Proteinase inhibitors of bovine nasal cartilage. Extracts from bovine nasal cartilage with 1 M-guanidinium chloride were fractionated by ultrafiltration. Gel chromatography of the low-molecular-weight material resolved three distinct fractions with inhibitory activity against (a) collagenases (22000 mol.wt.), (b) thiol proteinases cathepsin B and papain (13000 mol.wt.), and (c) trypsin and other serine proteinases (7000 mol.wt.).


The formation of lipid-linked sugars by cell-free preparations of lactating rabbit mammary gland. 1. A lactating rabbit mammary-gland microsomal system catalysed the incorporation of mannose from GDP-[U-14C]mannose into three endogenous acceptors, (i) polyprenyl phosphate mannose, (ii) lipid-linked oligosaccharide and (iii) protein. 2. Synthesis of polyprenyl phosphate mannose was stimulated by addition of dolichol phosphate to the incubation medium and was reversed by addition of GDP. The product had properties identical with those of authentic dolichol phosphate mannose. 3. The oligosaccharides derived from acid hydrolysis of the lipid-linked oligosaccharide fraction were of six, eight and nine to ten monosaccharide units, the octasaccharide being the major species formed. The oligosaccharide appeared to be attached to the lipid via a pyrophosphate bridge, since strong alkaline hydrolysis liberated an oligosaccharide phosphate. 4. Polyprenyl phosphate mannose served as a mannose donor to lipid-linked oligosaccharides and protein. When added as exogenous substrate it gave rise to a lipid-linked oligosaccharide of about six units. 5. Incorporation of radioactivity in protein was low, but polyacrylamide-gel electrophoresis of the protein fractions indicated that polypeptides of mol.wts. 115000, 75000 and 33000 were labelled.


Conversion of choline methyl groups through trimethylamine into methane in the rumen. 1. Choline methyl groups were rapidly metabolized to trimethylamine by rumen micro-organisms. 2. Trimethylamine was further metabolized to methane, but this system was more easily saturated by an excess of substrate, so that trimethylamine accumulated in the rumen of the fed animal. 3. Although trimethylamine was the only intermediate isolated in the conversion of the methyl groups of choline into methane, methylamine also served as a substrate for methane production. 4. The methyl group of methionine was also converted into methane by rumen fluid, but the methyl groups of carnitine were not.


Calcium-ion transport by intact Ehrlich ascites-tumour cells. Role of respiratory substrates, Pi and temperature. 1. The interaction of intact Ehrlich ascites-tumour cells with Ca2+ at 37 degrees C consists of Ca2+ uptake followed by efflux from the cells. Under optimum conditions, two or three cycles of uptake and efflux are observed in the first 15 min after Ca2+ addition. 2. The respiratory substrates malate, succinate and ascorbate plus p-phenylenediamine support Ca2+ uptake. Ca2+ uptake at 37 degrees C is sensitive to the respiratory inhibitors rotenone and antimycin A when appropriate substrates are present. Ca2+ uptake and retention are inhibited by the uncoupler S-13. 3. Increasing extracellular Pi (12 to 30 mM) stimulates uncoupler-sensitive Ca2+ uptake, which reaches a maximum extent of 15 nmol/mg of protein when supported by succinate respiration. Ca2+ efflux is partially inhibited at 30 mM-Pi. 4. Optimum Ca2+ uptake occurs in the presence of succinate and Pi, suggesting that availability of substrate and Pi are rate-limiting. K. Ca2+ uptake occurs at 4 degrees C and is sensitive to uncouplers and oligomycin. Ca2+ efflux at this temperature is minimal. These data are consistent with a model in which passive diffusion of Ca2+ through the plasma membrane is followed by active uptake by the mitochondria. Ca2+ uptake is supported by substrates entering respiration at all three energy-coupling sites. Ca2+ efflux appears to be an active process with a high temperature coefficient.


Inhibition of lactate glucogneogenesis in rat kidney by dichloroacetate. 1. Sodium dichloroacetate (1mM) inhibited glucose production from L-lactate in kidney-cortex slices from fed, starved or alloxan-diabetic rates. In general gluconeogenesis from other substrates was no inhibited. 2. Sodium dichloracetate inhibited glucose production from L-lactate but no from pyruvate in perfused isolated kidneys from normal or alloxan-diabetic rats. 3. Sodium dichloroacetate is an inhibitor of the pyruvate dehydrogenase kinase reaction and it effected conversion of pyruvate dehydrogenase into its its active (dephosphorylated) form in kidney in vivo. In general, pyruvate dehydrogenase was mainly in the active form in kidneys perfused or incubated with L-lactate and the inhibitory effect of dichloroacetate on glucose production was not dependent on activation of pyruvate dehydrogenase. 4. Balance data from kidney slices showed that dichloroacetate inhibits lactate uptake, glucose and pyruvate production from lactate, but no oxidation of lactate. 5. The mechanism of this effect of dichloroactetate on glucose production from lactate has not been fully defined, but evidence suggests that it may involve a fall in tissue pyruvate concentration and inhibition of pyruvate carboxylation.


Effect of L-alanine infusion on gluconeogenesis and ketogenesis in the rat in vivo. 1. In 48 h-starved 6-week-old rats the 14C incorporation in vivo into blood glucose from a constant-specific-radioactivity pool of circulating [14c]actateconfirmed that lactate is the preferred gluconeogenic substrate. 2. Increasing the blood [alanine] to that occurrring in the fed state increased 14C incorporation into blood glucose 2.3-fold from [14c]alanine and 1.7-fold from [14c]lactate. 3. When the blood [alanine] was increased to that in the fed state, the 14C incorporation into liver glycogen from circulating [14c]alanine or [14c]lactate increased 13.5- and 1.7-fold respectively. 4. The incorporation of 14C into blood acetoacetate and 3-hydroxybutyrate from a constant-specific-radioactivity pool of circulating [14c]oleate was virtually abolished by increasing the blood [alanine] to that existing in the fed state. However, the [acetoacetate] remained unchanged, whereas [3-hydroxybutyrate] decreased, although less rapidly than did its radiochemical concentration. 5. It is concluded that during starvation in 6-week-old rats, the blood [alanine] appears to influence ketogenesis for circulating unesterfied fatty acids and inversely affects gluconeogenesis from either lactate or alanine. A different pattern of gluconeogenesis may exist for alanine and lactate as evidenced by comparative 14C incorporation into liver glycogen and blood glucose.


The structure and function of glycoproteins synthesized during slime-polysaccharide production by membranes of the root-cap cells of maize (Zea mays). The synthesis of the maize root slime polysaccharides was investigated by using [1-3H]-fucose as a marker for slime production. Three fractions were separated by centrifugation in a CsCl density gradient. Two of these were glycoproteins and occurred within the membranes of the cells of the root tip; the third was the slime polysaccharides. Radioactive pulse-chase experiments showed that the glycoproteins were precursors of the slime polysaccharides, and the carbohydrate portion of the glycoproteins had a similar composition to that of the free slime. The linkage between the protein and the carbohydrate of one of the glycoproteins was shown to be a xylose-threonine bond. It is postulated that the slime polysaccharides are synthesized and transported on proteins within the membrane system of the root tip.


Control of glucose metabolism in isolated acini of the lactating mamary gland of the rat. Effects of oleate on glucose utilization and lipogenesis. Oleate (1mM) had only small inhibitory effects on glucose utilization and lipogenesis in acini isolated from rat mammary gland. Esterification of [1-14C]oleate was unaffected by insulin but were decreased by 60% by acetoacetate (2mM). Glycerol (1mM), but not insulin, relieved this inhibition. These experiments provide further support for the role of acetoacetate in regulating substrate utilization by the gland.


Maintenance of glutathione content is isolated hepatocyctes. 1. During the standard procedure for the preparation of rat hepatocytes, about half of the cellular GSH (reduced glutathione) is lost. 2. This loss is prevented by the addition of 0.1 mM-EGTA (but no EDTA) to the perfusion medium. 3. On incubation with and without EGTA, isolated hepatocytes prepared in the presence of EGTA lose GSH. This loss is prevented by near-physiological concentrations of methionine or homocysteine, but not of cysteine. 4. Cysteine, at concentrations above 0.2 mM, causes a loss of GSH probably by non-enzymic formation of a mixed disulphide. 5. Serine together with methionine or homocystein increases GSH above the value in cells from starved rats in vivo. This is taken to suggest that cystathionine may be a cysteine donor in the synthesis of gamma-glutamylcysteine, the precursor of GSH.


Protein degradation rates in regions of the central nervous system in vivo during development. The rate of protein degradation was estimated in several regions of rat brain at various ages by subtracting the rate of accumulation of protein from the rate of synthesis. The rate of degradation in cerebral hemisphere, which was 1.3%/h at 2 days of age, declined steadily with age, approaching the synthesis rate is about 30 days of age (0.8%/h). Degradation rates in the pons medulla, mid-brain and spinal cord were of a similar order to that in the cerebral hemisphere. The cerebellum had an exceptionally high rate of degradation in young rats, 1.9%/h at 2 days of age, which complemented its high rates of synthesis and accumulation. The degradation rate in the young was 2-2.5 times the rate in older rats and was approx. 65% of the synthesis rate during the more active phase of growth. The rapid accumulation of protein in the nervous system during the first week post partum was accompanied by high rates of breakdown, and was the result of a relatively small difference between that high rate of degradation and an even higher synthesis rate.


Evidence for lysosomal enzyme recognition by human fibroblasts via a phosphorylated carbohydrate moiety. Adsorptive endocytosis of five different lysosomal enzymes from various human and non-human sources was susceptible to inhibition by mannose and l-fucose, methyl alpha-d-mannoside, alpha-anomeric p-nitrophenyl glycosides of mannose and l-fucose, mannose 6-phosphate and fructose 1-phosphate. A few exceptions from this general scheme were observed for particular enzymes, particularly for beta-glucuronidase from human urine. The inhibition of alpha-N-acetylglucosaminidase endocytosis by mannose, p-nitrophenyl alpha-d-mannoside and mannose 6-phosphate was shown to be competitive. The loss of endocytosis after alkaline phosphatase treatment of lysosomal enzymes supports the hypothesis that the phosphorylated sugars compete with a phosphorylated carbohydrate on the enzymes for binding to the cell-surface receptors [Kaplan, Achord & Sly (1977) Proc. Natl. Acad. Sci. U.S.A.74, 2026-2030]. Endocytosis of ;low-uptake' forms of alpha-N-acetylglucosaminidase and alpha-mannosidase was likewise susceptible to inhibition by sugar phosphates and by alkaline phosphatase treatment, suggesting that ;low-uptake' forms are either contaminated with ;high-uptake' forms or are internalized via the same route as ;high-uptake' forms. The existence of an alternative route for adsorptive endocytosis of lysosomal enzymes is indicated by the unaffected adsorptive endocytosis of rat liver beta-glucuronidase in the presence of phosphorylated sugars and after treatment with alkaline phosphatase.


