patients affected by fulminant viral hepatitis (F.V.H.) were treated with anti-lymphocyte globulins (A.L.G.) besides peritoneal dialysis (P.D.) and exchange blood transfusion (E.T.). Two patients awoke transiently; six recovered from F.V.H., but two of them died afterwards due to later complications. The total of complete recovery was 40%; this result is compared with those previously obtained by the Authors in 38 patients treated only with P.D. and E.T. (28.9% complete recovery), and in seven patients treated with P.D., E.T. and human anti-HBsAg gamma globulins (28.5% complete recovery). The treatment with A.L.G. was performed because of the discouraging results until now obtained with the various liver support systems and is based on the accepted view that lymphocytes might be the true effectors of cell necrosis in F.V.H.


[Cytogenic analysis in HBsAg positive subjects, healthy carriers and patients with acute-phase hepatitis]. A highly significant incidence, ranging from 3% to 20% of chromosomal stickiness and agglutination, has been found in 100% of patients with viral hepatitis HBsAg positive and, in lower grade, in carriers of HBsAg. Whereas, such a structural aberration does not occur in normal subjects. No statistical difference has been found as regard chromosomal fragmentation, presence of incisures and numerical anomalies. A possible role of the observed alterations for an impaired cellular immune function observed in healthy HBsAg carriers has been discussed.


[Effects of ionizing radiations on the ultrastructure of bursa of Fabricius]. The alterations induced in the lymphoid and epitelial cells of the Bursa of Fabricius in chicks exposed at different doses of gamma rays mainly occur during the interfase. The extent of these alterations appears reduced in comparison with that observed in the chick thymus treated in the same way.


[Effects of ionizing radiations on the ultrastructure of the thymus gland]. Different doses of gamma rays induce ultrastructural alterations in limphoid and epithelial cells of the chick thymus which exhibit a different temporal evolution after the treatment. The main changes which occur in lymphoid cells treated with 50 rads disappear 48 hours after the irradiation, while with 100 rads the radiation damage is still evident in epithelial cells.


[Multielectronic computerized EMG: technical notes on the recording and parallel off-line elaboration]. A Multielectrodic EMG analysis program is developing. The purpose is to get as short as possible the main EMG parameters (amplitude, duration, frequency) of most motor units, and to reach an estimation of the anatomical extent of single units. According to the muscle extent a variable number of electrodes are inserted crosswise the fibers. EMG signals are simultaneously recorded on a multichannel AMPEX FR1300 and then off-line processed by a 21MX HP minicomputer connected with a 5Mbytes disc drive. Some technical problems had to be solved:channel amplification adjustment to avoid any difference among preamplifiers calibration and filtering, severe hum filtering of main power that is specially strong in nultielectrodic recording systems, the need of sampling at the same Nyquist time the signals of different channels. The computer is instructed to identify the "sinchronous" units i.e. the motor units recorded from more than one channel. These motor units are detected, counted and deleted from all the channels, except the one where they show the maximum amplitude. The percentage of these sinchronous units depends upon the interelectrodic distance and their anatomical area, thus it can support an evaluation of motor unit anatomical spread.


Effect of temperature on the electrical and mechanical activity of lizard ventricular myocardium. Intracellular action potentials and isometric twitches were recorded from lizard ventricles electrically driven at 20 and 4 beats/min and submitted to temperatures changes between 10.5 and 21 degrees C. It was found that cooling induced a depolarization of the diastolic membrane potential ER, which below 15 degrees C exceeded that predictable for a diffusion potential; on the contrary, during the recovery from hypothermia ER underwent a transitory hyperpolarization. Other effects of the low temperature were a decrease of the maximum rate of depolarization, a lengthening of both the action potential duration and the time to peak contraction, an increase of the strength of contraction, in the hearts driven at 20/min it became apparent also an increase of the action potential overshoot. The hypothesis is discussed that the positive inotropic effect of low temperatures may be due not only to a slowing down of the repolarization of the action potential, but also to an increase of the slow inward current intensity.


