Detergent inactivation of sodium- and potassium-activated adenosinetriphosphatase of the electric eel. The stability of the sodium- and potassium-activated adenosinetriphosphatase (Na,K-ATPase) of the electric eel, Electrophorus electricus, was studied in five detergents in an effort to establish conditions for reconstitution of this membrane protein into defined phospholipids. The Na,K-ATPase activity of purified electric organ membranes as well as the ATPase is stable for at least 1 month of storage at 0 degrees C in the absence of detergents. At low concentrations of detergents, the enzyme is also stable for several days, but irreversible inactivation occurs rapidly as the detergent concentration is further increased. This inactivation begins at well-defined threshold concentrations for each detergent, and these concentrations generally occur in the order of the detergent critical micelle concentrations. Increasing the concentration of the electric organ membranes causes a linear increase in the inactivation threshold concentrations of Lubrol WX, deoxycholate, and cholate. The onset of inactivation evidently occurs when the mole fraction of detergent associated with the membrane lipids reaches a critical value in the narrow range of 0.2-0.4, in contrast to the large differences in the bulk concentrations of these detergents. The eel Na,K-ATPase is more sensitive to detergents than the sheep kidney enzyme.


Density distribution, characterization, and comparative aspects of the major serum lipoproteins in the common marmoset (Callithrix jacchus), a New World primate with potential use in lipoprotein research. Qualitative, quantitative, and comparative aspects of the serum lipoprotein profile in the Common marmoset (Callithrix jacchus), a New World primate, are described. Density gradient ultracentrifugation was used to evaluate lipoprotein distribution and to establish criteria for isolation of discrete molecular fractions. The major lipoprotein classes banded isopycnically on the gradient with the following hydrated densities: VLDL, d less than 1.017 g/mL; LDL, d = 1.027--1.055 g/mL; HDL fraction I, d = 1.070--1.127 g/mL; and HDL fraction II, d = 1.127--1.156 g/mL. Electrophoretic, immunological, and electron microscopic analyses attested to the purity of these fractions: the characteristics of each were assessed by chemical analysis, electron microscopy, immunological techniques, and polyacrylamide gel electrophoresis of their protein moieties. Marmoset VLDL and LDL were closely akin to those of man in size and chemical composition, although the former were richer in triglyceride; electrophoretic and immunological data showed the major protein component of VLDL and LDL to be a counterpart to human apo-B. The two HDL subfractions, i.e., HDL-I and HDL-II, corresponded in size and chemical composition to human HDL2 and HDL3, respectively, although slight differences in neutral lipid content were detected. By immunological and electrophoretic criteria, the major apolipoprotein of marmoset HDL was analogous to human apo-AI. In contrast, marked dissimilarities were evident in the complements of low molecular weight, tetramethylurea-soluble polypeptides of marmoset and human lipoproteins. Quantitatively, the human and marmoset lipoprotein profiles were not dissimilar, although HDL was the major class (approximately 50%); in fasting animals, serum concentrations of VLDL, LDL, and HDL were 50--90, 170--280, and 338--408 mg/dL, respectively. C. jacchus was distinct from man in displaying a greater proportion of its total HDL in the less dense (HDL-II) subfraction (marmoset HDL-I/HDL-II = approximately 4:1; human HDL2/HDL3 = approximately 1:3). These data indicate that, as an experimental animal for lipoprotein research, the Common marmoset combines the advantages of ready availability and maintenance with a serum lipoprotein profile which resembles, in many qualitative and quantitative aspects, that found in man.


Partial purification and characterization of a ribonucleic acid N2-guanine methyltransferase associated with avian myeloblastosis virus. A nucleic acid methylase, N2-guanine ribonucleic acid (RNA) methyltransferase, which is associated with type C RNA tumor viruses, has been purified from avian myeloblastosis virions by gel filtration on Sephadex G-200, followed by chromatography on hydroxylapatite. The molecular weight estimated by gel filtration is 220 000, and the methylase activity has a pH optimum of 7.6--7.9. Magnesium and ammonium ions both stimulate activity 1.5-fold at 9.5 mM and 0.36 M, respectively, but apparently neither is essential for activity. Both daunomycin and adriamycin, antineoplastic drugs, also increase activity 1.5-fold at 1 mM. The enzyme was purified 120-fold from the virions and the activity is partially stabilized by dithiothretiol, but large losses were sustained during 24-h dialysis. The purified enzyme retains 75% of its activity on storage at -25 degrees C for 2 months in buffer containing 50% glycerol. Escherichia coli tRNAPhe and tRNAVal are preferred substrates with methylation occurring at position 10 of E. coli tRNAPhe.


Purification and properties of the Ascaris pyruvate dehydrogenase complex. The pyruvate dehyhdrogenase complex (pyruvate:lipoate oxidoreductase (decarboxylating and acceptor-acetylating), EC 1.2.4.1) has been isolated from Ascaris muscle mitochondria and purified to near homogeneity by differential centrifugation, (NH4)2SO4 fractionation and calcium phosphate gel-cellulose chromatography. It is similar in shape, size and physical characteristics to pyruvate dehydrogenase complexes isolated from mammalian sources. It has an absolute dependence on CoA, NAD+ and pyruvate for activity and is competitively inhibited by acetyl-CoA and NADH. However, much higher NADH/NAD+ ratios are necessary to inhibit activity, suggesting regulation by the more reduced state of the pyridine nucleotide pool in Ascaris mitochondria.


Inhibition of (Na+ + K+)-ATPase by ouabain: involvement of calcium and membrane proteins. Treatment of plasma membrane isolated from murine plasmocytoma MOPC 173 with an EDTA-containing buffer resulted in a 300-fold increase in sensitivity of (Na+ + K+)-stimulated Mg2+-ATPase to ouabain. This phenomenon was associated with the solubilization by EDTA of phospholipid free proteins (approx. 30 000-34 000 daltons) from the cytoplasmic face of the plasma membrane and with removal of about 90% of the membrane bound Ca2+. The recovery of the original resistance to ouabain required specifically Ca2+ and was associated with a binding of the solubilized proteins to the membrane.


Contribution of villus and intervillus epithelium to intestinal transmural potential difference and response to theophylline and sugar. A chamber design is described which permits isolation of villus or intervillus epithelium from proximal segments of Amphiuma intestine and measurement of the transpithelial potential difference (psi ms) and short-circuit current (Isc) produced by each. In media containing Cl- and 10 mequiv./l HCO3- the villus generated a basal psi ms of 0.8 mV (serosa negative) and Isc of 12 microA/cm2 while the intervillus psi ms and Isc were not different from zero. Acetazolamide altered the villus psi ms by 1.2 mV; the intervillus psi ms by only 0.3 mV. Transepithelial gradients of HCO3- appeared to generate diffusion potentials across the intervillus but not the villus epithelium. The actively transported sugar galactose elevated psi ms by 0.6 +/- 0.1 mV in the intervillus epithelium and by 1.5 +/- 0.2 mV in the villus epithelium for a response ratio (0.6/1.5) = 0.4. The response ratio for valine was 0.3. In contrast, the response ratios for theophylline (0.7) and cyclic AMP (0.7) were significantly higher. These observations indicate that the entire epithelium is responsive to theophylline and cyclic AMP while Na+-dependent solute transport and the basal electrogenic ion transport processes are primarily functions of the cells lining the intestinal villus.


Inhibition of ouabain-binding to (Na+ + K+)ATPase by antibody against the catalytic subunit but not by antibody against the glycoprotein subunit. Antibodies against purified (Na+ + K+)ATPase from the rectal gland of Squalus acanthias, as well as against its catalytic subunit, inhibited ouabain binding by as much as 50%. However, antibodies against the glycoprotein subunit did not inhibit ouabain binding. These data suggest that binding of antibody against the catalytic subunit to the enzyme either covers the ouabain binding site or destroys its confirmation, while binding of antibody against the glycoprotein has no such effect.