Effects of aliphatic diamines on rat liver ornithine decarboxylase activity. Rat liver ornithine decarboxylase activity was decreased by administration of putrescine (1,4-diaminobutane) or other diamines, including 1,3-diaminopropane, 1,5-diaminopentane and 1,6-diaminohexane. This effect was seen in control rats and in rats in which hepatic ornithine decarboxylase activity had been increased by administration of growth hormone (somatotropin) or thioacetamide. Loss of activity was not dependent on the conversion of putrescine into polyamines and was short-lived. Within 6h after intraperitoneal administration of 0.8 mmol/kg body wt., ornithine decarboxylase activity had returned to normal values. This return correlated with the rapid loss of the diamines from the liver, and the decrease in activity could be slightly prolonged by treatment with aminoguanidine, a diamine oxidase inhibitor. A decrease in ornithine decarboxylase activity by these diamines was accompanied by the accumulation in the liver of a nondiffusible inhibitor that decreased the activity of a purified ornithine decarboxylase preparation. The possibility that administration of non-physiological diamines that are not converted into polyamines might be useful for the inhibition of polyamine synthesis is discussed.


Stimulation of precipitation of calcium phosphate by matrix vesicles. The ability of matrix vesicles isolated from the epiphysial growth plate of 6-week-old chicks to facilitate the precipitation of calcium phosphate was studied in vitro. The vesicles lowered the minimum concentration product [ca2+]X[p1] needed to induce crystal formation, thereby showing the vesicles are nucleators of crystallization. After freezing and thawing the vesicles at pH6.0, part but not all of this ability to nucleate disappeared. Freezing and thawing markedly decreased the Ca and Pi content of the vesicles, suggesting that part of the nucleating activity may have been due to mineral already present. After removal of the mineral the residual nucleating activity could be destroyed by extracting the vesicles with lipid solvents or by treatment with enzymes such as phosphoilipase C, neuraminidase or proteinase. Matrix vesicles obtained from chicks treated with 1-hydroxyethane-1, 1-diphosphonate, a compound that inhibits calcification in vivo, showed impaired nucleating activity, both before and after treatment at pH6.0. The vesicle preparation bound some diphosphonate in vitro, probably to the mineral present in the preparation, since no binding could be detected in vesicles preincubated at pH6.0. No difference was found in the nucleating activity of vesicles isolated from rachitic chicks which had or had not received cholacalciferol 48 h before death. These results suggest that matrix vesicles possess intrinsic nucleating activity that may be important in biological calcification.


Prolone metabolism in isolated rat liver cells. The metabolism of proline was studied in liver cells isolated from starved rats. The following observations were made. 1. Consumption of proline could be largely accounted for by production of glucose, urea, glutamate and glutamine. 2. At least 50% of the total consumption of oxygen was used for proline catabolism. 3. Ureogenesis and gluconeogenesis from proline could be stimulated by partial uncoupling of oxidative phosphorylation. 4. Addition of ethanol had little effect on either proline uptake or oxygen consumption, but strongly inhibited the production of both urea and glucose and caused further accumulation of glutamate and lactate. Accumulation of glutamine was not affected by ethanol. 5. The effects of ethanol could be overcome by partial uncoupling of oxidative phosphorylation. 6. The apparent K(m) values of argininosuccinate synthetase (EC 6.3.4.5) for aspartate and citrulline in the intact hepatocyte are higher than those reported for the isolated enzyme. 7. 3-Mercaptopicolinate, an inhibitor of phosphoenolpyruvate carboxykinase (EC 4.1.1.32), greatly enhanced cytosolic aspartate accumulation during proline metabolism, but inhibited urea synthesis. 8. It is concluded that when proline is provided as a source of nitrogen to liver cells, production of ammonia by oxidative deamination of glutamate is inhibited by the highly reduced state of the nicotinamide nucleotides within the mitochondria. 9. Conversion of proline into glucose and urea is a net-energy-yielding process, and the high state of reduction of the nicotinamide nucleotides is presumably maintained by a high phosphorylation potential. Thus when proline is present as sole substrate, the further oxidation of glutamate by glutamate dehydrogenase (EC 1.4.1.3) is limited by the rate of energy expenditure of the cell.


The identification of glycoproteins associated with elastic-tissue microfibrils. Cell cultures derived from foetal bovine ligamentum nuchae accumulate extracellular fibrils morphologically identical with elastic-tissue microfibrils. Two glycoproteins synthesized by the ligament cells are closely related to the matrix microfibrils as assessed by immunological and chemical extraction techniques.


Self-association of proteoglycan subunits from pig laryngeal cartilage. Proteoglycans from pig laryngeal cartilage prepared by dissociative extraction in guanidine hydrochloride were studied in dilute solution by light-scattering and ultracentrifugation. In buffered 150mM-NaCl, pH7.4, the proteoglycan particle weights were about 5x10(6) daltons, but at 100mM-, 200mM- and 300mM-NaCl particle weights of 2.5x10(6)--3.0x10(6) daltons were observed. These results, together with corroborative evidence from sedimentation-velocity experiments, were interpreted in terms of proteoglycans self-associating at physiological ionic strength. The data were examined by using a proteoglycan monomer-dimer model. Proteoglycan preparations that had thiol groups partially carboxymethylated gave particle weights of 3.2x10(6)--3.5x10(6) daltons in 150mM-NaCl, which suggested that carboxymethylation inhibited multimerization and hence that the protein core is implicated in the binding site. Further studies showed that the multimers were stable to 60 degrees C, unlike the hyaluronate-proteoglycan complex.


Cytosol oestrogen receptor of lactating mammary gland. Effect of heparin on the aggregation of the receptor and interaction of the receptor with heparin-Sepharose. 1. An oestrogen receptor is present in low-salt cytosol of the mammary gland of lactating mice as a large aggregate; it is excluded from gel matrix when filtered on a Sephadex G-200 column and sediments at 7S in sucrose gradients. After incubation of cytosol with heparin, the receptor is dissociated. On a Sephadex G-200 column, it is included in the gel matrix and eluted as a protein with mol.wt. 260000 and a Stokes radius of 6.8nm; it sediments at 6S in sucrose gradients. 2. Dissociation of the mammary-gland cytosol oestrogen receptor seems to be the result of interaction of the oestrogen-receptor complex with heparin. This receptor interacts with heparin covalently bound to Sepharose, thereafter sedimenting at 6S. By using this interaction, the cytosol receptor was purified 200-fold compared with the homogenate, with a yield of 70%. 3. The cytosol receptor that was not incubated or was incubated with heparin was much smaller during sucrose-gradient centrifugation than during gel filtration. This discrepancy can be explained by pressure-induced dissociation during high-speed centrifugation. This possibility is supported by the decrease in the sedimentation coefficient of the receptor with increased duration of centrifugation.


Renal inactivation of substance P in the rat. The activity and distribution of substance P-catabolizing enzyme(s) were studied in the rat kidney. Kidney homogenates inactive substance P 5-20 times as fast as do homogenates of intestine, liver, lung, heart or brain. The catabolizing activity was highest in the cortex and decreased progressively down the papilla. Cortex of rat kidney was homogenized and fractions enriched in microsomal membrane, final supernatant, plasma membrane, endoplasmic reticulum, brush border and intact glomeruli were prepared. The identity and homogeneity of the preparations were determined by assaying marker enzymes and by morphological examination. Substance P was catabolized most rapidly by the microsomal and plasma-membrane-enriched fractions, and least rapidly by endoplasmic reticulum or final supernatant fractions. Purified brush border of proximal tubules inactivated substance P more than 10 times as fast as isolated glomeruli. Our experiments show that substance P is catabolized at a rate that is similar to the rates of inactivation of bradykinin and angiotensin II. Further, the distribution of substance P-catabolizing activity in various kidney fractions is similar to the distribution of kininase and angiotensinase activities previously reported.


Maximizing the purification of the activated glucocorticoid receptor by DNA-cellulose chromatography. With heat treatment (20 degrees C for 30 min), the glucocorticoid-receptor complex becomes 'activated' and undergoes an increase in affinity for DNA. A two-stage procedure was used to separate sequentially the rat liver glucocorticoid-receptor complex from proteins with high and low affinity for DNA. DNA-cellulose column chromatography of unheated cytosol resulted in the retention of DNA-binding proteins, but not the unactivated receptor complex. Heat treatment of the column eluate resulted in increased affinity of the receptor complex to DNA, and chromatography on DNA-cellulose then yielded receptor complex free from proteins with low affinity for DNA. Removal of DNA-binding proteins during the first chromatographic step was critically dependent on ionic conditions and the ratio of cytosol chromatographed to DNA-cellulose. A purification of 11000-fold (85% yield) was achieved by this procedure. The partially purified receptor complex was taken up by rat liver nuclei.


Glucuronic acid conjugates of bilirubin-IXalpha in normal bile compared with post-obstructive bile. Transformation of the 1-O-acylglucuronide into 2-, 3-, and 4-O-acylglucuronides. Structures have been determined for bilirubin-IXalpha conjugates in freshly collected bile of normal rats, dogs and man and in post-obstructive bile of man and rats. The originally secreted conjugate has been characterized as azopigment (I), i.e. a 1-O-acyl-beta-d-glucopyranuronic acid glycoside. Conversion of the acetylated methyl ester of azopigment (I) into methyl 2,3,4-tri-O-acetyl-1-bromo-1-deoxy-beta-d-glucopyranuronate (V) indicates the pyranose ring structure for the carbohydrate and a C-1 attachment for the bilirubin-IXalpha acyl group. Alternative procedures for deconjugation of azopigment (I) and its derivatives are also described. In post-obstructive bile, the 1-O-acylglucuronide is converted into 2-, 3- and 4-O-acylglucuronides via sequential intramolecular migrations of the bilirubin acyl group. The following approach was utilized. (1) The tetrapyrrole conjugates were cleaved to dipyrrolic aniline and ethyl anthranilate azopigments, and the azopigments were separated as the acids or methyl esters. (2) The isomeric methyl esters were characterized by mass spectral analysis of the acetates and silyl ethers. (3) The free glycosidic function was demonstrated by 1-oxime and 1-methoxime derivative formation. (4) The position of the dipyrrolic O-acyl group was determined for the methyl esters by protecting the free hydroxyl groups of the glucuronic acid moieties as the acetals formed with ethyl vinyl ether and by further conversion of the carbohydrates into partially methylated alditol acetates. These were analysed by using g.l.c.-mass spectrometry. The relevance of the present results with regard to previous reports on disaccharidic conjugates is discussed. Details of procedures for the formation of chemical derivatives for g.l.c. and mass spectrometry have been deposited as Supplementary Publication SUP 50081 (15 pages) at the British Library Lending Division, Boston Spa, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1978), 169, 5.