Effect of multiple arterial haemorrhage on the coronary circulation. In the open-chest anesthetized dog, multiple arterial haemorrhage induces a reduction of the mean coronary flow, an absote or relative increase of the phasic flow during the early ventricular ejection phase and a decrease during the remaining phases of the cardiac cycle. When blood pressure falls sharply during the haemorrhage, coronary vascular resistance increases, whereas it does not change or decreases when blood pressure falls slowly. Moreover, coronary vascular resistance decreases transiently after the stoppage of the haemorrhage.


Chemical analysis of cryoglobulins observed in a case of essential IgG-IgA cryoglobulinaemia. In a case of essential mixed (IgG-IgA) cryoglobulinaemia the carbohydrate and aminoacid composition of cryoglobulins was chemically analyzed. A normal aminoacid and sialic acid composition was observed, whereas the glucide percentage was augmented with respect to the normal range for the two classes of Ig.


[Basal metabolism and dynamic activity of food in obesity]. Metabolic rate of obese patients is reported, recorded to cellular active mass. The metabolic rate is higher in obese patients and the value is diucthy related to individual actual weight. The dinamic-specific action of proteins is normal.


[Eventual presence of 3-4 benzopyrene in the non-saponificable part of sunflower oil]. Within the limit of the research on the chemical composition of the nonsaponds of the edible oils, under execution in our Institute have been affected some cromatografic gas and spectoskopic analysis (U.V.) on a sample of sunflower seeds oil. Concentrating our research essentially on the determination of the presence of 3-4 benzopyrene in the above oil, we have treated about 8,3 gr. of nonsapond with appropriate chemical methodologies, in order to obtain specific samples for the polycyclic aromatic hydrocarbon analysis. The results of the cromatografic gas and spectoskopic of two fractions, obtained through the process of the preparation of the nonsapond sample, compared with those of a standard solution of 3-4 benzopyrene, exclude the presence of the hydrocarbon in the sunflower oil.


[Free amino acids in calf embryos].

[Free amino acids in calf embryos]. In this work have been reported the research results tending to establish the free amino acidis concentration, existing in the calf embryos. The free embryonal amino acidis composition has been determined through an analysis effected with "Amino Acid Analyzer". Such studies, which conern the researches under execution in our Institute on the chemical composition of the calf embryos furnish the biochemical determination of a biological material, used in some Nations like a commercial product.


[Estrogens in calf embryos].

[Estrogens in calf embryos]. A particular technique has been restated for the analysis of extradiol and extrones. Availing ourselves of such a technique, we have determined the presence of the steroid hormones, extradiol and extrones, in the calf embryos. The above studies are framed within the sphere of the researches made in our Institute, having the scope of determining the biological effects and the chemical composition of the calf embryos, which is utilized like a commercial product in some other Nations.


Dextrocardia--value of segmental analysis in its categorisation. Dextrocardia can be defined as a heart in the right chest with the major axis to the right. This definition, however, conveys no information regarding the chamber arrangements and internal anatomy of the heart. Of 40 patients satisfying this definition in the files of the Brompton Hospital, 33 had angiocardiographic data adequate for complete analysis in terms of connections, relations, and morphology of cardiac segments. They form the subject of this report. There were 16 (48%) patients with situs solitus, 11 (33%) with situs inversus, and six (18%) with situs ambiguus. Of the cases of situs ambiguus, four exhibited laevoisomerism and two dextroisomerism. Of the 16 patients with situs solitus, six had two ventricles and 10 had univentricular hearts; two patients had concordant and three discordant ventriculoarterial connections, seven had double outlet ventricle, and four a single outlet heart. Of the 11 patients with situs inversus, nine had two ventricles and two a univentricular heart of right ventricular type; the arterial connection was concordant in two, discordant in two, double outlet in six, and single outlet in one. Of the six patients with situs ambiguus and laevo or dextroisomerism, four had two ventricles, and two univentricular hearts; the arterial connection was concordant in one, double outlet in three, and single outlet in two. Segmental analysis and the use of basic descriptive terms are essential to define the complex anatomy of such hearts.