Effects of dibutyryl cyclic AMP on the transport of alpha-methyl-D-glucoside and alpha-aminoisobutyric acid in separated tubules and brush border membranes from rabbit kidney. The effect of dibutyryl cyclic AMP on the transport of alpha-methyl-D-glucoside and alpha-aminoisobutyric acid in separated tubules and purified brush border membranes from rabbit kidney was investigated using a rapid filtration procedure. Dibutyryl cyclic AMP stimulated the uptake of alpha-methyl-D-glucoside and alpha-aminoisobutyric acid by separated renal tubules in agreement iwth prior studies utilizing renal slices (Rea, C. and Segal, S. (1973) Biochim. Biophys. Acta 311, 615--624; Weiss, I.W., Morgan, K. and Phang, J.M. (1972) J. Biol. Chem. 247, 760--764). However, in contrast to previous reports, no preincubation of the tissue with dibutyryl cyclic AMP was required for stimulation of transport to be manifest. Dibutyryl cyclic AMP stimulated oxygen consumption by separated tubules suggesting that stimulation of transport may occur by a linkage with renal oxidative metabolism. Dibutyryl cyclic AMP increased the uptake of alpha-aminoisobutyric acid into purified renal brush border membranes. However the uptakes of alpha-methyl-D-glucoside, proline, leucine and phosphate into brush border membranes were significantly inhibited.


A spin-label study of sciatic nerves from quaking, jimpy and trembler mice. The spin labels, 5-nitroxide stearic acid and 16-nitroxide stearic acid were incorporated into whole sciatic nerves dissected from normal, quaking, jimpy and trembler mice. With 5-nitroxide stearic acid, we have studied the thermal variation of the maximal apparent coupling constant (T) between 0 degrees C and 50 degrees C. Within this range of temperatures, we obtained identical values of 2 T for nerves from normal and jimpy mice, whereas 2 T was smaller for nerves from quaking and trembler mice. With 16-nitroxide stearic acid, composite spectra were recorded, particularly in the high-field range. A line characteristic of myelin was clearly observed in the spectra of nerves from normal and jimpy mice; its intensity was somewhat less in nerves from quaking mice and much less in spectra from trembler mice. A shoulder in the principal highfield line of the spectrum is modified only with nerves from jimpy mice. The results agree well with those obtained by electron microscopy, which reveal normal myelination in nerves from jimpy mice, a slight modification of the myelin from those of quaking mice and a practically complete demyelination in peripheral nerves from trembler mice. However, the structure of the nerves of jimpy mice also seems to be modified at an, as yet, undetermined level.


Effects of vanadate on ouabain binding and inhibition of (Na+ + K+)-ATPase. Effects of vanadate on ouabain binding and inhibition of sodium and potassium adenosine triphosphatase (Na+ + K+)-ATPase) were investigated under various ionic conditions. 1. Vanadate facilitated ouabain binding to (Na+ + K+)-ATPase in the presence of Mg2+ and this facilitation was partially reversed by catechol. 2. Vanadate antagonized the ability of high concentrations of NaCl to inhibit ouabain binding in the presence of magnesium. 3. Ouabain binding to the vanadate-enzyme complex, formed from magnesium and vanadate, was more sensitive to depression by potassium than that to the phosphoenzyme formed from magnesium and inorganic phosphate. 4. Preincubation of (Na+ + K+)-ATPase with vanadate in the presence of magnesium initially formed a potassium-insensitive complex as shown by a rapid initial rate of ouabain binding. However, within 5 min potassium overcame the vanadate potentiation of ouabain binding regardless of the order in which it was added to the reaction mixture. 5. Under conditions of enzyme turnover, vanadate failed to antagonize the inhibitory power of ouabain despite the presence of a high concentration of potassium. This suggests a possible relationship between the sensitivity of the sodium pump in various tissues to the cardiac glycosides and intracellular vanadate concentrations.


Purification and characterization of DNA-relaxing enzyme from Haemophilus gallinarium. A DNA-relaxing enzyme capable of concerted nicking and closing of DNA backbone bonds has been purified from Haemophilus gallinarum by two chromatographic steps and gel filtration. The enzyme efficiently catalyzes the removal of superhelical turns from a negatively twisted DNA and requires Mg2+ for this activity. Slight removal of superhelical turns from a positively twisted DNA generated by binding of ethidium bromide is found, but only at high enzyme concentrations. The DNA-relaxing activity is inhibited markedly with heat-denatured DNA, whereas native DNA and RNA have almost no affect on this activity.


Differential and selective inhibition of cellular and herpes simplex virus DNA synthesis by arabinofuranosyladenine. The influence of 9-beta-D-arabinofuranosyladenine (beta araAdo) and of its anomer 9-alpha-D-arabinofuranosyladenine (alpha araAdo) was studied in non-infected cells and cells infected with herpes simplex virus type 1 (HSV-1) and HSV type 2 (HSV-2). alpha AraAdo is a strong inhibitor of proliferation of non-infected cells. Multiplication of HSV-1 and HSV-2 is not affected at all by alpha araAdo, while their growth is strongly inhibited by beta araAdo. alpha AraAdo exerts no effect on the incorporation of dThd into HSV DNA, but blocks the incorporation into host cell DNA. Its anomer, beta araAdo, affects the incorporation rate of both the viral DNA system and the host cell DNA system (the latter one to a lesser extent). alpha AraAMP is incorporated into newly synthesized cellular DNA but not into HSV DNA. Enzymic studies relevant that alpha araATP has no effect on the HSV DNA polymerase system but a high inhibitory potency in the host cell DNA polymerase alpha system. The anomeric form, beta araATP, is a sensitive inhibitor of HSV DNA polymerase while the cellular DNA polymerases alpha and beta are more refractory.


Presence of protein at the termini of intracellular adenovirus type 5 DNA. Adenovirus type 5 contains linear double-stranded DNA with protein covalently attached to the ends of the molecules. The presence of protein at the termini of intracellular viral DNA in adenovirus type 5-infected cells was investigated at different stages during the replication process. The intracellular viral DNA was isolated from the nuclei by lysis in 4 M guanidine hydrochloride. Electrophoresis on agarose gels of HsuI restriction enzyme fragments and sucrose gradient centrifugation were used to detect protein on intracellular viral DNA. After uncoating parental DNA still contains protein attached to the termini of the viral genome. Replicating and mature progeny viral DNA can also be isolated in the form of DNA-protein complexes. These complexes exhibit the same properties as the DNA-protein complex isolated from purified virions. These results suggest that the protein at the termini of intracellular viral DNA is identical to the protein attached to the 5'-ends of the DNA extracted from virions and that it is possibly involved in the replication of viral DNA.


Inhibition of cytomegalovirus-stimulated human cell ornithine decarboxylase by alpha-difluoromethylornithine. 1. The relationship between synthesis of putrescine, human cytomegalovirus DNA synthesis, cell DNA synthesis, and human cytomegalovirus replication has been studied. 2. Stimulation of ornithine decarboxylase activity by shifting low serum-arrested whole human embryo cells to high serum medium is inhibited more than 99% by 2.5 mM DL-alpha-difluoromethylornithine. The addition of DL-alpha-difluoromethylornithine to human cells arrested in low serum and subsequently stimulated by the addition of fresh high serum-containing medium, causes a greater percent inhibition of ornithine decarboxylase activity than when the drug is added to growing human cells. 3. Increased ornithine decarboxylase activity produced by infection of low serum-arrested human cells was inhibited by 5.0 mM of DL-alpha-difluoromethylornithine. However, at a concentration of 5.0 mM, neither DL-alpha-methylornithine nor DL-alpha-difluoromethylornithine affected human cytomegalovirus growth or was toxic to these cells. These data suggest that the increased putrescine synthesis produced by infection is not required for virus replication. 4. The addition of 5.0 mM DL-alpha-difluoromethylornithine had no effect on human cytomegalovirus DNA synthesis or human cytomegalovirus-induced stimulation of cell DNA synthesis. However, 5.0 mM DL-alpha-difluoromethylornithine significantly reduced the stimulation of cell DNA synthesis caused by treatment with mock infecting fluid.