DNA polymerases from Chlamydomonas reinhardii. Further characterization, action of inhibitors and associated nuclease activities. The properties of three DNA polymerase species A, B and C, purified from Chlamydomonas reinhardii were compared. DNA polymerases A and B have Km values with respect to deoxyribonucleoside triphosphates of 19 micron and 3 micron respectively. DNA polymerase A is most active with activated DNA, but will also use native DNA and synthetic RNA and DNA templates with DNA primers. DNA polymerase B is also most active with activated DNA, but will use denatured DNA and synthetic DNA templates. It is inactive with RNA templates. DNA polymerase B is completely inactive in the presence of 100 micron-heparin, which has no effect on DNA polymerase A activity. Heparin dissociates DNA polymerase B into subunits that are still catalytically active, but which heparin inhibited. DNA polymerase B possesses deoxyribonuclease activity that is inhibited by 5 micron-heparin, suggesting that the deoxyribonuclease is an integral part of the DNA polymerase moiety. DNA polymerase A is devoid of nuclease activity. DNA polymerase C is similar to DNA polymerase B in all these properties, though it is more active with RNA primers and has greater heat-sensitivity.


Kinetics of tetrahydrobiopterin synthesis by rabbit brain dihydrofolate reductase. Product identification and kinetic data are presented for the conversion of 7,8-dihydrobiopterin into tetrahydrobiopterin by purified rabbit brain dihydrofolate reductase.


A study of the role of the reactive thiol group of rabbit muscle creatine kinase with a chromophoric reporter group. Substrate- and ligand-induced conformational changes were studied in a series of thiol-modified derivatives of rabbit muscle creatine kinase that retained different amounts of enzymic activity. The results indicate that the 'reactive' thiol group of the enzyme is required for the conformational changes associated with formation of a 'transition-state analogue' complex.


The intracellular location of pyruvate carboxylase, citrate synthase and 3-hydroxyacyl-CoA dehydrogenase in lactating rat mammary gland. The intracellular location of pyruvate carboxylase (EC 6.4.1.1), citrate synthase (EC 4.1.3.7) and 3-hydroxyacyl-CoA dehydrogenase (EC 1.1.1.35) in rat mammary gland was investigated by using a fractional-extraction technique. The results indicate a mitochondrial location for all three enzymes.


Crystallization of human oxyhaemoglobin from poly(ethylene glycol) solutions. Crystals of human oxyhaemoglobin were obtained from poly(ethylene glycol) solutions. Spectroscopic and spectrophotometric measurements on the solutions during crystallization and on the dissolved crystals indicate that the method yields stable crystals of oxyhaemoglobin. Preliminary X-ray studies showed that the crystals obtained are isomorphous with those of deoxyhaemoglobin obtained from poly(ethylene glycol) solutions [Ward, Wishner, Lattman & Love (1975) J. Mol. Biol. 98, 161-177].


Conformational variation in superhelical deoxyribonucleic acid. Sedimentation experiments have shown that superhelical DNA undergoes a sharp structural transition at low ionic strength. Light-scattering experiments show that this is due to a change in conformation of the DNA rather than to a change in interactions among DNA molecules. The results show that two possible conformations can occur for superhelical DNA under routine experimental conditions and may explain the discrepancies in the number of early unwinding sites exposed by different techniques.


A simple high-yield procedure for isolation of human urinary kallikreins. Two human urinary kallikreins (fractions A-1 and A-2) were purified to apparently homogeneous forms. The two kallikreins were separated by affinity chromatography using Trasylol (aprotinin) covalently bound to Sepharose. The kallikreins were eluted with a pH gradient (pH 9.5-3.0). Fraction A-1 was eluted between pH 6.2 and 4.2 and fraction A-2 was eluted between pH 4.2 and 3.1. Final purification was obtained by chromatography on Sephacryl SS-200. Antibodies prepared against fraction A-2 were also reactive with fraction A-1; thus it may be possible to measure both by radioimmunoassay using the same antibody preparation.


Characterization of monoferric fragments obtained by tryptic cleavage of bovine transferrin. 1. The electrophoretically fast (F) and slow (S) fragments obtained by tryptic cleavage of bovine iron-saturated transferrin differed in carbohydrate content and peptide 'maps'. 2. A fragment capable of binding one Fe3+ ion per molecule was isolated after brief tryptic digestion of bovine apotransferrin and shown closely to resemble the S fragment obtained from the iron-saturated protein. 3. Fragments F and S are probably derived from the N- and C-terminal halves of the transferrin molecule respectively. 4. Bovine transferrin could donate iron to rabbit reticulocytes, but the monoferric fragments possessed little iron-donating ability.


A simple sensitive method, applicable to small polypeptides, for the determination of proteins in polyacrylamide gels after isoelectric focusing. A simple method is described for the determination of polypeptides and proteins in polyacrylamide gels after isoelectric focusing. Precipitates formed in trichloroacetic acid, under controlled conditions, are quantified densitometrically by measuring the proportion of light scattered. The procedure is of particular value in its applicability to smaller polypeptides, with mol.wts. of 3000-6000, which are not fixed adequately in gels by other procedures currently in use. The working range, over which polypeptide concentration is proportional to the effective absorbance, is approx. 1-30 microgram per component.


Lymphopenia in systemic lupus erythematosus. Clinical, diagnostic, and prognostic significance. One hundred fifty-eight patients with active, untreated systemic lupus erythematosus (SLE) were studied from the time of diagnosis. Lymphopenia was present in 75%, and another 18% of those patients developed lymphopenia subsequent to disease reactivation. Lymphopenia of less than 1500 cells/microliter occurred more frequently than any of the preliminary criteria for the classification of SLE, and it was the most prevalent initial laboratory abnormality. Lymphocyte counts were significantly lower in lupus than in the other connective tissue diseases except mixed connective tissue disease and polymyositis.


Occurrence and characterization of stable intermediate state(s) in the unfolding of ovomucoid by guanidine hydrochloride. Reversible unfolding of ovomucoid by guanidine hydrochloride, as followed by viscosity and difference-spectral measurements at 25 degrees C, pH6, occurred in two distinct steps involving at least three major conformational states, namely the native, intermediate and completely denatured states, occurring respectively in 60mm-sodium phosphate buffer, 3.5m-guanidine hydrochloride and 6m-guanidine hydrochloride. The overall native conformation of ovomucoid, as indicated by its intrinsic viscosity (5.24ml/g) and gel-filtration behaviour, differs significantly from that of a typical globular protein. Exposures of tyrosine residues in native ovomucoid measured by difference spectroscopy following perturbation with glycerol, ethylene glycol and dimethyl sulphoxide were, respectively, 0.42, 0.56 and 0.57. Of the exposed phenolic groups only one titrated normally (pK(int.), 9.91, electrostatic-interaction factor, w, 0.04). Results on difference spectra, solvent perturbation, phenolic titration and intrinsic viscosity (7.4ml/g) taken together showed that, although ovomucoid in 3.5m-guanidine hydrochloride was significantly unfolded, it retained a degree of native structure, removable with 6m-guanidine hydrochloride. In the latter, all the six tyrosine residues were available for titration, and the intrinsic viscosity of ovomucoid increased to 9.4ml/g. Furthermore, the characteristic fine structures in circular-dichrosim spectra of ovomucoid, associated with the elements of native structure, were abolished in 6m-guanidine hydrochloride, suggesting that the completely denatured state is structureless and presumably behaves as a cross-linked random coil. The latter state has been shown by analysis of the results on guanidine hydrochloride-dependence of the transition, intermediateright harpoon over left harpoondenatured, to be less stable than the intermediate state under native conditions by about 46kJ/mol at 25 degrees C. Attempts have been made to interpret the above results in the light of available information on the amino acid sequence of ovomucoid.


Relationshipp of HLA-Dw3 and HLA-B8 to Sjögren's syndrome. Nineteen patients with Sjögren's syndrome were evaluated for the presence of HLA-B8 and HLA-Dw3. HLA-B8 was found in 57.8% of patients and 20.1% of 96 controls (P less than 0.001). HLA-Dw3 was detected in 73.7% of the patients and 24.0% of controls (P less than 0.0001). Statistical evaluation of the association of both antigens with Sjögren's syndrome revealed that the primary association was with HLA-Dw3 (P less than 0.005). Patients with HLA-Dw3 had a lower mean parotid salivary flow rate (0.8 +/- 0.3 ml/10 minutes/gland versus 5.6 +/- 1.2 ml/10 minutes/gland, P less than 0.0004) and a more intense lymphocytic infiltration into labial salivary glands (focus score 7.2 +/- 0.9 versus 3.4 +/- 0.9, P less than 0.04) than did patients without HLA-Dw3. The difference in the intensity of the lymphocytic infiltration was even more significant when patients with both HLA-Dw3 and HLA-B8 were compared with patients with neither antigen (focus score 7.5 +/- 1.0 versus 2.8 +/- 0.8, P less than 0.02). In addition, all patients with severe keratoconjunctivitis sicca had both antigens. Our observations suggest that a number of genes may interact to determine susceptibility and severity in Sjögren's syndrome.


Fibrinogen turnover in progressive systemic sclerosis. Large amounts of fibrin are seen in the intima of the renal arterioles in progressive systemic sclerosis (scleroderma). The half-life or half disappearance time of plasma fibrinogen in 15 patients with scleroderma was studied using 125I fibrinogen to find whether there is an increased turnover of plasma fibrinogen paralleling this morphologic abnormality. Patients had a more rapid fibrinogen turnover than normal controls (60.7 versus 90.6 hours); the subgroup of patients with "progressive" scleroderma had a more rapid fibrinogen half-life than those with "stable" scleroderma (56.5 versus 73.2 hours). The mean fibrinogen half-life of 8 patients given intravenous heparin increased to within one standard deviation of normal, a finding that suggested that the fibrinogen molecule in these patients was capable of normal survival. There was a considerable variation of normal survival. There was a considerable variation of fibrinogen half-lives in individual scleroderma patients over time (not seen in the normal controls) which may be the result of intermittently increased fibrinogen consumption.


Rapid assessment of 99mTc-pertechnetate uptake in the knee joint as a parameter of inflammatory activity. Shortly after intravenous injection of 99mTechnetium-pertechnetate (99mTcO4-)(200 muCi), the activity of the knee joints was studied in patients with clinical arthritis of one or both knees and compared with that in normal subjects. A rest period directly before the study improved reproducibility. The values found shortly after injection were comparable with measurements at the maximum. The significance of the 99mTcO4- uptake in comparison with the histologic findings in cases of doubtful arthritis was examined.