Left ventricular function in rheumatic mitral stenosis. Clinical echocardiographic study. Echocardiography was used to examine the extent and significance of impairment in left ventricular function in 20 patients with rheumatic mitral stenosis. Indices of left ventricular performance--normalised mean rate of circumferential fibre shortening (Vcf), ejection fraction, normalised posterior wall velocity, and stroke volume were reduced. The impairment in left ventricular function was related to the degree of functional disability (NYHA), right ventricular dilatation, and left atrial enlargement. Vcf was inversely related to both the internal right ventricular diameter (r=-0.767, P less than 0.001) and the degree of left atrial enlargement (r=-0.554; P less than 0.05). The normalised velocity of the interventricular septum and the maximum systolic and diastolic endocardial velocities were also reduced. These results suggest that abnormalities in contractility of left ventricular myocardium are responsible for the impaired myocardial function in patients with mitral stenosis  and that such impairment is clinically significant.


Ventricular response to isometric and isotonic exercise. Echocardiographic assessment. The effects of supine isometric handgrip and graded isotonic bicycle ergometer exercise on left ventricular performance were studied echocardiographically in 20 normal subjects, aged 18 to 36. Measurements of the left ventricular minor axis diameter were taken from recordings performed at rest, during each form of exercise, and during recovery. At the completion of isometric exercise, the pressure-rate product increased significantly. There was no significant change in percentage of fractional shortening (%deltaD), while there was a small but significant fall in peak velocity of circumferential fibre shortening (peak Vcf). Isotonic exercise resulted in a significant increase in %deltaD and peak Vcf. The pressure-rate product also increased and showed a positive correlation with peak Vcf. Isotonic exercise produced a much greater stimulus to left ventricular contractility than isometric exercise and may be a useful means of detecting latent left ventricular dysfunction echocardiographically.


Spectrum of echocardiographic findings in tricuspid valve endocarditis. This report describes a spectrum of M-mode and cross-sectional echocardiographic abnormalities in eight patients with infective endocarditis of the tricuspid valve. The M-mode echocardiogram of the tricuspid valve was abnormal in all but one patient in whom abnormal echoes were seen anterior to the tricuspid valve, in the right ventricular cavity and right ventricular outflow tract. Six patients had shaggy echoes on the tricuspid valve; and one patient showed multilayered echoes on the tricuspid valve which resembled a right atrial myxoma. Irregular diastolic fluttering of the tricuspid valve, indicative of ruptured chordae tendineae, was noted in three patients. The cross-sectional echocardiogram showed abnormal thick shaggy echoes on the tricuspid valve in all five patients on whom the procedure was performed. Contrast echocardiography confirmed the presence of tricuspid regurgitation in four patients. We conclude that echocardiography is useful in the diagnosis of tricuspid valve vegetations, and in detecting complications such as ruptured chordae tendineae and tricuspid regurgitation.


Long-term results of pulmonary autograft for aortic valve replacement. Eighty-five survivors who left hospital after pulmonary autograft replacement for severe aortic regurgitation have been followed critically. Five patients died in the first five years and 80 were followed for six to 11 years. Important aortic regurgitation occurred only early and was always related to technical malpositioning of one autograft cusp. Seven patients with fascial pulmonary valves had problems, requiring removal in four. There was a small (2%) morbidity from the right sided homograft and six were removed five to seven years later for progressive calcification; three of these had been irradiated. Despite a high incidence of trivial diastolic murmurs this valve replacement is still preferred for young patients without dilated aortic roots since the survivors remain well, with excellent, maintained relief of outflow obstruction, without problems from haemolysis and thromboembolism, and without deteriorating autograft function or need for anticoagulants. Histology of five autografts examined up to seven years after operation has shown normal living architecture.