Synthesis of sugar-modified nucleoside 5'-triphosphates with partially purified nucleotide kinases from calf thymus. The partial purification of some nucleoside monophosphate kinases (ATP:nucleosidemonophosphate phosphotransferases, EC 2.7.4.4) from calf thymus by chromatography on Blue Sepharose to remove interfering phosphatase activity is described. Their specificities towards nucleoside monophosphates modified in the sugar are investigated. Pyrimidine nucleoside monophosphate kinase is not very much affected by such modifications, whereas GMP kinase does not tolerate such alteration. The effect on AMP kinase is intermediate.


Inhibitory effect of a lethal toxic fragment of staphylococcal alpha-toxin on cyclic AMP-dependent protein kinase activity. The effect of a lethal toxic fragment of staphylococcal alpha-toxin on the activity of adenosine 3',5'-monophosphate(cyclic AMP)-dependent protein kinase was examined. 1. The lethal toxic fragment produced a dose-dependent decrease in both the binding of cyclic AMP to the regulatory subunit and phosphorylation activity of cyclic AMP-dependent protein kinase obtained from rabbit skeletal muscles up to a plateau at a 50% inhibitory effect. The decrease in the activity of protein kinase observed with low doses of the lethal toxic fragment (0.1 microM) resulted from a competitive inhibition, probably by its interaction with the cyclic AMP-binding site in the regulatory subunit molecule. 2. The effects of a lethal toxic fragment and epinephrine on the cyclic AMP level and protein kinase activity were investigated in the perfused rabbit heart slices. The lethal toxic fragment attenuated the stimulation of cyclic AMP-dependent protein kinase activity ratio by epinephrine. 3. It is suggested that the specific action of a lethal toxic fragment on the cellular membrane enzymes may be attributable to the inhibition of the cyclic AMP-dependent protein kinase activity.


Properties of rat liver nuclear protein kinases. Two cyclic nucleotide-independent protein kinases (ATP:protein phosphotransferase, EC 2.7.1.37) have been purified to homogeneity from rat liver nuclei. While these enzymes have many similar catalytic properties (preference for acid rather than basic proteins), they differ in molecular weight and subunit composition. Protein kinase NII will utilize ATP and GTP as phosphate donors while protein kinase NI will only effectively use ATP. Both enzymes reveal an unusual activation by Fe2+.


Dephosphorylation of purine mononucleotides by alkaline phosphatases. Substrate specificity and inhibition patterns. Three purine mononucleotides, adenosine-, inosine- and guanosine monophosphate, were used as substrates at pH 7.4 and at 10.4 for three alkaline phosphatases (orthophosphoric-monoester phosphohydrolase (acid optimum), EC 3.1.3.1) containing similar phosphate-binding serine groups at their esteratic sites. Substrate specificity was found for the enzymes from calf intestine and bovine liver. Alkaline phosphatase from Escherichia coli was nonspecific. A substrate-dependent and pronounced inhibition with the purine analogue 1,3-dimethyl xanthine was found for the enzymes from intestine and liver, but not for alkaline phosphatase from E. coli. A substrate-independent and pronounced inhibition was found for all three enzymes with the phosphomonoester p-nitrophenol phosphate as the inhibitor. Alkaline phosphatases may play an important role in the regulation of the intracellular content of purine mononucleotides.


Isolation and identification of a small molecular weight inhibitor of cyclic nucleotide phosphodiesterase from bovine brain. In a search for endogenous regulators for cyclic nucleotide phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17), we found that the ultrafiltrate of bovine brain homogenate contained a cyclic nucleotide phosphodiesterase inhibitor. The inhibitor-containing fraction was further purified by ion-exchange column chromatography and gel filtration chromatography. The purified inhibitor was found to be a small molecular weight compound which had a maximum absorption at 248 nm. This compound was identified by thin-layer chromatography and high-pressure liquid chromatography as hypoxanthine. We suggest that hypoxanthine may serve as an endogenous regulator for the hydrolysis of cyclic nucleotide by cyclic nucleotide phosphodiesterase.


Activation of cyclic AMP-dependent protein kinases in human gut adenocarcinoma (HT 29) cells in culture. Vasoactive intestinal peptide, secretin, catecholamines and prostaglandin E1 stimulate the accumulation of cyclic AMP in HT 29 cells (see Laburthe, M. et al. (1978) Proc. Natl. Acad. Sci. U.S. 75, 2772-2775). In the present work maximal activation of protein kinases has been obtained at similar or even lower concentrations of the effectors. Maximal stimulation also requires a phosphodiesterase inhibitor. Type I and type II cyclic AMP-dependent protein kinases from basal and stimulated cells have been characterized by DEAE-Sepharose chromatography. Further identidication of the kinase has been carried out by gel electrophoresis and assay of the enzymes in the gel slabs. Comparison of the radioautography patterns of high speed supernatant lysate from basal and stimulated cells shows: First, that one type I and two type II cyclic AMP-dependent protein kinases plus one or two major and two minor cyclic AMP-independent protein kinases are present in HT 29 cells. Second, that all three holoenzymes are fully dissociated upon maximal stimulation, while the activity of the independent kinases appears unchanged.


Iron binding proteins and influx of iron across the duodenal brush border. Evidence for specific lactotransferrin receptors in the human intestine. The ability of a range of homologous transferrin-like proteins to donate iron to pieces of human duodenal mucosa, was examined with an in vitro incubation technique. In contrast to serum transferrin and ovotransferrin, only lactotransferrin was able to yield its iron to intestinal tissue, but in an autologous system this protein was unable to donate iron to human reticulocyte preparations. Studies with 125I-labelled lactotransferrin and lactotransferrin dual-labelled with 59Fe and 125I, indicated that the intact protein is excluded from entry into the enterocytes. The experiments suggest that iron may be transported across the brush border after delivery to specific protein binding sites at the cell surface.


Influence of plasma calcium and vitamin D on bone collagen. Effects on lysine hydroxylation and crosslink formation. In chick bone collagen the degree of lysine hydroxylation and the magnitude of the crosslink ratio dihydroxylysinonorleucine/hydroxylysinonorleucine were both found to be inversely related to the concentration of plasma calcium. Lysine hydroxylation was also affected by a second factor related to vitamin D status.


Increased number of beta-adrenergic receptors in the hypertrophied myocardium. Development of cardiac hypertrophy is associated with depletion of endogenous catecholamine stores and increased inotropic response to exogenous catecholamines. A biochemical basis for these changes is provided by the observation that the number of cardiac beta-adrenergic receptors - as reflected in specific [3H]dihydroalprenolol binding - is increased in hypertrophy without a change in the affinity of dihydroalprenolol for the binding sites or in the capacity of isoproterenol to displace dihydroalprenolol. This change in beta-receptor numbers may be an important adaptive mechanism for preserving the contractile performance of the hypertrophied myocardium.


Kinetic control of phosphoglycerate mutase from Flavobacterium sp. grown on ethylene glycol. A species of Flavobacterium able to oxidise ethylene glycol to pyruvate via glyoxylate, glycerate, 2-phosphoglycerate and phosphoenolpyruvate exploits phosphoglycerate mutase to initiate gluconeogenesis. Partially purified phosphoglycerate mutase from this bacterium is independent of adenylate charge control but is activated by phosphoenolpyruvate. The possible significance of this regulation is discussed.