Immunoglobulin in tophi and on the surface of monosodium urate crystals. By means of immunohistologic techniques IgG, IgM, and to a lesser degree, IgA were found in the intercrystalline matrix of gouty tophi. Smaller amounts of IgG and IgM were inconstantly found in association with tissue deposits of calcium pyrophosphate. When a technique of immune agglutination was used, the affinity of all three major classes of immunoglobulin to the surface of monosodium urate (MSU) crystals was demonstrated. Immunoglobulin and fibrin were found on MSU aspirated from large tophi.


Non-giant cell temporal arteritis. Three cases and a review of the literature. We report two cases of polyarteritis and some of hypersensitivity angiitis diagnosed by temporal artery biopsy. Autopsies showed no evidence of giant cell arteritis. A review of the English literature provides seven further autopsy cases of necrotizing vasculitis diagnosed by temporal artery biopsy. The term temporal arteritis includes many types of vasculitis, giant cell arteritis being one of them.


Idiopathic multicentric osteolysis. Report of an affected father and son. Genetic, rheumatologic, immunologic, metabolic, and renal studies of a father and son with idiopathic multicentric osteolysis are reported. The disorder appeared through mutation. The father developed symptoms as an infant, his son at age 4 years and 9 months. Both have micrognathia and hypotelorism and were exceptionally tall during the symptomatic phase of their disease. Biopsies of the son's wrist showed normal synovium, encroachment on cartilage by fibrocellular tissue, and both osteoclastic resorption and repair of affected bone. Hydroxyproline in his urine was increased. No immunologic, renal, or other metabolic abnormalities were identified.


Ankylosing spondylitis with subsequent development of rheumatoid arthritis, Sjögren's syndrome, and rheumatoid vasculitis. Twelve years after the onset of ankylosing spondylitis, a patient developed severe seropositive rheumatoid arthritis with subcutaneous nodules. This was subsequently complicated by Sjögren's syndrome and rheumatoid vasculitis. The literature is reviewed, and the nature of the association between ankylosing spondylitis and rheumatoid arthritis is discussed.


Effect of garlic oil in experimental cholesterol atherosclerosis. Addition of cholesterol in the diet of male albino rabbits produced hypercholesterolaemia, increased tissue cholesterol, and atheromatous changes in the aorta. Supplementation of garlic oil along with cholesterol significantly inhibited the hypercholesterolaemia, decreased tissue cholesterol and minimised the atheromatous changes in the aorta. These results show that the active constituent(s) in garlic responsible for its anti-atherogenic action is present in the oily fraction of garlic.


Compounds that increase oxygen diffusion in plasma. The rate of diffusion of oxygen in blood plasma is increased by adding certain compounds to plasma at very low concentration levels. Decreased oxygen diffusion rates in plasma causes local hypoxic conditions and this may play an important part in the pathogenesis of atherosclerosis.


Studies on the occurrence of circulating immune complexes in vascular diseases. The presence of circulating immune complexes was studied in 347 samples of serum from 212 patients with various vascular diseases. Two quantitative methods (complement-consumption assay and C1q-solubility test) were used for the measurement of the concentration of the complexes. Immune complexes were detected in each group of patients tested (coronary arteriosclerosis, myocardial infarction, cerebral artery sclerosis, arteriosclerosis obliterans, phlebothrombosis, pulmonary infarction). A high proportion of positivity was recorded in myocardial infarction (in 43 patients out of the 94 tested) and in arteriosclerosis obliterans (7 out of 11 cases). The possible pathogenic role of the circulating immune complexes is discussed.


Connective tissue components in normal and atherosclerotic human coronary arteries. Human coronary arteries with various degrees of atherosclerosis were analyzed for the concentration of different types of glycosaminoglycans (GAGs). The changes in GAGs were considered against the background of macroscopic atherosclerosis, and the concentration of glycoprotein-bound hexosamines, collagen, calcium and cholesterol. The concentration of calcium was increased and that of hyaluronic acid decreased even in mildly atherosclerotic coronary arteries. The additional changes in advanced atherosclerosis included the increase of collagen and dermatan sulphate and the decrease of heparan sulphate. Cholesterol was increased in mild, and even further in advanced, atheroslcerosis. The concentrations of chondroitin sulphates and glycoprotein-bound hexosamines were not significantly affected by atherosclerosis.


The in vitro inhibition of bovine milk lipoprotein lipase by a glycoprotein preparation from human atherosclerotic intima. Intimal tissue from human atherosclerotic aortae was collected by the Dermatome procedure. The tissue was extraved with 5 mM Tris.HCl buffer containing 0.3 M NaCl and 1 mM EDTA, pH 7.4. The ammonium sulfate precipitate between 0.4--0.8 saturation obtained from the extract was fractionated on a DEAE-cellulose column and the effluent was monitored for lipoprotein lipase inhibition employing purified bovine milk enzyme. The substrate used was an emulsion of purified olive oil and tritiated triolein. Human serum was the source of activator of the substrate. A peak of inhibitory activity was eluted between 0.15--0.17 M NaCl. The major component in the purified material had properties similar to a glycoprotein (lipolipin) which has previously been purified from porcine aorta and shown to inhibit lipoprotein lipase activity. The partially purified human inhibitor decreased both the basal and the serum-stimulated activity of milk lipoprotein lipase. The  inhibition was non-competitive with respect to serum. However, high levels of triglyceride substrate appeared to relieve the inhibitory effect. It is postulated that the inhibitor may be involved in an interaction with the emulsified lipid denying the enzyme access to its substrate.


Fibrinogenemia, determined immunonephelometrically, as a possible parameter in the evaluation of peripheral arteriosclerotic arteriopathy. The concentration of plasma fibrinogen increases with age and is particularly elevated in subjects with vascular changes due to atherosclerosis. Fibrinogen, which is the meeting point of the "cascade" of the coagulation factors and an important regulating factor of plasma viscosity, occupies a key position in atherosclerotic vascular pathology. This indicates the necessity to keep its value within normal limits, to avoid the clinical development of the disease or improve its evolution. Thus, frequent checks of the fibrinogenemia are necessary at various ages even if this means a big organizational burden. However, this checking can be facilitated by using automatic techniques, such as the immunonephelometric method, with which it is possible to carry out a large number of determinations in a short time.


A xerographic method for the quantitative assessment of atherosclerotic lesions. A new method has been developed for the quantitative assessment of atherosclerotic lesions, in which a conventional xerox machine is the sole equipment used. As an example, the aorta from a cholesterol-fed rabbit was removed in toto, opened longitudinally along its anterior margin, flattened in a transparent polypropylene bag and then xeroxed. The outlines of the atherosclerotic lesions were traced on translucent graph paper ruled into 1 mm squares. The squares within those outlines were counted and expressed as a percentage of the whole surface. This xerographic method is rapid and reproducible and is achieved without changing the magnification; it gives comparable or better results than other staining methods. As the aortic specimen is undamaged by fixation or staining, the specimen can be used for subsequent morphological or biochemical studies. Thus, the degree of surface involvement can be correlated with other morphological or biochemical changes in the same arterial samples.


A quantitative study of the development of sudanophilic lesions in the aorta of rabbits fed a low-cholesterol diet for up to six months. Eighteen New Zealand white rabbits were placed on a diet containing 0.25% cholesterol and 6% corn oil for periods of up to 6 months. The animals were divided into 4 gours and sacrificed after 2, 3, 4, and 6 months. The aortae were removed, stained with Sudan III, and analyzed with the polar coordinate technique. While the periorificial lesions developed more slowly on this diet than on the diet with 2% cholesterol and 6% corn oil we had used previously, there was no differences in either the location or the shape of the lesions. In the descending thoracic aorta, lesions developed initially distal  to orifices; however, significant lateral and proximal components were observed as atherogenesis progressed. The coronary lesions completely surrounded the ostia in all stages of development. The total area of the lesions was more related to time (r = 0.74, P less than 0.01), than to serum cholesterol (r - 0.51, P less than 0.05) or to cholesterol-time product (r = 0.69, P less than 0.01).


Afferents to brain stem nuclei (brain stem raphe, nucleus reticularis pontis caudalis and nucleus gigantocellularis) in the rat as demonstrated by microiontophoretically applied horseradish peroxidase. Using a retrograde tracer technique with microiontophoretically applied horseradish peroxidase (HRP), afferent projections to the brain stem raphe nuclei (BR, raphe magnus, pallidus and obscurus) and to two adjacent reticular nuclei, nucleus reticularis pontis caudalis (nRPC) and nucleus gigantocellularis (nGC) were identified. The most striking difference between the afferent projections to the BR and the adjacent nuclei as determined by this method is that afferents to the BR originate primarily from structures rostral to the pons, especially the mesencephalic central gray and the dorsal and ventral tegmentum. In contrast, the two reticular nuclei studied (nGC and nRPC) received afferent projections within or caudal to the pons-medulla. For example, the nGC receives prominent afferent projections from the gray matter of the spinal cord. In addition, evidence for interconnections between all of the adjacent nuclei (BR, nGC and nRPC) was found. Such afferent projections are compatible with the notion that the brain stem raphe nuclei may serve as connections within the brain stem for a descending system, while the nGC may be a relay in a feedback loop between the spinal cord and the reticular formation.


Excitation and inhibition of neurons in the trigeminal nucleus caudalis following periaqueductal gray stimulation. Electrical stimulation (3-4 shocks, 300 Hz, 30-150 microamperemeter) of the periaqueductal gray matter (CG) or dorsal raphé nucleus (DR) of decerebrate cats reduced or abolished the jaw-opening reflex response evoked by stimulation of either the tooth pulp or infraorbital nerve. In addition, CG or DR stimulation inhibited the response of 12 out of 16 trigeminal nucleus caudalis neurons to activation of their sensory afferent inputs. Ten other neurons recorded in the same sites, and often at the same time, but which did not respond to the sensory inputs utilized, were excited by identical stimuli to CG or DR. This excitatory response was blocked by intravenously administered naloxone (0.1-0.2 mg/kg). It is suggested that those neurons which are excited by CG and DR stimulation may be interneurons involved in pre- and post-synaptic inhibition of sensory transmission during stimulus-produced or narcotic analgesia.