Comparative method for actuarial analysis of cardiac valve replacements. The use of survival analysis in cardiac surgical reports has been widely accepted in order to characterise and compare performances of different valve prostheses. However, the analyses that have been described elsewhere may be improved in several respects. The original method proposed for prosthetic valve evalation examined follow-up related to patients rather than the valves themselves and therefore neglected the possibility of factors other than the valve itself which could affect the fate of cardiac patients. Furthermore, there has not yet been any application of life table methods to compare two different sets of data, or to estimate the average time before an event occurs. The present work defines valve function and malfunction and separates patient survival from valve performance. We define statistical indices, namely the median remaining lifetime and the instantaneous decrement rate. Both indices can be useful; the former measures the average survival time and the latter focuses attention on the intensity of risk changes. Using established statistical theory we calculate the significance level of difference between two independent survival rates. We qualify the presentation of probabilities with a statement of the associated standard errors.


Thallium-201 myocardial imaging in evaluation of asymptomatic individuals with ischaemic ST segment depression on exercise electrocardiogram. Asymptomatic adults with normal physical examination, normal resting electrocardiogram, and normal routine laboratory evaluation who have a positive exercise electrocardiogram and abnormal exercise thallium-201 myocardial image have a very high probability of angiographically significant coronary artery disease. If, on the other hand, the exercise electrocardiogram is positive for "ischaemic" ST segment changes, but the exercise thallium image is normal, the probability for coronary disease is low. The exercise electrocardiogram combined with thallium-201 myocardial image are safe non-invasive methods which can be performed on an out-patient basis.


Effects of amiodarone on sinus node in man. The effects of amiodarone on the sinuatrial node were studied in 24 patients after an intravenous injection of the drug (5 mg/kg). Sinuatrial function was assessed by rapid atrial pacing and premature atrial stimulation. Sinus cycle length did not change significantly, but the corrected sinus node recovery time was prolonged. While there was no significant change in sinuatrial conduction time, prolongation of the non-reset zone in 14 cases, as well as the abolition of the platesu in 2/24 patients, suggested that conduction of the atrial responses to the sinus node might have been depressed.


Arterial counterpulsation in continuing myocardial ischaemia after acute myocardial infarction. Twenty-six patients underwent arterial counterpulsation for refractory heart failure without shock complicating acute myocardial infarction. Patients were divided into a group of 12 with continuing myocardial ischaemia, evidenced by anginal pain associated with abnormal ST segment elevation, and a group of 14 without continuing ischaemia. Clinical features (apart from pain) and prognostic indices were similar in the two groups when counterpulsation was started but short- and long-term results were different. Hospital survival was 92 per cent (11/12) and 43 per cent (6/14), respectively, in the groups with and without ischaemia and four-year survival was 73 per cent and 7 per cent. Counterpulsation is of greatest value in acute infarction when used to relieve myocardial ischaemia.


Carotid sinus hypersensitivity in patients presenting with syncope. In 23 patients (ages 44 to 81) presenting with syncope, vertigo, or transient amnesia, carotid sinus massage produced a significant bradycardia in association with symptoms. The 10 most severely symptomatic patients were studied electrophysiologically, including measurement of intracardiac conduction times and corrected sinus node recovery times, as well as with carotid sinus massage before and after atropine. The only detectable abnormality in five of this group was asystole produced by carotid sinus massage; the other five had, in addition, evidence of either sinuatrial disease or an intracardiac conduction defect. Cardiac pacing in these 10 patients completely abolished their symptoms. In a control group of 52 asymptomatic patients (ages 36 to 87), an abnormal response to carotid sinus massage was uncommon (2%).


Pericardial effusion complicating psittacosis infection. A 13-year-old schoolboy developed a moderately large pericardial effusion during the course of infection with psittacosis. The value of echocardiography in differentiating between pericardial effusion and myocarditis is shown.


Aortic regurgitation in newborn.

Aortic regurgitation in newborn. Clinical signs of aortic regurgitation were noted after birth in an infant who died suddenly at the age of 18 months. Congenital malformation of the aortic root with an aneurysm of the right aortic sinus and disconnection of the corresponding cusp were found at necropsy.