The nature of the hepatic receptors involved in vasopressin-induced glycogenolysis. We have found a close correlation between the known vasopressor potency of arginine vasopressin and fourteen structural analogs, and the ability of these peptides to activate glycogen phosphorylase in isolated rat hepatocytes; there was no relation with the known antidiuretic activity of the analogs. We have also found that the pA2 values characterizing the known antivasopressor capacity of five analogs against vasopressin were close to those obtained for their inhibition of the vasopressin-induced activation of hepatic glycogen phosphorylase. We propose therefore that the hepatic receptors responsible for the glycogenolytic activity of vasopressin share characteristics with and appear therefore related to those responsible for pressor activity in vivo.


The protein inhibitor of adenosine 3':5'-monophosphate-dependent protein kinases. Isolation and characterization of three isoinhibitors. The protein inhibitor of adenosine 3':5 monophosphate-dependent protein-kinases has been purified from rabbit skeletal muscle by affinity chromatography on Sepharose-bound catalytic subunit of the kinase (Demaille et al. (1977) Biochemistry 16, 3080-3086). The inhibitory material can be separated into three species A, B and C either by polyacrylamide gel electrophoresis or by anion-exchange on DEAE-cellulose. However, the isoinhibitors display the same molecular weight and isoelectric point, and the same absence of acid-stable covalently bound phosphate. Their amino acid compositions are strikingly similar and their NH2-terminus are blocked. Also, their COOH-terminus appear to be very close to one another when studied by carboxypeptidase Y digestion. Their major difference lies in their inhibitory properties which are significantly weaker in inhibitor C (Ki = 0.87 nM) than in A and B (Ki = 0.18 and 0.23 nM, respectively). This is the first report on the existence of various forms of the protein-kinase inhibitor that exhibit different inhibitory properties and may play a role in the regulation of the protein-kinase system.


Oxidation-reduction potentials of the hemes in cytochrome C3 from Desulfovibrio gigas in the presence and absence of ferredoxin by EPR spectroscopy. 1. Ferricytochrome c3 from D. gigas exhibits two low-spin ferric heme EPR resonances with gz-values at 2.959 and 2.853. Ferrocytochrome c3 is diamagnetic based on the absence of any EPR signals. 2. EPR potentiometric titrations result in the resolution of the two low-spin ferric heme resonances into two additional heme components representing in total the four hemes of the cytochrome, with EM values of -235 mV and -315 mV at heme resonance I and EM values of -235 mV and -306 mV at heme resonance II. 3. EPR spectroscopy has detected a significant diminution of intensity (approx. 60 p. 100) in the gx amplitude of ferricytochrome c3 in the presence of D. gigas ferredoxin II. The presence of ferredoxin II also causes a more negative shift in the EM of the second components of the signals at heme resonances I and II of cytochrome C3. Both observations suggest that an interaction has occurred between cytochrome C3 and ferredoxin II. 4. The results presented suggest that the heme ligand environment of ferricytochrome c3 from D. gigas is less perturbed and/or less asymmetric than environment for ferricytochrome c3 from D. vulgaris whose EPR behavior indicates the non-equivalence of all four hemes.


Chromatography of a Triton extracted beef cardiac cytochrome C1 preparation on diethylaminoethyl cellulose. During the chromatography of a Triton X-100 extracted preparation of the mitochondrial membrane proteins on diethylaminoethyl cellulose, we have observed two chromatographic fractions containing cytochrome c1. One elutes from diethylaminoethyl cellulose with aqueous buffers alone, and the other elutes with those buffers after the addition of the nonionic detergent Triton X-100. The two forms occur in equimolar ratios and each retains its original chromatographic character on rechromatography, with no conversion of one form into the other.


Pattern generation in the lobster (Panulirus) stomatogastric ganglion. II. Pyloric network simulation. 1. Results from the companion paper were incorporated into a physiologically realistic computer model of the three principal cell types (PD/AB, LP, PY) of the pyloric network in the stomatogastric ganglion. Parameters for the model were mostly calculated (sometimes estimated) from experimental data rather than fitting the model to observed output patterns. 2. The initial run was successful in predicting several features of the pyloric pattern: the observed gap between PD and LP bursts, the appropriate sequence of the activity periods (PD, LP, PY), and a substantial PY burst not properly simulated by an earlier model. 3. The major discrepancy between model and observed patterns was the too-early occurrence of the PY burst, which resulted in a much shortened LP burst. Motivated by this discrepancy, additional investigations were made of PY properties. A hyperpolarization-enabled depolarization-activated hyperpolarizing conductance change was discovered which may make an important contribution to the late phase of PY activity in the normal burst cycle. Addition of this effect to the model brought its predictions more in line with observed patterns. 4. Other discrepancies between model and observation were instructive and are discussed. The findings force a substantial revision in previously held ideas on pattern production in the pyloric system. More weight must be given to functional properties of individual neurons and less to properties arising purely from network interactions. This shift in emphasis may be necessary in more complicated systems as well. 5. An example has been provided of the value quantitative modeling can be to network physiology. Only through rigorous quantitative testing can qualitative theories of how the nervous system operates be substantiated.


[Electron carriers of adrenocortical mitochondria. Terminal systems]. A procedure for isolation of cytochrome oxidase and cytochrome P-450 from adrenocortical mitochondria was developed. The heme and copper contents, subunit composition, optical and EPR spectra for these enzymes were determined. The effects of pH, substrates and some inhibitors on the spectra of cytochrome P-450 were studied. It was found that cytochrome oxidase did not inhibit the reactions catalyzed by cytochrome P-450; cytochrome P-450 had no inhibiting effect on the oytochrome oxidase activity.


[Electron acceptors in photosynthetic reaction centers from Rhodopseudomonas spheroides]. Using optical differential spectroscopy and EPR, a parallel study of light-induced electron transfer between the primary (X1) and secondary (X2) quinone-like acceptors in the preparations of reaction centers (RC) isolated from bacterial chromatophore membranes with sodium dodecyl sulfate was carried out. The data from direct measurements of the rate constant temperature dependence for the interaction between light-reduced X1 and X2 (KX1X2) are in good agreement with the data calculated from the kinetic analysis of dark reduction of photooxidized bacteriochlorophyll RC on the acceptors X1 and X2 (KX1X2 = 2.10(-1)S at 20 degrees; Ea = 11,8 kcal.mol-1 within the temperature range of 20 degrees-- -20 degrees). This evidence proves the efficiency of the previously used approach /1, 2/ for the evaluation of the X1-X2 interaction. The method proposed was used for a kinetic analysis of a low-temperature electron transfer from X1 to X2 in RC isolated with lauryldimethylaminoxide (KX1X2 = 2,3.10(2) S-1 at 20 degrees; Ea = 5,5 kcal.mol-1 within the temperature range of 10 degrees-- --70 degrees).


[Metlegoglobin reductase from lupine nodules. Purification and properties]. The purification procedure and properties of metlegoglobin reductase from the soluble fraction of lupine (Lupinus luteus L.) nodules and from the proteins secreted by bacteroids Rhizobium lupini in vitro are described. The properties of both forms of enzyme were found to be similar. A metlegoglobin reductase preparation purified 125-fold with a yield of 21% was obtained. The enzyme is strictly specific to the cofactor (NADH). No substrate specificity was revealed. The enzyme reduces oxidized cytochrome c, Mb+, Lb+, Hb+ and exygen. The pH optimum for the enzyme is 7,4. The enzyme is inhibited by p-chloromercurybenzoate. In some properties the enzyme from lupine nodules is close to methemoglobin reductase from the erythrocytes. It was shown that apart from metlegoglobin reductase, bacteroids secrete some other proteins, which is indicative of a close interrelationship between the bacteroids and the plant in a symbiotic nitrogen-fixing system.


[Metabolic characteristics of prolyl hydroxylase]. The turnover time of prolyl hydroxylase in the skin of one-day-old rats treated in vivo with L-[14C]leucine is 22--34 hrs. Theoretically this time interval comprises two processes: i. e. incorporation of [14C]-Leu into the inactive enzyme precursor and transformation of the inactive from into the active enzyme.