Tumor membrane lymphocyte stimulation assay in patients with renal cell carcinoma. Peripheral blood lymphocytes from 15 patients with hypernephroma were stimulated with partially purified tumor plasma membranes to incorporate [3H]thymidine. Kidney tumor and normal kidney membranes were adjusted to antigenic equivalence as determined by their ability to inhibit in the HLA 51Cr microcytotoxicity assay. Membranes from control "normal" kidney adjacent to the tumor stimulated less than did the tumor. Six of eight patients responded to autologous tumor (p less than 0.05). One patient responded to allogeneic tumor of the same histological type. The importance of statistical analyses of tumor membrane lymphocyte stimulation data is discussed in relation to the assay system. Sequential studies suggest that this assay may be useful as a guideline for the monitoring of current therapeutic regimens and future immunotherapy. The results of this assay are discussed in relation to other in vitro tumor lymphocyte stimulation assays. The limitations of this assay appear to be two: (a) it can be used only in large tumor systems where there is adequate tissue for analysis and controls; (b) it may detect nontumorous antigens or nonspecific stimulators in allogeneic studies. Further studies are needed to correlate the blastogenic response with the patient's prognosis.


Regional choline acetyltransferase activity in the guinea pig heart. Choline acetyltransferase is the enzyme that catalyzes the biosynthesis of acetylcholine, the neurotransmitter of the pre- and postganglionic parasympathetic system. To assess the extent of parasympathetic innervation, enzyme activity was measured in specialized and contractile regions throughout the guinea pig heart. Enzyme activity in the right atrial appendage was 137 nmol g(-1) hr(-1). Activity was greatest in the region of the sinoatrial node (187 nmol g(-1) hr(-1). Also, enzyme activity was high in the regions of the atrioventricular node (153 nmol g(-1) hr(-1), the proximal conduction bundles (133 nmol g(-1) hr(-1), and the base of the anterior papillary muscle of the right ventricle (179 nmol g(-1) hr(-1), which contains the moderator band and Purkinje fibers. In contrast, the enzyme activity in the inferior interventricular septum and the free walls of the right and left ventricles, which are more predominantly contractile tissue, was 67 +/- 6,108 +/- 14, and 56 +/- 11 nmol g(-1) hr(-1), respectively. This activity is significantly lower than in the right atrial appendage. These results suggest that the density of parasympathetic innervation is similar in all the components of the conduction system, from the sinoatrial node to Purkinje tissues. Furthermore, the parasympathetic innervation of regions specialized for conduction is up to four times more dense than that of contractile regions.


Serum chromium and angiographically determined coronary artery disease. Human aortas sampled from populations where there is little advanced atheromatous plaque formation contain higher concentrations of chromium than do aortas from populations in which atheromatosis is prevalent. In the present study serum cholesterol, triacylglycerols, and chromium (Cr3+) concentrations were measured in 32 subjects in whom coronary artery disease was assessed by cineangiography. The distribution of subjects with diseased and normal arteries overlapped below 5.50 microgram of chromium per liter. Only subjects free of coronary artery disease had chromium concentrations greater than or equal to 5.50 microgram/liter. The role of chromium was assessed in the context of the selected risk factors: cholesterol, triacylglycerols, and systolic and diastolic blood pressure. The group with coronary artery disease had significantly lower serum chromium concentrations than did the group with normally patent arteries.


The structure of hemoglobin Hopkins-2.

The structure of hemoglobin Hopkins-2. Hemoglobin Hopkins-2 was originally reported to contain two abnormal alpha chains. Both had aspartic acid instead of the normal histidine at position alpha 112; the second alpha chain also contained two additional amino acid substitutions. In subsequent reports of the structure of this hemoglobin, only the His replaced by Asp substitution was found. To determine whether the second abnormal alpha chain was limited to the original kindred, the structure of their hemoglobin was redetermined. Only the His replaced by Asp substitution was present.


The fine structure of ventricular cells in the brains of mouse embryos homozygous for the loop-tail gene. The neural tube in normal (+/+), heterozygous (Lp/+), and abnormal (Lp/Lp) mutant mouse embryos ranging in age from 10 to 12 days of gestation was studied by means of transmission electron microscopy. In the abnormal embryos, ventricular cells in defective regions of the brain show distortions and crowding together of internal cellular processes and a decrease in blebs and bulbous projections, as compared  with their normal counterparts. At 12 days' gestation the abnormal brains show a scarcity of the T-shaped internal cellular processes characteristic of normal brains. The abnormal brains also show increased amounts of intercellular space and extensive gaps between the cells, particularly in basal regions. There are no striking differences between the normal and abnormal brains at 10 to 12 days' gestation with respect to the appearance and distribution of cilia, microfilaments, microtubules, tight junctions, and ribosomes.


Postnatal development of the granule cell in the kitten cerebellum. Newborn to 9-week-old kittens were anesthetized and then sacrificed by inserting a scalpel into the thoracic cavity and severing the descending aorta. The cerebella were removed and processed according to Golgi-Cox and rapid Golgi modifications. The results indicate that the dendritic processes of the granule cells undergo a marked postnatal maturation even though there is little postmigratory change in the size and shape of the cell body. A club-shaped swelling of the distal portions of the dendrites is observed by the end of the first week; this "claw" becomes multilobated by three weeks and displays elaborate digitiform projections by nine weeks. The sequence of postnatal development in the kitten appears to follow a time course similar to that observed in other species. A possible correlation between the postnatal development of the granule cell and the postnatal development of the spincocerebellar system is discussed.


Successful treatment by overdrive pacing of recurrent quinidine syncope due to ventricular tachycardia. A 54 year old patient who experienced recurrent ventricular tachycardia subsequent to quinidine administration for conversion of atrial fibrillation is described. Over a 10 hour period, 25 sustained episodes of ventricular tachycardia occurred for which electrical cardioversion was required in addition to numerous self-terminating paroxysms. Medical therapy with lidocaine, procainamide and propranolol was unsuccessful in controlling the arrhythmia. However, placement of a transvenous right ventricular pacemaker with overdrive pacing at a rate of 110 beats/min abrupty terminated all further ventricular ectopic activity during the period of quinidine elimination. Temporary overdrive pacing may be the treatment of choice for refractory, recurrent, ventricular tachycardia associated with quinidine therapy.


Radiographic abnormalities in tubular bone after rigid plate fixation in rabbits. Rigid stainless steel osteosynthesis plates (DCP/ASIF) were attached bilaterally onto the intact tibio-fibular bone in 40 rabbits. Axial compression was used on the right side, neutral plate fixation on the left. The radiographic appearances of the bone were analysed 1 day, and 1, 3, 12, 24 and 36 weeks postoperatively. The changes in the bone were progressive, related to the time of fixation, and similar in the right and left leg. The main alterations were a progressive thinning of the cortical bone under the plate and a progressive cancellous transformation of the tubular bone.


Medical adrenalectomy with aminoglutethimide: clinical studies in postmenopausal patients with metastatic breast carcinoma. The use of adrenalectomy and hypophysectomy in the management of postmenopausal patients with metastatic breast carcinoma is reserved for highly selected patients. As an alternate approach, a pharmacologic method of inhibiting adrenal cortical secretion was developed which consisted of the daily administration of 1000 mg of aminoglutethimide to block steroidogensis and either dexamethasone (2.0-3.0 mg/day) or hydrocortisone (40-60 mg/day) as replacement glucocorticoid. This regimen markedly suppressed plasma levels of DHA-S, androstenedione, estrone, and estradiol, and urinary levels of aldosterone. Of 50 patients treated, 19 (38%) demonstrated either a complete (8/19) or a partial (11/19) objective disease remission which lasted for 18.05 +/- 3.1 months (mean +/- SEM). In 10 (20%) patients, there was stabilization of disease (7.8 +/- 1.2 months), accompanied by symptomatic relief of bone pain in six (12%). There was disease progression in 20 (40%) patients. The acute side effects of aminoglutethimide therapy were significant and consisted of transient lethargy (41.5%) and a cutaneous rash (35.8%). Chronic toxicity was negligible. The medical adrenalectomy regimen of aminoglutethimide plus glucocorticoid offers a suitable alternative to surgical adrenalectomy or hypophysectomy in the management of postmenopausal patients with metastatic breast carcinoma.


Anterior border of the left atrium on conventional heart films. The appearance of the anterior left atrial wall on conventional lateral chest films is presented. The position of this structure is often marked by fatty tissue surrounding the posterior wall of the ascending aorta. A markedly enlarged left atrium may overlap the aorta. The delineation of the anterior border is still sometimes possible by observing a layer of epicardial fat surrounding the atrial wall or the interatrial sulcus. The importance of short exposure time as well as the use of the findings in practical diagnostic work is discussed.


[Enteritis and its complications. Etiology and most frequent anatomoclinical picture]. A study is made of the environmental, etiologic, pathogenic, clinical and anatomical characteristics of the bacterial or parasitic enteral processes which most frequently cause death to the patients. Considerations on the pathogenesis of the complications found at the postmortem studies in children with diarrhea are likewise offered.


[Intrauterine infection in relation to the frequence of prematurity and congenital malformations]. In order to find the relationship between intrauterine infection with frequency of prematurity and of congenital malformations, 1,016 newborns were studied and were divided into four groups: 324 normal babies, 606 prematures, 44 with malformations and 42 infected patients; the latter were confirmed through titration of serum antibodies. The first index of infection was the concentration of IgM greater than or equal to 28 mg. %; thus, the first groups showed evidence of infection in 0.9%; the second in 11%; the third in 40.9% and the fourth, in 100%.


[Skin response to phytohemagglutinin in patients with typhoid fever, bronchial asthma and Hodgkin's disease]. The skin response to phytohemagglutinin was investigated as an index of cellular immunity in group of 50 children made up of 11 with thypoid fever, 13 with bronchial asthma type I, 15 with Hodgkin's disease and 11 controls. A mild but statistically significant (p less than 0.005) depression was found in the skin response of patients with typhoid fever and moderate in patients with Hodgkin's disease. In patients with bronchial asthma, it was similar to control cases; thus, in the future, finer parameters are considered necessary in these cases to investigate cellular immunity.


[Polyradiculoneuritis. Increase of the occurrence of the severe forms]. The clinical histories of 97 cases admitted from 1963 to June 1977 with the diagnosis of polyradiculoneuritis were reviewed. A change in the epidemiology related with an increase in frequency of severe forms that reached up to 50.0% during the last 3 years, against 15% for the previous years, was observed. Overall lethality was 4.1%. Permanent sequelae resulted in 18.3% of the patients and the greatest contribution to this came from the group of severe forms, since 53.3% of them was definitely affected.