Calcified left atrial myxoma with floppy mitral valve. A heavily calcified left atrial myxoma in a man aged 53 was diagnosed from the plain chest x-ray film and confirmed by echocardiography. There  was also moderate mitral regurgitation caused by a floppy mitral valve. Before a systemic embolus is considered to have arisen from a floppy mitral valve, echocardiography must be performed.


Asymmetric septal hypertrophy and propranolol treatment in a case of Ullrich-Noonan syndrome. A 4-year-old boy with the Ullrich-Noonan syndrome is described. Asymmetric septal hypertrophy was diagnosed by echocardiography and confirmed at cardiac catheterisation. The aortic subvalvar gradient was reduced from 56 mmHg to 10 mmHg with intravenous propranolol. Relatives of patients with the syndrome should  be screened by echocardiography in the hope that the early detection of asymmetric septal hypertrophy and its treatment with propranolol may reduce the likelihood of sudden death.


Effect of an acute increase in cerebrospinal fluid pressure on lung compliance in the rabbit. Cerebrospinal fluid pressure (Pcsf) was increased acutely by the injection of normal saline into the cisterna magna of anaesthetized rabbits to values which interrupted cerebral flow. When increased Pcsf was maintained for a minimum of 3 min, static lung compliance decreased and lung weight (after sacrifice) increased without marked change in arterial blood-gas tensions. The magnitude of the increase in Pcsf did not affect the change in compliance and weight. There was marked unexplained metabolic acidosis. The pulmonary findings may result from a massive shift of blood from the systemic to the pulmonary circulation leading to increased permeability of the interendothelial junctions of the pulmonary capillaries.


Influence of anaesthetic agents on the survival of rats following acute ischaemia of the bowel. We compared the effects of ketamine and halothane anaesthesia on the survival of rats subjected to superior mesenteric artery occlusion. Survival was significantly greater with halothane. Animals anaesthetized with ketamine developed arterial hypocarbia during bowel ischaemic shock, while those receiving halothane did not. Arterial hypoxaemia was not present in either group. Haematocrits increased in both groups following bowel ischaemia. Excess lactate was increased significantly in non-survivors compared with survivors. The survival data contrast with previous results obtained after haemorrhagic hypotension and emphasize the need to consider differing therapeutic approaches in the various shock states.


Acute effect of fentanyl on haemodynamics and myocardial carbohydrate utilization and phosphate release during ischaemia. The effect of fentanyl 25 micrograms/kg body weight i.v. on left ventricular haemodynamics, myocardial carbohydrate utilization and phosphate release during ischaemia in dogs was investigated. A reproducible degree of ischaemia could be obtained by partial occlusion (stenosis) of the interventricular artery, using an inflatable cuff. Inducing stenosis twice made it possible to use the animal as its own control. Arterio-local venous differences of glucose increased during ischaemia and lactate and inorganic phosphate were released from the ischaemic myocardium. Fentanyl administered before the second stenosis reduced heart rate and, to a lesser extent, mean aortic pressure and left ventricular dP/dt max. The release of lactate and inorganic phosphate was diminished during the period of ischaemia. These findings suggest that fentanyl prevents excessive breakdown of energy-rich phosphates and high anaerobic production rate of lactate by decreasing the energy demand of the ischaemic myocardium.


Effect of nitrous oxide on the cardiovascular system and coronary circulation of the dog. A comparison was made in seven dogs anaesthetized with pentobarbitone of cardiovascular measurements during ventilation with 65-70% nitrous oxide in oxygen and ventilation with 65-70% nitrogen in oxygen. The substitution of nitrous oxide for nitrogen was found to be associated, after 15 min, with a significant decrease in cardiac output and significant increases in right atrial and left ventricular end-diastolic pressure and systemic vascular resistance. There was no significant change in mean coronary artery flow, coronary vascular resistance or myocardial oxygen consumption.