[Possible role of high molecular weight polyphosphates in ATP synthesis from exogenous adenine by the culture of Corynebacterium sp., strain VSTI-301]. An addition of exogenous adenine to an autolysing 72-hour culture of Corynebacterium sp., strain BSTI-301 results in accumulation of as much as 0,6--1,0 mp of ATP per 1 ml of medium. Extracellular ATP accumulation under such conditions is coupled with a considerable decrease of the intracellular content of 5'-phosphoribosyl-1-pyrophosphate, orthophosphate, pyrophosphate and two fractions of high-polymeric polyphosphates PPh3 and PPh4, as compared to the control. The activity of pyrophosphate phosphohydrolase (EC 3.6.1.1) and polyphosphate phosphohydrolase (EC 3.6.1.11) is thereby considerably decreased in the cells growing on exogenous adenine, while the activity of ADP-phosphotransferase (EC 2.7.4.1) is increased 2-fold. It was found that in experiments with 14C-adenine the intracellular content of both ATP and ADP remains unchanged despite a considerable accumulation of extracellular ATP in Corynebacterium sp., strain BSTI-301 cells.


[Secondary structure and methylation of DNA in reinoculated tumors and cultures of embryonic cells transformed by oncogenic viruses]. Using the kinetic formaldehyde method the concentration of secondary structure defects (SSD) in the DNAs of ascite leukosis L 1210 cells and cultures of hamster embryonic cells transformed by virus SV-40 and polyoma was evaluated. It was found that this concentration was considerably higher than in the DNAs from normal liver and primary culture of mouse embryonic cells. The occurrence of the defects in malignant cell DNAs is not due to enzymatic degradation of DNA. Using thin-layer chromatography the content of m5C in the DNAs from 17 sources (transformed cell cultures, experimental tumours and liver cells of mouses with Ehrlich ascite carcinoma) were determined. The methylation level for all these DNAs was higher than for normal mouse and rat liver DNAs. No correlation between the SSD concentration and m5C content in the DNAs studied was observed.


[Oxidation of 4-anilino-5-methoxydioxybenzene-1,2 in complex biochemical systems possessing superoxide dismutase activity]. The kinetics of 4-anilino-5-methoxydioxybenzene-1,2 (AMOBQH2) autoxidation in biochemical systems possessing the superoxide dismutase activity were studied. The autoxidation of AMOBQH2 is affected by superoxide dismutase, which is indicative of participation of the superoxide radical in this process. The main kinetic effect of superoxide dismutase consists in a decrease of the effective rate constant for AMOBQH2 autoxidation. Peroxidase releases the superoxide dismutase inhibition of AMOBQH2 autoxidation. The data obtained are discussed in terms of a biochemical mechanism of action of biologically active aminoaromatic derivatives of o-benzoquinone.


An automated analysis of REM sleep in primary depression. The REM sleep of 23 nonpsychotic patients with primary depression was studied by means of an automated REM analyzer during a drug-free period and again during amitriptyline administration. Initial drug administration (50 mg) was associated with an immediate reduction in the number, average frequency, and average size of the rapid eye movements. The average REM size remained suppressed with continued drug administration while the average REM frequency showed a rebound which was responsible for a partial recovery of the number of REMs and total REM intensity to predrug levels. With regard to individual REM periods, REM frequency and REM intensity were redistributed during tricyclic administration so that the second REM period became more "intense" than the first REM period. This automated REM analysis technique provides an objective set of measures for characterizing discrete aspects of REM sleep during a depressive episode and for evaluating the changes in REM sleep during psychotropic trials.


Narcolepsy: a family study.

Narcolepsy: a family study. We obtained medical and psychological assessments and 48-hr polysomnographic recordings on five sisters, three of whom had narcolepsy. Of the three, two were identical twins. All three narcoleptic sisters cited emotional stress and environmental demands for sustained performance as the major factors which aggravated their symptoms, and corresponding to this, the illness followed a different life course in each of the three. Most striking were the differences between the twin sisters in clinical symptoms and polysomnographic signs. One sister suffered from all the symptoms of narcolepsy and her sleep recording showed the typical sleep onset REM periods of the disease. Her twin suffered only from excessive daytime drowsiness and her sleep recording was normal--at least by the usual criteria. The sleep of all three narcoleptic sisters, however, was significantly more fragmented than that of their normal siblings. Our data suggested that excessive sleep fragmentation was a basic feature of narcolepsy and that it betrayed a constitutional predisposition for sleep to dissociate into its components and to become distributed around the nycthemeron. This process could be aggravated by emotional stress and by environmental demands for sustained vigilance, and this in turn, created the differences in symptoms and signs between individuals with identical genetic predispositions.


Preparation and properties of immobilized flavokinase. Flavokinase (ATP: riboflavin 5'-phosphotransferase, EC 2.7.1.26) purified from rat liver by affinity chromatography, has been immobilized by amide linkage to omega-aminoalkyl-agarose beads. The immobilized enzyme differs from the soluble enzyme in having greater stability, slightly higher Km for the substrates, riboflavin and ATP, a broader pH optimum, and a lower energy of activation. These results suggest that the immobilized enzyme is influenced by the microenvironment of the bead and is subject to some degree of internal diffusional limitation. A small (3 ml), continuous, plug-flow reactor prepared with immobilized flavokinase effects 50% conversion of riboflavin to riboflavin 5'-phosphate (FMN) with a flow rate of 0.16 ml/min, which corresponds to an output of 5 nmol FMN/min. Immobilized flavokinase is effective for phosphorylating riboflavin and numerous riboflavin analogs and provides a facile method for preparing exclusively, unlike other synthetic methods, the 5'-phosphates.


[Reaction of nonmyelinated fibers of the cutaneous nerve of cats to cooling]. Combining the colliding impulses method with the methods of singling out the nerve's weak signals from the equipment noise shows that most nonmyelinated fibers of C1 and C2 groups develop impulses under cooling the cat hairy skin. In some nonmyelinated fibers of the C2 group there takes place an inhibition of spontaneous impulses in response to cooling.


[Effect of Vibrio cholerae filtrate on the adenyl cyclase system in vitro]. An in vitro model using homogenates of the rat intestine and liver for studying the V. Cholerae culture filtrate effect on the adenylate cyclase system is proposed. Optimal conditions for the adenylate cyclase functioning have been investigated for this model. It was shown that V. Cholerae filtrate induces a stable activation of adenylate cyclase and does not change the activity of cyclic AMP phosphodiesterase. It was also established that the activity of phosphodiesterase in the intestinal homogenate is about 2.5 fold higher than that in the liver homogenate. The model may be considered promising for investigation of the mechanism of cholerae toxin action.


[Thymic Kurloff inclusion. Action on the ovogenesis of the rat]. Grafts and injections of cellular extracts from thymus and spleen of guinea pigs, were carried out in puberal and impuberal female rats. These organs are especially rich in Foa-Kurloff cells after oestrogentherapy. Kidney served as a reference material for the same experimental conditions. Controls received cellular injections and grafts from guinea pig organs without treatment. In all cases, ovarian histological and biometric study of recipient animals shows no alteration of the ovarian cycle. On the other hand, grafts and injections of cellular extracts from organs with high concentration of Foa-Kurloff cells induced hyperactivity of internal theca folliculi and of ovarian interstitial cells, only in puberal female rats.


Radiology of the adult sella turcica.

Radiology of the adult sella turcica. Radiographic changes of the sella turcica or pituitary fossa reflect multiple disease processes of intracranial, cranial and systemic origin. Their value as diagnostic aids is related to the fact that the development of the fossa depends somewhat upon that of the pituitary gland, which it partially encloses. Further, many vascular and neural structures about the base of the brain lie close to the fossa. Also, it is vulnerable to increased intracranial pressure. The ease of radiographic demonstration of the sellar components from many planes and angles allows early recognition of pathologic alterations. Radiographic anatomy, both normal and abnormal, will be discussed, as well special diagnostic procedures such as computed tomography, angiography and cerebral pneumography.