[Neonatal infection and high IgM at birth]. To determine the concentration of IgM to be used as intrauterine infection indicator, the sera of 2,906 newborns were studied. Five hundred and sixty-three (19.4%), showed high concentrations of IgM and out of 199 pair sera studied, serologic evidence of infection was found in 21.1%. Out of these cases, 80.9% showed no clinical disturbances during their first two months of life. The agents most frequently found responsible for intrauterine infection were cytomegalovirus, influenza A2 H-K, influenza B, parotiditis and rubella. Based on this study, we may conclude that determinations of high concentrations of IgM with the technique used is still a good index to detect intrauterine infections, specially considering that the clinical manifestations of disease may be minimal or may pass unnoticed during the first days of life and that by no means should this test be done instead of other clinical or laboratory studies to investigate such questions.


[Osteomyelitis in the newborn].

[Osteomyelitis in the newborn]. The study included 17 cases of osteomyelitis in the newborn period, admitted through a period of 10 years. Males showed a rate of 1.8:1 over females. The possible port of entry was established in 11 cases. The most important clinical signs were: edema, erythema, hypomotility and fluctuation. In one third of the cases, systemic infection signs were eident and in 4 cases, confirmation resulted from blood cultures positive to gram-negative germs. The germ most frequently isolated in the secretion was S. aureus, followed by Klebsiella. The analysis included the pathogenesis that explains the high percentage of articular participation (65%) and the presence of multiple foci which, together with other characteristics, make osteomyelitis at this age, different from that seen during other stages of life in terms of diagnosis, management and prognosis.


[Supraventricular paroxysmal tachycardia as a pediatric cardiologic emergency]. Seven cases with the diagnosis of supraventricular paroxysmal tachycardia, confirmed by the electrocardiogram were included in the study. Their ages fluctuated from 19 days to 14 years, with predominance of small infants. The etiology was varied. They were managed with digitalis and conversion to sinusal rhythm in a short term. The drug was maintained to prevent relapses and/or heart failure. No mortality issued. We recommend the use of digitalis as selective drug because it may be readily obtained and its appication and response are sure. The clinical findings and results agree with the literature reviewed.


[Juvenile rheumatoid arthritis. Study of 46 cases]. Forty-six children with the diagnosis of juvenile rheumatoid arthritis (JRA) were studied according to the criterion of the American Rheumatism Association (ARA). No predominance was seen in terms of sex. Preschool age was preferably affected and the poliarticular form was the most frequent at the initiation. Sixty per cent of the cases followed a recurrent course. The routine clinical manifestations were fever, arthralgia, visceromegalia and exanthema. The knees were found to be the joints most frequently affected in one half of the cases. The laboratory tests showed in all of the cases, increased sedimentation rate, hypergammaglobulinemia and in 33% of the cases, the rheumatoid factor was positive. The most frequent X-ray changes were: increase of soft parts and osteoporosis in 38% of the cases, without radiological changes in 25% of them. Thirty-four percent of the patients remained without articular sequelae and approximately in one half of the cases, functional incapacity was minimal.


[Long term mental consequences in children with purulent meningoencephalitis]. Long-term intellectual sequelae were studied in 14 children who suffered purulent meningitis during their first two years of life. A proportion of 50.0% showed an I.Q. under 90 points, but no statistical difference was found when compared with a control group made up with siblings of the patients. The main difference between both groups corresponded to a greater percentage of inadequate development in patients who had suffered meningitis. This disturbance is chiefly related with environmental factors that showed the existence of familial and social integration problems in 86% of children having an I.Q. under 90 points, against only 14% of cases showing an I.Q. above 90 points.


[Postmortem examination of patients with non-surgically treated ruptured aneurysms (author's transl)]. The is no doubt that radical surgery has been successful in preserving the lives of patients with ruptured aneurysms. On the other hand, no matter how successful we have been we cannot and must not forget the patients who died without undergoing surgery. Therefore, we investigated from clinical course, laboratory work, including angiography and autopsy, 38 cases who died during hospitalization in order to elucidate the causes of death in the natural course and to find the possibilities of minnimizing the number of non-operative fatalies. We classified the causes of death into 3 groups: intracranial hemmatomas, brain ischemia due to vasospasm, and aggravation of the general condition. From the study, it was concluded that most of the lives could have been saved, if appropriate diagnosis and treatment had been made as soon as possible after the initial hemorrhage.


[Analysis of urine in the child without renal disease and with fever]. The study comprised 24 patients with ages between 5 months and 13 years and fever from different extrarenal infectious diseases. Seventy-nine samples of urine were taken to determine proteins, glucose, ketone, hemoglobine and study of the urinary sediment. Electrophoresis of proteins was practiced to samples with higher content of them. Considering a normal rate of protein the figure above 4 mg/hr/m2, 70% of the patients and 39% of samples showed abnormality with figures up to 52 mg/hr/m2, or 1,259 mg/l. All of them excreted albumin and alphaglobulins; 15, betaglobulins and 4, gammaglobulins. No relationship was found between the amount or type of protein excreted and the degree of fever, age of the patient or type of disease. Thirty-seven per cent of the patients showed ketonuria, 29% erythrocyturia, 21% hemoglobinuria, 16% glycosuria, 12% cylindruria, and 8% leukocyturia. The duration of abnormalities lasted from 1 to 4 days and the magnitude was usually mild, having obtained in all of the cases, a normal sample of urine and a negative urine culture before discharge of the patient. Findings with other studies are compared and the pathogenesis of abnormalities is discussed.


[A case of cerebral vascular disease associated with prosopagnosia and minor hemisphere syndrome (author's transl)]. A case of "apractognosia", a syndrome of minor hemisphere, and prosopagnosia was described. A 68 years old, right handed, hypertensive man who had experienced an episode of left hemiparesis of several weeks' duration developed a syndrome of minor hemisphere, metamorphopsia, visuoconstructive disability, spatial dysgraphia, topographical and geographical disorientation and dressing apraxia, with associated left homonymous hemianopsia. A right occipital lesion was disclosed by an isotopic brain scan and a CT scan. The CT scan also disclosed a generalized cerebral atrophy. Memory disturbance, both verbal and visual, was observed. Verbal memory was examined by coupled words test, digit span and 10-words learning test, and visual memory by spatial pointing span and memorizing unfamiliar faces. The patient was able to differentiate unfamiliar faces, but was not able to memorize them correctly. Although the patient has complained of a disturbance of recognizing familiar faces (prosopagnosia), but actually, sometimes he was able to recognize the members of his family by the faces and sometimes he was not. He has stated that he could not recall familiar faces. The authors suggested prosopagnosia is, as Benton (1975) has supposed, considered to be the limited disorders of memory and association.


[Intracranial pressure in patients with diffuse cerebral arterial spasm following ruptured intracranial aneurysms (author's transl)]. Intracranial pressure (ICP) was recorded continuously for 2 to days in 12 pre-operative patients with angiographic evidence of diffuse cerebral arterial spasm due to ruptured intracranial aneurysm. The ICP pattern of the low level (below 15 mmHg) and flat type was registered in the early stage of the arterial spasm in 11 patients in whom the ICP recording was made within 11 days after the hemorrhagic attack. During the period of low and flat ICP pattern, 6 patients showed little or no neurological deterioration whereas 5 patient showed impaired consciousness or neurological deficits. In 9 out 11 patients, the low level of ICP was followed by an abnormal deterioration. Continuous ventricular drainage was performed to control the secondary increased ICP in 7 patients and they survived, 4 of them with good results but remaining 3 with severe neurological deficits.


Detection of abnormal left ventricular wall movement during isovolumic contraction and early relaxation. Comparison of echo- and angiocardiography. Abnormal left ventricular wall movement during isovolumic contraction and early relaxation was assessed from simultaneous apex and echocardiograms in 50 patients with ischaemic heart disease, and compared with estimates from the corresponding digitised left ventriculograms. During isovolumic contraction, a normal angiogram was accompanied by normal apex-dimension relations in 13 out of 14 cases. In 19 cases, there was angiographic evidence of discrete outward wall movement during isovolumic contraction which was associated with abnormal apex-dimension relations in 15. During isovolumic relaxation, of 14 cases who were normal angiographically, apex-dimension relations were normal also in 11, which in 36 patients with abnormal wall movement on angiogram, apex-dimension relations were abnormal in 30. Correlation was less good between echocardiographic and angiographic estimates of left ventricular minor dimension (r = 0.75), and was absent between those of peak rates of dimension change during systole and diastole. Asynchronous onset of inward wall movement and the distribution of regional abnormalities of overall wall movement amplitude were unrelated to apex-dimension relations. The apex-dimension relation is thus a sensitive and specific means of detecting abnormalities of left ventricular wall movement during isovolumic contraction and early relaxation, unaffected by other manifestation


Echocardiographic analysis of posterior mitral leaflet movement in mitral stenosis. Echocardiographic analysis of the movement of the posterior mitral valve leaflet in 60 patients with lone mitral stenosis, 35 patients with aortic stenosis, and 18 patients with aortic and mitral stenosis showed a spectrum of initial posterior mitral valve leaflet movement in early diastole. The classical anterior movement was seen in 36 out of 60 patients with mitral stenosis (60%), and 8 out of 16 patients with aortic and mitral stenosis (50%). Normal posterior movement was present in all patients with lone aortic stenosis but was also seen in 10 patients (17%) with mitral stenosis and 6 patients (33%) with aortic and mitral stenosis. The remaining patients with mitral stenosis or aortic stenosis and mitral stenosis showed a biphasic type of initial movement. Patients with anterior movement had a mean calculated mitral valve area from cardiac catheterisation significantly smaller than the rest (P less than 0.001), but neither biphasic nor posterior movement excluded severe mitral stenosis. The distinction between patients with mitral stenosis and initial movement of the posterior mitral valve leaflet and patients with left ventricular discompliance is possible when there is sinus rhythm. Late diastolic anterior movement of the posterior mitral valve leaflet during atrial contraction is diagnostic of true mitral stenosis.


Echocardiographic identification of aorta and main pulmonary artery in complete transposition. No completely reliable echocardiographic technique has been described for the separate identification of the aorta and main pulmonary artery in complete transposition of the great arteries. A mechanical wide-angle (60 degrees) sector scanner has been applied to this problem in 17 infants and young children, including 8 newborns before angiocardiography. In all patients a longitudinal scan (saggital section) identified the main pulmonary artery by its directly posterior course immediately beyond the pulmonary valve, and the aorta by its retrosternal course upwards before turning posteriorly above the main pulmonary artery. In addition, a high transverse scan showed the precise spatial relation of the great arteries, and, in 11 of the 17, tilting the scanning plane upwards showed branching of the main pulmonary artery. In 8 infants examined with M-mode echocardiography, an upward sweep from the pulmonary valve showed abrupt termination of the echo from the posterior wall coinciding with the posterior arching of the main pulmonary artery.