Retrospective analysis of anaesthetics received by patients before susceptibility to malignant hyperpyrexia was recognized. We have reviewed the anaesthetic histories of patients later found to be susceptible to malignant hyperpyrexia (MH). Many patients known to have this condition have been exposed to the known inducing agents previously, sometimes on more than on occasion. It is clear that insusceptibility to MH cannot be assumed from previous uneventful exposure to the known triggering agents and that all patients suspected of having inherited MH must be screened by in vivo muscle biopsy studies. No common factor could be found to explain this phenomenon.


Breath-by-breath halothane monitoring during anaesthesia. A study in children. The use of a halothane meter in routine paediatric anaesthesia with controlled ventilation is described. The results demonstrate the accuracy achieved in the control of the alveolar halothane concentration. Measurement of the alveolar halothane concentration revealed responses to surgical stimulation otherwise obscured by neuromuscular blockade, and this may indicate insufficient depth of anaesthesia.


Ketamine and agenesis of the corpus callosum. Ketamine did not provide adequate anaesthesia for pneumoencephalography in a 10-week-old child with agenesis of the corpus collosum. Associated neurological defects are the most likely reason for this failure.


A modification of the MGA 200 mass spectrometer to enable measurement of anaesthetic gas mixtures. The measurement of anaesthetic gas mixtures with a mass spectrometer is complicated by the overlap of fragmentation products of the anaesthetics with the normal respiratory gas. For the MGA 200 quadrupole instrument the problem has been overcome by incorporating simple circuitry which, for an appropriate mass number, sums the concentrations of the unwanted fragments and subtracts them from the total output for the particular channel; this provides an accurate and stable system (within +/- 1% f.s.d.) over at least 24 h.


Comparative haemodynamic effects of tubocurarine and metocurine in the dog. On accout of its histamine releasing and ganglion blocking properties tubocurarine is known to have significant haemodynamic effects. Methylation of the compound produces metocurine and should decrease both histamine release and ganglionic blockade. The haemodynamic effects of these two compounds were compared in 10 mongrel dogs anaesthetized with chloralose and morphine. Haemodynamic measurements were made 2 min before and 2, 5, 10 and 20 min after administration of the drugs. Each animal received three doses of each drug with a 2-h rest period between doses: tubocurarine 0.35 (muscle twitch ED95), 0.7 and 1.4 mg kg-1 and metocurine 0.2 (2 x ED95), 0.4 and 0.8 mg kg-1. All doses of tubocurarine produced an increase in heart rate (212, 197 and 212% of control respectively). The mean arterial pressure decreased significantly with 0.7 mg kg-1 (48% of control; P less than 0.05). Metocurine produced no significant haemodynamic effects except for the largest dose (8 x ED95). The data suggest that the haemodynamic margin of safety with metocurine in the dog is eight times that of tubocurarine.


Oxygen transport during dopamine infusion in dogs. Catecholamines increase not only oxygen delivery to tissues but also oxygen consumption (VO2). The effect of an infusion of dopamine hydrochloride has been studied at two doses, each in six dogs. Dopamine 10 micrograms kg-1 min-1 caused an increase in haemoglobin concentration and altered cardiac output, oxygen availability and total body oxygen consumption such that oxygen availability ratio increased and (CaO2-CVO2) decreased although these changes were not statistically significant. Dopamine 30 micrograms kg-1 min-1 increased (P less than 0.05) heart rate, haemoglobin concentration and CaO2 and significantly reduced stroke volume and VD/VT. Although oxygen availability increased, increases in oxygen consumption were greater and this resulted in a statistically insignificant reduction in oxygen availability ratio and an increase in (CaO2-CVO2). Terminating the dopamine infusion resulted in significant (P less than 0.05) decreases in cardiac output, PVO2, CaO2), oxygen availability and oxygen consumption and an increase in (CaO2-CVO2). It was concluded that maximum oxygen delivery occurs at a lower dose than that required to produce the maximum increase in oxygen consumption.