Heavy particle therapy: pituitary tumors. Therapy with 910 MeV alpha particles provides a treatment with no mortality and an extremely low morbidity and has been highly successful in the control of pituitary hormone hypersecretion and tumor growth. It is possible to deliver radiation doses to the pituitary gland that are sufficiently high to inhibit or destroy the tumor cells that cause abnormal secretory activity without damaging the surrounding structures. The data demonstrate the effectiveness of this form of treatment in acromegaly, non-functioning pituitary tumors, prolactin secreting adenomas, Cushing's disease and Nelson's syndrome.


Surgery of the pituitary gland.

Surgery of the pituitary gland. Increasing diagnostic sophistication among endocrinologists has continually expanded the horizon of surgical treatment of adenohypophyseal problems. This advancement in endocrinology has been paralleled by improvement in the technical sophistication of neurosurgeons which has resulted in an increasingly aggressive approach to these serious disorders. Neurosurgical treatment is clearly able to provide resolution of significant physical disorders with minimal sequelae. The apparent potential for transsphenoidal approaches to the sella turcica as described within this paper has intrigued a number of neurosurgeons throughout this century. Their continuing interest has carried this procedure to a secure position in the management of many of these complex problems. This paper describes the author's experience with 132 consecutive transsphenoidal hypophysectomies.


Clinical pituitary disorders.

Clinical pituitary disorders. The pituitary gland is really a confederation of several relatively independent endocrine glands gathered together in close relationship to the hypothalamus which exerts considerable governing control. Each of these sub-glands of the pituitary should be considered separately in every case of pituitary disease. While such an approach is mandatory in dealing with laboratory investigation, it also is profitable in considering clinical and therapeutic aspects of each case to ensure that nothing is overlooked.


Pathology of pituitary adenomas.

Pathology of pituitary adenomas. A new pituitary classification has been developed, based on the histologic, histochemical, immunocytologic and electron microscopic investigation of 207 pituitary adenomas removed surgically from male and female patients of various ages and different clinical symptomatology. Based principally on cytogenesis, the classification attempts to correlate the morphologic features of the tumor cells with their secretory activities, the clinical history, symptomatology, and biochemical findings. The classification consists of the following eight entities: 1) growth hormone cell adenoma; 2) prolactin cell adenoma; 3) mixed growth hormone cell-prolactin cell adenoma; 4) acidophil stem cell adenoma; 5) corticotroph cell adenoma; 6) thyrotroph cell adenoma; 7) gonadotroph cell adenoma; 8) undifferentiated cell adenoma, including oncocytoma. Prolactin adenomas were found to be the most frequently occurring pituitary adenoma type.


Investigation of the role of extracellular calcium in the control of acid secretion in the isolated whole stomach of the rat. 1 An isolated stomach preparation from immature rats has been used to study the role of extracellular calcium in the control of gastric acid secretion. Calcium was removed from both the serosal and mucosal solutions either in the absence of a chelating agent or in the presence of EGTA.2 Removal of calcium in the absence of EGTA had no significant effect on basal acid secretion or the acid responses to gastrin and dibutyryl cyclic adenosine 3',5'-monophosphate (db cyclic AMP). Under the same conditions there was a marked potentiation of the acid response to histamine, and a reduction of the acid response to acetylcholine which was readily reversed on restoring calcium to the bathing solutions.3 Removal of calcium in the presence of EGTA caused an inhibition of basal acid secretion and of the acid responses to histamine and db cyclic AMP. In each case this reduction in acid output was readily reversed on bathing the stomachs in normal calcium-containing (2.5 mM Ca(2+)) EGTA-free solutions.4 The inhibition of the acid response to histamine produced by Ca(2+)-free solutions which contained EGTA was not reversed on bathing the stomachs in solutions that contained both EGTA (0.1 mM) and an excess of calcium (2.5 mM).5 The removal of extracellular calcium in the absence of EGTA provided evidence that the secretion of H(+) ions is dependent under some conditions on calcium ions. The possibility cannot be excluded that EGTA itself exerts an inhibitory influence on the process of acid secretion.


The effects of calcium and magnesium ions, temperature and repetitive stimulation on inhibitory junctional transmission in smooth muscle of guinea-pig small intestine. 1 The effects of calcium and magnesium ions and temperature on the peak amplitude of the nonadrenergic, noncholinergic inhibitory junction potential, evoked by a single stimulus, or paired transmural stimuli, were examined in the circular muscle of guinea-pig small intestine.2 The peak amplitude of the inhibitory junction potential (i.j.p.) could be decreased by lowering the external concentration of calcium or by raising the external magnesium concentration (at 25 degrees C).3 At 25 degrees C, the second of a pair of i.j.ps was larger than the first at short intervals (<0.2 s), but smaller at larger intervals (0.2 to 20 s). Enhancement of the second (test) response decayed exponentially with a time constant of 87 ms. Depression of the test i.j.p. was maximal at 0.4 s and then recovered exponentially with a time constant of 11 s.4 In low calcium or high magnesium solution, depression of the test i.j.p. decreased without any change in the rate of recovery from depression.5 Despite the slow rate of recovery from depression after a single conditioning response, transmitter output could be maintained during low-frequency repetitive stimulation.6 The peak amplitude of the i.j.p. increased as the temperature was raised to 35 degrees C (Q(10) = 1.5).7 In contrast to the neuromuscular junction, the depression of the second of a pair of i.j.ps decreased as the temperature was raised. At 35 degrees C the test i.j.p. was larger than the conditioning i.j.p. for most stimulus intervals (1 to 20 s).8 The results suggest that the rate of replenishment of the store of inhibitory transmitter is sensitive to both temperature and repetitive stimulation.


Naloxone reverses the antihypertensive effect of clonidine. In unanesthetized, spontaneously hypertensive rats the decrease in blood pressure and heart rate produced by intravenous clonidine, 5 to 20 micrograms/kg, was inhibited or reversed by nalozone, 0.2 to 2 mg/kg. The hypotensive effect of 100 mg/kg alpha-methyldopa was also partially reversed by naloxone. Naloxone alone did not affect either blood pressure or heart rate. In brain membranes from spontaneously hypertensive rats clonidine, 10(-8) to 10(-5) M, did not influence stereoselective binding of [3H]-naloxone (8 nM), and naloxone, 10(-8) to 10(-4) M, did not influence clonidine-suppressible binding of [3H]-dihydroergocryptine (1 nM). These findings indicate that in spontaneously hypertensive rats the effects of central alpha-adrenoceptor stimulation involve activation of opiate receptors. As naloxone and clonidine do not appear to interact with the same receptor site, the observed functional antagonism suggests the release of an endogenous opiate by clonidine or alpha-methyldopa and the possible role of the opiate in the central control of sympathetic tone.


Modulation of sympathetic transmission by neuronally-released dopamine. 1 When rabbits were pretreated with Fla-63, there was a marked inhibition of dopamine-beta-hydroxylase such that, after incubation of the ear arteries with [3H]-dopamine 47.2% of the tritium in the tissue was retained as unchanged dopamine. 2 [3H]-dopamine was released by stimulation of the sympathetic nerves in ear arteries taken from rabbits pretreated with Fla-63 and incubated with [3H]-dopamine. 3 The dopamine antagonists metoclopramide (1.0 microM) and ergometrine (1.0 microM) enhanced the stimulation-induced efflux of tritium in ear arteries taken from rabbits pretreated with Fla-63 and incubated with [3H]-dopamine, but not when the arteries were incubated with [3H]-noradrenaline. 4. These results suggest that if dopamine is present in the transmitter stores, it can be released by stimulation of the sympathetic nerves, and if the amount is adequate, it can activate an inhibitory feedback loop where prejunctional dopamine receptors are present.