Natural history and evaluation of Q waves during acute myocardial infarction. Serial 72 point praecordial electrocardiographic maps were recorded in 45 patients with uncomplicated acute anterior myocardial infarction. These were analysed and the serial changes in Q waves and ST segments were recorded. Cardiospecific enzyme release curves were constructed from repeated measurements of the plasma activity of the myocardial isoenzyme of creatine kinase (MB CK) during 4 days after the onset of chest pain. The praecordial maps showed that Q waves appeared in the second hour after the onset of chest pain and the development of this electrocardiographic sign was completed within 12 hours. There were complex relations between the development of Q waves, the natural history of ST segment elevation, and the release of MB CK activity. The praecordial area of ST segment elevation at 2 hours was directly related to the fully developed Q wave area at 24 hours after the onset of chest pain. The electrical activity of affected myocardium was lost before peak plasma MB CK activity.


Coronary artery ectasia--a variant of occlusive coronary arteriosclerosis. In a study of 1000 consecutive coronary arteriograms, 12 patients (all men) had coronary artery ectasia. Ectasia was found most frequently in the circumflex or right coronary artery. Only 1 patient had ectasia in the left anterior descending coronary artery. In 11 patients, ectasia of one artery was associated with severe stenosis or occlusion of other vessels, typical of arteriosclerosis. Histology from an ectatic segment in one of this group showed changes of severe arteriosclerosis with extensive intimal fibrosis and destruction of the media. One patient had a mixed collagen vascular disease. Measurement of coronary sinus flow in 2 patients with coronary artery ectasia showed flows in the range of patients with non-ectatic coronary artery disease. At cardiac surgery flows down the graft to ectatic arteries were in the same range as in grafts to non-ectatic vessels. Patients with coronary artery ectasia should be anticoagulated.


Height and weight gain after total correction of Fallot's tetralogy. In a consecutive series of 100 cases of Fallot's tetralogy undergoing total correction, the mean birthweight was found to be just below the 50th centile. Blalock-Taussig or Waterston shunts were performed in 45 patients. Before their shunt operations a significant number of patients had become retarded in height and weight development but after the shunt operations many patients showed accelerated height and weight gain. Of the 73 patients followed up 2 years after correction, 23 had experienced accelerated development in both weight and height, but 11 patients remained on or below the 3rd centile for weight and height. In terms of the age at shunting, the oxygen saturation immediately before total correction, the age at total correction, the number of ventriculotomy patch repairs, and the post-correction ventricular pressure ratios, no significant difference could be detected between those 23 patients who had undergone accelerated development and those 11 who had remained retarded. More shunts, particularly of the Waterston type, were performed in the retarded group.


Torsade de pointes ventricular tachycardia in a newborn infant. A case of paroxysmal ventricular tachycardia of torsade de pointes variety occurring in a newborn infant is described. A rare problem in the newborn, ventricular tachycardia has been associated with congenital heart disease, electrolyte abnormality, and cardiac tumour. In this case, the association was with myocarditis. The arrhythmia was refractory to treatment, and the infant died.


Diarrhoea induced by migration of a pacemaker generator. A case of intraperitoneal migration of a pacemaker generator is described. Chronic diarrhoea and abdominal discomfort were relieved by its removal.


Equations for vapour pressure versus temperature: derivation and use of the Antoine equation on a hand-held programmable calculator. The Antoine equation is a semi-empirical equation which expresses vapour pressure as a function of temperature. A new, rapid and highly accurate method for obtaining its three constants from experimental data is presented and applied to ethanol, water and 14 anaesthetic substances. Alternative vapour pressure equations are discussed and references for original temperature--vapour pressure data are summarized. A series of equations utilizing vapour pressure is detailed: these formulas are of use in both the practice and teaching of anaesthesia.


A vaporizing system for programmed anaesthesia. A vaporizing system for closed circuit "programmed" anaesthesia is described. Despite its location within the circuit, the vaporizer controls directly the input of volatile anaesthetic agents irrespective of fluctuations in ventilation of the anaesthetized subject. It is interfaced easily with electronic controllers and has an accuracy approaching 1.0% under laboratory conditions. During experimental anaesthesia, it maintained stable end-tidial concentrations of halothane at 1.2 MAC (the intended value) despite wide variations in ventilation.


Bedside estimation of whole blood lactate. A new instrument for the measurement of lactate in biological fluids, the Lactate Analyzer 640, has been evaluated. The method of use recommended by the manufacturer for blood samples was found to be inadequate. A new method of sample preparation for the instrument, based upon immediate haemolysis and fluoridation of blood, has been developed, allowing measurement of whole blood lactate concentration to be performed on samples as small as 150 microlitre within 5 min of withdrawal. The instrument is designed to be operated by a medical practitioner. Excellent correlation with a conventional enzymatic assay was found. These features make this new method particularly applicable to rapidly changing clinical situations such as shock in adult patients, asphyxia neonatorum and intrapartum foetal hypoxia.


Preliminary investigation into a new method of assessing the quality of anaesthesia: the cardiovascular response to a measured noxious stimulus. It is suggested that the autonomic response to noxious stimulation may be measured during anaesthesia. The response may be marked despite considerable depression of cortical activity as measured by the cerebral function monitor. Preliminary results suggest that large doses of narcotic analgesics reduce the autonomic response to noxious stimulation and this may be used as a test to compare the potency and duration of action of narcotic drugs during anaesthesia. It is possible that the measurement of beat-to-beat variation in heart rate may be used to monitor autonomic nervous system activity and, indirectly, the degree of stress during surgery, provided that the efferent limb of the reflex arc remains intact. With the aid of the cerebral function monitor and with an index of reflex response to noxious stimulation it should be possible to monitor the effect of anaesthesia on the central nervous system more precisely.


Haemodynamic responses to induced arterial hypotension in children. Cardiovascular measurements were made in 12 children in whom arterial hypotension was induced with pentolinium and halothane. Propranolol was given to five patients who exhibited tachycardia. Measurements were made at the following stages: before induction of hypotension, 5 min after the administration of pentolinium, when a "dry" opening field was obtained, at the time of maximum hypotension and when arterial pressure had returned to 90% of the control values. The decrease in arterial pressure 5 min after the administration of pentolinium was accompanied by tachycardia, but there was no significant change in cardiac index and the operative field was congested. A substantial decrease in cardiac output occurred when a "dry" field was obtained.


Glucose tolerance during anaesthesia and surgery. Comparison of general and extradural anaesthesia. The effects of general and extradural anaesthesia on glucose tolerance and plasma cortisol concentration in the surgical patient were assessed. Normal glucose tolerance and insulin release were observed under extradural anaesthesia, whereas general anaesthesia produced decreases in both glucose tolerance and insulin release. The plasma cortisol concentration was increased in both groups. These results indicate possible nutritional advantages in conducting surgery under extradural anaesthesia.


Acute epiglottitis in children. Review of 27 patients. Of 27 children treated for acute epiglottitis, 26 survived. Eighteen patients treated by nasotracheal intubation were devoid of sequelae, whereas of six patients in whom tracheotomy was performed, one developed an ugly scar and two had slight tracheal stenosis at the tracheostomy site. In 22 of the 27 children the diagnosis of the referring physician was wrong, causing serious delay in securing the airway. We conclude that short-term nasotracheal intubation and antibiotic prophylaxis is the optimal treatment for acute epiglottitis in children. However, such patients are often in a critical condition, and it is essential that a well-planned procedure for examination and treatment is established in each hospital.


The fibreoptic laryngoscope in the management of cut throat injuries. The trachea of a man who had cut his own throat was successfully intubated using a fibreoptic laryngoscope after attempts at conventional oral intubation had failed. The advantages of the fibreoptic instrument and the method of use in such an injury are discussed.


Specific active immunotherapy does not prolong survival in surgically treated patients with stage IIB malignant melanoma and may promote early recurrence. A prospective trial with concurrent controls was designed to assess the effects of specific active immunotherapy in patients receiving intermittent cytotoxic chemotherapy (DTIC + Vincristine) as an adjuvant to surgery in Stage IIB malignant melanoma. The treated group received monthly irradiated allogeneic melanoma cells and BCG, and the controls BCG only. Sixteen patients in the treatment arm had a median relapse-free interval of 5 months, compared to 8 months in 12 controls given chemotherapy and BCG, and because of this we felt that continuation of the study was unjustified on ethical grounds. Although all the controls who relapsed did so at distant sites, 7/11 patients given specific active immunotherapy relapsed initially within the lymphatic drainage area of the primary tumour. The median intervals from starting treatment to relapse at distant sites, and the median survival were identical in the 2 groups. We conclude that immunotherapy comprising irradiated allogenic melanoma cells as employed in this study does not prolong survival in surgically treated Stage IIB malignant melanoma and may even promote early, local relapse.


Relationship of cell-mediated cytotoxicity against melanoma cells to prognosis in melanoma patients. The cell-mediated cytotoxicity (CMC) of blood mononuclear cells against cultured human melanoma cells was measured in patients after surgical removal of localized melanoma, at a time when they were considered on clinical grounds to be free of melanoma. It was found that the distribution of CMC values against melanoma cells in melanoma patients was different from that in control subjects, and several sub-populations of melanoma patients were evident on the basis of these measurements. No difference in distribution of CMC values was found against non-melanoma cells, which suggested the changes were specific for melanoma. The proportion of patients with recurrence of melanoma was compared between the patient groups with low, normal or high CMC values against cultured melanoma cells after surgery. Analysis for periods extending to 2 years showed that patients with low CMC values after surgery had a significantly higher incidence of recurrence from melanoma than patients with normal or high CMC values. These results suggest there may be a sub-group of melanoma patients who have intrinsically low CMC against melanoma cells, and that this may be an important predisposing factor in the development of recurrent melanoma.


Complement receptor subtypes C3b and C3d in lymphatic tissue and follicular lymphoma. To substantiate the origin of follicular (nodular) lymphoma cells from germinal-centre cells, the lymphoma cells from 7 patients with follicular lymphoma and from 9 tonsils and 2 lymph nodes were studied for the presence and distribution of complement-receptor subtypes (i.e., the receptors for C3b and C3d). It was found that erythrocytes coated with antibodies and C3d (EAC3d) adhered exclusively to germinal centres, whereas erythrocytes coated with antibodies and C3b (EAC3b) adhered to germinal centres and in many instances to the regions between them. These findings indicate that germinal-centre cells bear both complement-receptor subtypes and that the B cells of the interfollicular area, which belong at least in part to the precursors of plasma cells, bear only a receptor for C3b. In frozen sections of follicular lymphomas, a similar distribution of complement-receptor subtypes was observed; EAC3d was bound exclusively to the neoplastic nodules, and EAC3b adhered to the neoplastic nodules and adjacent paranodular tissue. Receptor studies on suspended cells of both normal tonsils and follicular lymphomas revealed a slight predominance of EAC3d(+) cells or equal numbers of EAC3b(+) and EAC3d(+) cells. The complete congruence in the expression and distribution of complement-receptor subtypes between tissues from follicular lymphomas and those from normal and hyperplastic tonsils or lymph nodes suggests that follicular lymphoma represents the neoplastic counterpart of the reactive germinal centre.