Metabolic changes during dopamine infusion in dogs. The effects were investigated, in 12 dogs, of the infusion of dopamine 10 or 30 micrograms kg-1 min-1 on circulating glucose, glycerol, lactate and potassium concentrations. Both doses of dopamine produced in initial increase in blood glucose concentration (P less than 0.05) followed by hypoglycaemia (P less than 0.05). Lipolysis was stimulated as shown by an increase in plasma glycerol concentrations with dopamine 10 micrograms kg-1 min-1 (P less than 0.05) and dopamine 3 micrograms kg-1 min-1 (P less than 0.01). Blood lactate concentrations increased slightly in both groups, but this was significant (P less than 0.05) only in the dogs infused with dopamine 10 micrograms kg-1 min-1. Dopamine had no significant effect on the plasma potassium concentration.


Assessment of the anaesthetic and metabolic activities of dioxychlorane, a new halogenated volatile anaesthetic agent. The ability of dioxychlorane to depress cortical activity in rats with implanted electrodes was compared to that reported previously for methoxyflurane, halothane and enflurane. Dioxychlorane was eight times more potent than enflurane, five times more potent than halothane and twice as potent as methoxyflurane. Serum fluoride concentrations after the administration of dioxychlorane and enflurane were not different from controls. In contrast, serum fluoride concentrations after methoxyflurane reached a value of 105 mumol litre-1 and remained increased for at least the next 48 h. Urine fluoride concentrations in the dioxychlorane and enflurane groups were a half and a quarter, respectively, of those recorded in the methoxyflurane group. Polyuria and polydipsia were observed only in the methoxyflurane group. Dilatation of the proximal convoluted tubules was noted in the rats anesthetized with methoxyflurane. These changes were most marked at the 6- and 24-h periods following anaesthesia. Haemorrhage and ulcerative cystitis were noted in the bladders of the rats subjected to methoxyflurane. Cellular swelling in the proximal tubule was observed in the rats sacrificed 24 h after the administration of dioxychlorane. Enflurane produced no pathological changes.


Percutaneous cannulation of the dorsalis pedis artery. A prospective study. Strain-gauge plethysmography was used to determine the systolic arterial pressure in the great toe of 38 patients (aged 23-70 yr) undergoing lung surgery. In eight patients (21%) manual compression of the dorsalis pedis artery reduced the arterial pressure in the great toe to less than 40 mm Hg, and cannulation of the artery was not attempted. In 24 of 30 patients with adequate collateral arterial supply, a Teflon cannula (Venflon 1.20) was inserted percutaneously to the dorsalis pedis artery. Median cannulation time was 160 min. Six patients (25%, 95% confidence limits 10-47%) developed thrombosis of the artery and, in one, unsuccessful cannulation caused thrombosis. In two patients, recanalization of the artery occurred between the 2nd and the 8th day after operation. In four patients, examination 3-5 months after cannulation revealed a persisting decrease in the function of the dorsalis pedis artery. This suggests that the dorsalis pedis artery should not normally be selected for cannulation.


Cardiorespiratory effects of the sitting position in neurosurgery. The cardiorespiratory effects of raising patients from the supine to the sitting position for neurosurgery were investigated. The change caused marked decreases in cardiac index, stroke volume, PaO2 and Qs/Qt, with a significant increase in (PAO2-PaO2) and total peripheral resistance. It is concluded that patients who maintain normotension in the sitting position do not necessarily have an adequate circulatory status.


Premedication in children undergoing day-care surgery. An oral preparation of dichloralphenazone and paracetamol (Paedosed) was used as an alternative premedication to i.m. morphine and atropine in 562 children undergoing day-care surgery. From a series of observations made, both in hospital and at home, it appears that morphine has several disadvantages when compared with this mixture, provided that the latter is supplemented with i.v. analgesia for the more painful procedures.