Effects of the venom of the green mamba, Dendroaspis angusticeps on skeletal muscle and neuromuscular transmission. 1 The venom of the green mamba, Dendroaspis angusticeps, was tested for effects on neuromuscular transmission and skeletal muscle contractility in isolated phrenic nerve-hemidiaphragm preparations of the rat and mouse, chick biventer cervicis muscle preparations and in aneural cultures of embryonic chick skeletal muscle. 2 The venom (10 to 40 micrograms/ml) augmented the responses to indirect but not direct stimulation. As the venom did not have anticholinesterase activity and did not increase receptor sensitivity, it is likely that the venom enhanced release of acetylcholine. 3 Higher concentrations of venom (40 to 80 micrograms/ml) inhibited acetylcholine receptor sensitivity. 4 Prolonged exposure to the higher concentrations of venom produced a failure of muscle contractility. Signs of muscle degeneration were seen in skeletal muscle cultures.


The failure of indomethacin to alter ACTH-induced adrenal hyperaemia or steroidogenesis in the anaesthetized dog. 1 The response of adrenal blood flow to adrenocorticotropic hormone (ACTH) was measured with radioactive microspheres in anaesthetized, dexamethasone-treated, mongrel dog. 2 Adrenocorticotropic hormone (2 u/h i.v.) increased adrenal blood flow within 15 min and this persisted for the duration of the infusion. 3 Cortisol concentrations also rose with ACTH infusion. 4 Indomethacin (6 mg/kg i.v. followed by 1 mg/min) did not effect the adrenal response to ACTH although plasma concentrations of indomethacin (21.9 +/- 2.5 micrograms/ml) adequate to suppress prostaglandin synthesis were achieved. 5 We conclude that prostaglandins are not required for steroidogenesis or the adrenal haemodynamic response to ACTH.


The effects of inert gases and other general anaesthetics on the release of acetylcholine from the guinea-pig ileum. 1 The actions of a range of general anaesthetic agents on the rates of release of acetylcholine from the guinea-pig ileum were tested, by means of a superfusion system designed to maintain the tissues under physiological conditions in a high pressure chamber.2 Anaesthetic pressures of nitrous oxide, nitrogen, argon, sulphur hexafluoride and carbon tetrafluoride caused increases in acetylcholine ouput but the concentrations required did not parallel their general anaesthetic potencies. The changes were not altered by the application of a pressure of helium which reverses their general anaesthetic actions in vivo.3 Urethane (50.5 mM and 101 mM, but not 16.8 mM) decreased acetylcholine release rates and this effect was not reversed by helium pressure.4 Octanol (1.0 mM, but not 0.124 mM or 0.496 mM) decreased the acetylcholine output. This action was not reversed by helium pressure. The lack of effect on acetylcholine release from tetrodotoxin-treated tissues suggested that the changes were produced by blockade of action potential conduction.5 Phenobarbitone (0.4 mM but not 0.2 mM) also decreased acetylcholine output. Although the concentrations required were lower than those which have been previously shown to block axonal conduction, no changes were seen in tetrodotoxin-treated tissues. The decreases were less when helium pressure was applied than at atmospheric pressure but full pressure reversal, as occurs in vivo, was not seen.6 The effects on acetylcholine output exerted by the anaesthetics studied did not reflect their general anaesthetic action in the concentrations required, the direction of the changes produced or in the response to helium pressure. They represent specific actions which are likely to contribute to the individual differences which are seen between the physiological actions of the anaesthetics in vivo.


The subdivision of beta-adrenoceptors in the cardiovascular system of the rat. 1 The antagonism by the beta-adrenoceptor blocking drugs, propranolol (non-selective) and practolol (beta-selective), of the cardiovascular actions of isoprenaline has been investigated in the rat. 2 All doses of practolol (0.1, 1 and 3 mg/kg) blocked the cardio-accelerator action of isoprenaline but only the largest dose blocked the vasodilator effect. 3 All doses of propranolol (0.01, 0.03 and 0.1 mg/kg) blocked the vasodilator effect of isoprenaline but only the largest dose diminished the tachycardia. 4 It is concluded that in the rat, as in other species, beta-adrenoceptors may be subdivided into beta 1 (cardiac) and beta 2 (peripheral vascular) types.


Nephroblastoma: treatment during 1970-3 and the effect on survival of inclusion in the first MRC trial. In 1970-3 313 children were diagnosed as having nephroblastoma in Great Britain. From the start of the first Medical Research Council nephroblastoma study in October 1970 until the end of 1973, 98 children (57% of all eligible children) were included in the trial. Of the 313 children, 288 (92%) had a nephrectomy, 248 (79%) received a course of radiotherapy, and 267 (85%) were given at least four days' chemotherapy. The three-year survival rate was 58%; the rate among children in the trial (77%) was significantly better than that among children who were eligible for the trial but not included (58%). Children who had nephrectomies at specialised children's and teaching hospitals had a higher survival rate than those treated elsewhere. All children with nephroblastoma should be treated according to well-defined protocols which take into account the age of the child and the stage of the tumour and include a full course of maintenance chemotherapy.


Tegmentoreticular projections with special reference to the muscular atonia during paradoxical sleep in the cat: an HRP study. Using the retrograde tracer technique with horseradish peroxidase (HRP), attempts were made to determine the cells of origin and the descending pathway of the tegmentoreticular projections in order to give an anatomical substrate for the physiological phenomenon of the postural atonia observed during paradoxical sleep (PS) in the cat. The HRP was injected into various parts of the pontomedullary reticular formation (RF) including the caudal raphe nuclei, nucleus (n.) reticularis gigantocellularis (Gc), n. reticularis magnocellularis (Mc), and other pontomedullary structures adjacent to the Mc. The results indicated that the HRP injection into the Mc, particularly its caudal and lateral two-thirds, resulted in specific labeling of cells located in an area just medial to the LCa together with those in the most medial part of the LCa. Bilateral lesions of these pontine structures have been reported to suppress the atonia otherwise observed during PS in the normal cat. In addition to the HRP labeled cells, we have also observed HRP filled fiber bundles directed to labeled cells in the medial part of the LCa and immediately adjacent tegmental RF area. The same course of HRP labeled fiber bundles was also observed together with HRP labeled cells in the Mc after HRP injections into the medial part of the LCa area, indicating the existence of an interconnection between the LCa area and the Mc. The location of the tegmentoreticular pathway corresponded to that of the lesions effective to suppress the muscular atonia during PS. HRP injections into the caudal medullar caudal to the Mc, on the other hand, resulted in no or almost no HRP labeled cells in the area medial to the LCa, in spite of the presence of HRP containing neurons in other parts of the pontomedullary RF areas, showing that the tegmentoreticular projections as described above terminate almost exclusively in the Mc.


The depressive effects of morphine on the C fibre response of dorsal horn neurones in the spinal rat pretreated or not by pCPA. (1) The effects of morphine upon the transmission of nociceptive messages at the spinal level have been investigated in the spinal rat. The responses of dorsal horn cells induced by the activation of C fibres were depressed in all cases in a dose-dependent fashion, this effect being reversed by the opiate antagonist naloxone. An estimation of the ED50 at the cellular level leads to the value of 6.3 mg/kg. The responses to A delta fibres were also depressed dose-dependently whereas the responses to A alpha fibres were unaffected. This is a confirmation in the rat of the differential effects of morphine on responses of convergent units elicited by the stimulation of different fibres, as previously described in the cat. (2) The hypothesis of the participation of serotonergic terminals in these effects has been checked by comparing the preceding results to those obtained in pCPA pretreated animals. Two populations of units were observed in the latter group: two-thirds of cells showed a dose-response curve similar to that of the non-pretreated group whereas the remaining one-third were unaffected either by morphine or naloxone. It is concluded that, at least, two mechanisms are involved in the depressive effects of morphine at the spinal level, serotonergic terminals being implicated in one of these. (3) The lowering of spinal cord serotonin content was associated with a decrease of both the size of the excitatory receptive field (34%) and the activities related to C fibre input (36%) of the recorded dorsal horn cells. This result is discussed with reference to the excitatory or sensitizatory effect of serotonin upon chemoreceptors related to pain.