Lymphocyte defect in plasmacytoma-bearing mice. Multiple myeloma is often associated with humoral immunodepression in both man and mouse. When mice bearing the humorally immunodepressive plasmacytomas TEPC-183 and SPQC-11 were injected with SRBC, the rise of serum haemolysins was significantly less than that of non-tumour-bearing mice. Mice with the plasmacytomas MPC-11 and MOPC-315 have an antibody response similar to normal mice when injected with SRBC. Following immunization, normal mice and those bearing MPC-11 showed a 2- to 3-fold increase in total spleen lymphocytes. Mice bearing TEPC-183 or SPQC-11, the plasmacytomas causing an impaired antibody response, has significant increase in spleen lymphocytes under the same conditions. Mice bearing MOPC-315 had a very high initial count of spleen lymphocytes, which did not further increase upon immune stimulation.Incubation of lymphocytes from plasmacytoma-bearing mice with PHA did not produce an increase in TdR incorporation and in some cases even caused a decrease in TdR incorporation.Lymphocytes from mice bearing TEPC-183, SPQC-11, and MOPC-315 incorporated less TdR in response to LPS than did normal mice. On the other hand, mice bearing MPC-11 incorporated about as much TdR as did normal mice following LPS stimulation. Thus, the defect in the ability to respond to LPS in vitro correlated with the lack of an increase of spleen lymphocytes in mice bearing these tumours following antigenic stimulation in vivo.No immunodepressive properties of serum from mice with plasmacytoma could be detected.


Effects of amputation and Corynebacterium parvum on tumour metastases in mice. The effects of operation (lower-limb amputation) on the growth of the Lewis lung tumour and its metastases were studied. The role of C. parvum in counteracting these effects was investigated. Anaesthesia alone or with amputation did not affect primary tumour growth. C. parvum depressed this growth. Anaesthesia did not affect the number of pulmonary metastases, but amputation caused a significant increase. C. parvum inhibited metastases and completely counteracted the effects of operation on them. Large doses of cortisone acetate significantly increased metastases but small doses had no effect. Experiments with adrenalectomized mice suggested the effects of operation were due to non-specific stress.


Variation in response of xenografts of colo-rectal carcinoma to chemotherapy. Ten xenograft lines of human colonic and rectal carcinomas have been established in immune-suppressed mice. Mice bearing tumours in the 2nd to 6th passage were treated with maximum tolerated doses of 8 chemotherapeutic agents and tumour growth delay was estimated in terms of the number of volume doubling times gained by the treatment. The average response corresponded to a delay of only about 0.5 doubling times, but some tumour lines showed a good response to some agents. Twenty-three out of about 700 treated tumours failed to regrow. Statistical analysis showed no consistent difference in sensitivity among the tumour lines, but melphalan, 5-fluorouracil and hexamethylmelamine stood out as the most effective agents. A study of two-drug combinations showed that their order of administration had little effect on response. Only 4 of the patients who donated the xenografts were treated with chemotherapy, but among these there was some evidence that response in the xenografts correlated with response in the patient.


Repeated partial hepatectomy as a promoting stimulus for carcinogenic response of liver to nitrosamines in rats. Partial hepatectomy 24 h before a single i.p. dose of dimethylnitrosamine, diethylnitrosamine or ethylmethylnitrosamine increased the carcinogenic response in the liver of rats as determined by the number of tumours and the number of "focal proliferations" produced. Secondly, in rats given a single i.p. dose of diethylnitrosamine, 3 partial hepatectomies 5, 10 and 15 weeks after dosing the animals increased the carcinogenic response in the liver. The stimulus of repeated partial hepatectomy therefore appears to act as a "promoting agent" for liver carcinogenesis, that is if the single dose of diethylnitrosamine is regarded as an "initiating agent" in the terms of the two-stage hypothesis.


Influence of repeated liver regeneration on hepatic carcinogenesis by diethylnitrosamine in mice. In mice given a single dose of diethylnitrosamine, a hepatonecrotic dose of carbon tetrachloride, 5 weeks after dosing with DEN and repeated 6 times at 4-weekly intervals, augmented the tumour yield in the livers. A single hepatonecrotic dose of CCl4 24 h before a single dose of DEN also increased the number of tumours produced. The effect of the repeated administration of CCl4 after the dose of DEN occurred in addition to, and was therefore independent of, the enhancing effect of a single dose of CCl4 before DEN. These results may be interpreted as implying (1) that the liver in the regenerative phase after a hepatonecrotic dose of CCl4 is more susceptible to an initiating action of DEN, i.e. produces more potential foci of tumour growth than in the normal liver and (2) that the repeated doses of CCl4 leading to repeated phases of regeneration, after the dose of DEN, provide a promoting stimulus.


Ultrastructure and biological markers of neoplastic change in adult mouse epithelial cells transformed in vitro. The ultrastructure and in vitro growth properties of 5 tumorigenic mouse submandibular-gland epithelial cell lines were studied. In all lines, in vitro acinus formation occurred, and well differentiated epithelial cells showing epithelial microvilli and desmosomes and cytoplasmic tonofilaments were present. None of the cells showed specific ultratructural features of the normal differentiated submandibular-gland ducts. All the lines formed colonies in semi-solid agar and on confluent monolayers of BALB/c 3T3 cells, and all lacked density-dependent inhibition of growth, as demonstrated by a random distribution of [3H]TdR labelling throughout growing colonies. These 3 growth properties appear to be reliable in vitro markers for epithelial neoplastic transformation in this system, but colony-forming efficiency in agar is lower than that reported for many transformed mesenchymal cells.


Serial cytogenetic studies of human colonic tumour xenografts. Chromosome studies have been made of 2 human colonic tumour lines maintained as xenografts in immune-deprived mice. In both tumours human karyotypes were retained, although progressive changes occurred during serial passage. In one tumour, independent gain of a chromosome 19 was found in the stemline and 2 sidelines. In the other tumour there was selection for a sideline containing a particular deleted marker chromosome. The advantages of chromosome analysis in a xenograft system, both for the study of human solid tumour karyotypes and for monitoring the continued presence of the human genome, are discussed.


Incidence of bilateral tumours in a population-based series of breast-cancer patients. I. Two approaches to an epidemiological analysis. This paper gives the incidence in the Birmingham Regional Cancer Registry (England) of a second primary tumour in the contralateral breast among nearly 22,000 patients registered with a first primary in the breast between the years 1936 and 1964. The results, based on more than 90,000 women-years at risk and 399 second primary tumours, are presented with reference to 2 methods of analysis. In assessing risk, the principal factors investigated were age at first and second primary diagnoses and the interval between diagnoses. The results are discussed in terms of current aetiological hypotheses. On the basis of a method which included coincidental tumours, the overall risk of a tumour in the contraleteral breast was found to be 3.0 times that in the general population of a first primary. The corresponding risks for 3 main age-ranges (at the time of diagnosis of the first primary tumour) were 5.6 (ages 15-44 years), 3.7 (45--59 years) and 1.8 (60+ years). When coincidental tumours were excluded from the analysis, the relative risk was found to be 2.4 overall and 5.3, 3.0 and 1.0 for the 3 age-ranges, respectively. The level of risk was negatively correlated with age at first primary and the relative risk remained substantially constant over time.


Transbronchial lung biopsy through the fibreoptic bronchoscope. Fifty-five patients were investigated using transbronchial lung biopsy through the flexible fibreoptic bronchoscope under fluoroscopic guidance, 51 by the transnasal route under local anaesthesia, two via an endotracheal tube whilst undergoing assisted ventilation and two by the same route under general anaesthesia. Multiple specimens of lung tissue subjects with diffuse pulmonary disease and in 16 of 32 (50%) with localized lesions, giving a positive yield of 64% of all cases. The only complication was an haemoptysis of 50 ml in one patient. Our results suggest that transbronchial lung biopsy is a relatively safe procedure which gives results comparable with other lung biopsy techniques.


An evaluation of the tuberculosis agglutination test. The tuberculosis agglutination titre as described by Nicholls (1975) was measured without prior knowledge of the diagnosis in 27 patients with open pulmonary tuberculosis, 100 patients with other diseases and 14 control patients with no apparent disease. Reproducibility of the test, as judged by repeat estimations on split samples, was poor. When attempting to distinguish between the disease categories the test gave 29% false negatives in patients with tuberculosis and 52% false positives in patients with other disease. Therefore the tuberculosis agglutination titre cannot be recommended for use in distinguishing tuberculosis from other conditions.


Sarcoidosis and selective IgA deficiency. Two patients are described in whom selective deficiency of IgA coexisted with a clinical condition compatible with a diagnosis of sarcoidosis. The literature on the rare association of the two conditions is reviewed. Any such association is probably coincidental.


Pleuropulmonary amoebiasis in Ghana: a report of three cases. Three cases of pleuropulmonary amoebiasis are reported for the first time from Ghana. It is suggested that where facilities for identification of the causal agent are not available a therapeutic trial should be given with drugs effective against Entamoeba histolytica.


Pulmonary alveolar microlithiasis.

Pulmonary alveolar microlithiasis. Three cases of pulmonary alveolar microlithiasis are reported at the symptom-free stage of this condition. Two were siblings.


Intrapleural haemorrhage complicating pulmonary arteriovenous fistula. Rupture of a pulmonary arteriovenous fistula caused intrapleural haemorrhage in a young woman with hereditary haemorrhagic telangiectasia and multiple bilateral fistulas. Bleeding was limited and exsanguination was probably prevented by pleural adhesions. Treatment included decortication, excision of the bleeding fistula, and prophylactic sclerosis of the opposite pleural space.


The pulmonary vasculature in haematite lung. A histological study was made of the pulmonary vasculature in two cases of haematite lung. Between fibrotic nodules in the lung the muscular pulmonary arteries showed the development of longitudinal muscle in the intima, a change thought to be associated with stretch and distortion of these vessels. In some arteries this muscular layer in the intima showed secondary fibrosis. No muscularized pulmonary arterioles were seen, indicating an absence of constriction of the terminal portions of the pulmonary arterial tree. Pulmonary arteries in fibrotic areas had become progressively engulfed by the fibrous tissue. The occlusive and obliterative vascular changes in 'haematite lung' are in reality those of silicosis which are brought about by the inhalation of silica particles with the iron sesquioxide. The only feature peculiar to haematite lung is the intense accumulation of iron-containing dust in and around the pulmonary blood vessels.