Continuous measurement of blood gases in vivo by mass spectrometry. Continuous analysis of oxygen and carbon dioxide tension in the blood phase, over periods of 4-5 h, was carried out in the pulmonary artery (93 determinations in six anesthetized dogs) and in the aorta (29 determinations in four anaesthetized dogs). Silastic-covered stainless steel catheters attached to a mass spectrometer were used. The mass spectrometer signals were linearly related to the blood-gas tensions measured by conventional analysis. The mass spectrometer signals were calibrated in vivo by exposing the animals to high and low oxygen and carbon dioxide tension. With such in vivo calibration the slopes of the straight line regressions for mass spectrometer PO2 or PCO2 on PO2 or PCO2 by conventional analysis were comprised between 0.944 and 1.031 while the standard error of the slopes were between 0.019 and 0.031. Deliberate reductions of cardiac output had little effect on the mass spectrometer readings.


The evaluation of liver function after partial hepatectomy in the rat: serum changes. In serial studies of hepatic function in rats after 70% partial hepatectomy, quantitative changes were found in several of the serum components used clinically to assess liver status. The activities of the following enzymes were found to increase: gamma-glutamyl transpeptidase and lactic dehydrogenase were maximal 6 h postoperatively, while glutamic oxaloacetic transaminase and alkaline phosphatase reached peak values at 24 and 48 h respectively. Albumin levels were found to be relatively constant during the study; however, total protein concentration was lowest 6--12 h postoperatively, paralleling a decrease in globulin concentration. Bilirubin levels were elevated to 4x normal within 12 h after surgery. After partial hepatectomy calcium and phosphorus concentrations were significantly decreased at 24 and 12 h respectively. With the exception of alkaline phosphatase, the activities of all serum components measured returned to normal levels by 1 week  after surgery; the alkaline phosphatase concentration continued to be elevated 2 weeks postoperatively.


Effect of plant lectins on murine myeloma cells. The mitogenic response (determined by uptake of [3H]-thymidine) of two different BALB/c myeloma lines, normal BALB/c spleen cells and spleen cells from tumour-bearing mice in primary culture to PHA, Con A, PWM and Robinia pseudoaccacia (RPA) extract was determined by measuring the uptake of [3H]-thymidine over 96 h. The pattern of the response of tumour and tumour-spleen cells to the 4 different lectins was similar, and different from that of normal spleen cells. The unstimulated cultures of tumour and tumour-spleen cells displayed an initial increased uptake of [3H]-thymidine, whereas stimulated cultures displayed a low initial uptake of label. After an initial phase of inhibition, ADJ-PC5 myeloma cells were stimulated by PHA and PWM to a greater extent than were normal spleen cells. On the other hand, normal spleen cells gave a markedly better response to Con A and RPA. The mitogenic response of tumour-spleen cells to all 4 lectins was intermediate between those observed for tumour and normal spleen cells; they responded better to Con A and RPA than did tumour cells but were not as responsive as spleen cells, whereas the converse was true for their response to PWM and PHA. In agreement with other reports, the data suggest that the responsiveness of tumour-spleen cells was due to the presence of tumour cells in this tissue. These results indicate that there is no definite evidence of an impaired lectin-responsiveness in lymphoid cells of mice bearing a myeloma.


The response of rat bronchus-associated lymphoid tissue to local antigenic challenge. Single doses of antigen suspension (alum-precipitated canine serum proteins) were administered intratracheally to SPF rats. After periods of 1, 2 and 3 weeks rats were killed and their lungs examined histologically. After an initial macrophage and perivascular lymphoid reaction, dose-related increases were found in the amount of bronchus-associated lymphoid tissue (BALT) and in the amount of DNA within BALT cells, indicating increased cell division. Immunoglobulin-containing cells were demonstrated within BALT 3 weeks after the exposure to antigen. A prominent and extensive bronchial lympho-epithelium was seen overlying BALT follicles in antigen-treated rats, while in control animals the respiratory epithelium overlying BALT was predominantly normal ciliated epithelium. The significance of these findings is discussed in the light of the possible defensive role of BALT in cases of respiratory disease of man and animals.