The influence of naloxone on the C fiber response of dorsal horn neurons and their inhibitory control by raphe magnus stimulation. In intact rats anesthetized with chloralose, the effects of naloxone were studied on the responses of spinal cord dorsal horn neurons to C fiber stimulation and upon the inhibition induced on these responses by stimulation of the nucleus raphé magnus (NRM). (1) A mean 44% facilitatory effect on responses to C fibers was observed for 12/19 units. (2) A mean 30% reduction of the inhibitory effects of NRM was found for 14/29 units. (3) However there is no clear relationship between these facilitatory effects and the diminution of the efficiency of NRM stimulation. These results demonstrate a facilitatory effect of naloxone upon the transmission of noxious messages at the spinal level and confirm that opiate endogenous substances are implicated in the inhibitory mechanisms activated by stimulation of NRM.


The localization of receptor binding sites in the substantia nigra and striatum of the rat. Neurotransmitter receptor binding of 5 ligands was examined in the striatum, substantia nigra (SN) and frontal cortex of rats which had received either unilateral 6-hydroxydopamine (6-OHDA) lesions of the nigrostriatal pathway (NSP) or unilateral kainic acid lesions of the striatum. 6-OHDA lesions of the NSP significantly reduced [3H]dihydroalprenolol ([3H]DHA) and [3H]naloxone ([3H]Nal) binding by 31% and 28% respectively, in the denervated striatum compared to the contralateral side. Scatchard analysis revealed that the alteration in [3H]DHA binding was not due to a change in the affinity of the beta-adrenergic receptor for [3H]DHA. In marked contrast to these changes in the striatum, destruction of the NSP resulted in a significant increase in [3H]DHA and [3H]Nal binding by 44% and 26%, respectively, in the frontal cortex of the lesioned compared to the control side. 6-OHDA lesions in the NSP did not alter striatal receptor binding for [3H]quinuclidinyl benzilate ([3H]QNB), [3H]muscimol ([3H]Mus) or [3H]flunitrazepam ([3H]Flu). Similarily, intrastriatal kainic acid injections did not alter striatal receptor binding for [3H]Nal, [3H]Flu or [3H]Mus. Of the various receptor densities measured in the SN after the above lesions the only alteration observed was a 43% increase in [3H]Flu binding following 6-OHDA lesions of the NSP. Scatchard analysis indicated no change in the affinity of the benzodiazepine receptor for [3H]Flu. 6-OHDA lesions of the NSP did not alter [13H]QNB or [3H]Nal binding in the SN. Striatal kainic acid lesions did not alter nigral [3H]QNB or [3H]Flu binding. The results are discussed in terms of neurotransmitter localization and plasticity within the striatum, SN and frontal cortex.


Biochemical and immunohistochemical studies on dysmyelination of quaking mutant mice in vivo and in vitro. Dysmyelination in the central nervous system of the quaking mutant mouse was studied biochemically and immunohistochemically. We found, by measuring CNPase activity, that myelination in the central nervous system of quaking mice was affected to a different degree in different areas. The pallium cerebri was the most severely affected and the medulla and spinal cord were least affected. The density of astroglia observed by GFA staining wash higher in the white matter of quaking mice than in controls, but the total area of the white matter in the cerebellum was smaller in the quaking mice than in the controls. The DNA content in the pallium cerebri and brain stem showed no increase and that of the cerebellum was even lower in quaking mice than in the controls. Hypertrophy of the astroglia was observed in the white matter of the cerebellum of quaking mice, though Bergmann astroglial fibers in the molecular layer did not show any hypertrophy. The cerebella of quaking mice in primary culture showed very poor myelination under the phase-contrast microscope. However, Purkinje cells from the quaking mutants appeared normal with regard to Bodian silver impregnation, hematoxylineosin staining, and Purkinje cell specific P 400 protein. Addition of the conditioned culture medium of qk/qk to the culture of the control cerebellum did not interfere with the myelination. We concluded that the cause of dysmyelination in the quaking mouse could be a genetic defect of the oligodendroglia rather than hypertrophy of the astroglial cells.


Biochemical characterization of postsynaptically localized cyclic nucleotide phosphodiesterase. This study demonstrates the postsynaptic localization of one of the isozymes of cyclic nucleotide phosphodiesterase (PDE) activity at asymmetrical, axospinous terminals in the rat corpus striatum and neocortex. Characterization of this enzymatic activity demonstrates that the PDE form surviving aldehyde fixation for electron cytochemistry can be considered to preferentially hydrolyze cyclic 3'5'-guanosine monophosphate, and it requires calcium and a heat-stable calcium-dependent regulator protein (CDR) for full hydrolytic activity. Ion exchange chromatographic analysis of extracts of corresponding unfixed brain regions demonstrates that only one enzyme activity peak exhibits similar aldehyde resistance and calcium and regulator protein activatibility.


Cation transport and membrane potential properties of primary astroglial cultures from neonatal rat brains. This paper describes K+ and Na+ content and transport in primary monolayer cultures from dissociated newborn rat brains, considered to consist predominantly of astroglial cells. Net changes in cation content after addition of ouabain, and steady state fluxes using 86Rb+ as a marker for K+ and 22Na+ as a marker for Na+, were measured. The results found indicate that the cells maintained a conventional pattern of cation homeostasis with net efflux of K+ being balanced by its active uptake and net uptake of Na+ balanced by active extrusion mediated by a ouabain sensitive (Na+K) pump. These processes maintained internal measured K+:Na+ ratios of 12--25:1. The cells were normally flat but addition of DBcAMP caused them to round up and form numerous processes, an appearance resembling that of astroglial cells in vivo. DBcAMP treatment also reduced the steady state levels of K+ measured with 86Rb+ by 15--30%, and had no effect on initial rates of 86Rb+ and 22Na+ uptake. The membrane potentials of cells treated with DBcAMP were studied, since only these were easily impaled. The membrane potentials of separate groups of cells gave means ranging from --65 to --75 mV at 35 degrees C, at an external K+ concentration ([K+]o) of 4.5 mM. The dependence of the membrane potentials of individual cells and groups of cells on [K+]o was studied. The slope of the potential per 10-fold change in [K+]o was 55--57 mV, at concentrations of K+ greater than 10--20 mM K+, and diverged from this slope at concentrations below this. This shows that these cells had some permeability to ions other than K+. Assuming that Na+ was the only other ion affecting the membrane potential, it was calculated that the permeability to Na+ was about 30 times less than K+. A similar result was obtained based on estimates of Na+ and K+ permeability from transport experiments on cells also treated with DBcAMP. The results obtained from these cells are compared to those found for other cultured glial cells and glial cells in vivo. We conclude that the membrane potentials of the cultured cells used in the present study show the closest resemblance so far to glia in vivo, since they are large and negative and are determined mainly by K+. However, the cultured cells have different properties from those reported in some studies for glial cells in vivo by showing free permeability to ions other than K+.


[Adaptation of blood sugar regulation during individually adjusted muscular exercises. Effects of training]. A twenty minutes exercise at 50 to 70% of VO2 max is followed by an increase of glycemia and F.F.A. Moderate physical activities could improve a better use of F.F.A., perhaps preferably to the glucose. Cortisol and A.C.T.H. levels raise, principally by the untrained subjects; however, that exercise is followed by a raise of STH, no dependent to the training's degree.


[Arterial blood pressure and high calcium diet in normal and mineralcorticoid (DOCA and sodium chloride hypertensive rats]. High calcium diet induces an hypertension lasting one week in normal rats. In mineralocorticoid treated rats (DOCA + NaCl), the same diet prevents for 10 weeks the increase of arterial blood pressure. Parathyroid activity (estimated by urinary cAMP) is decreased after the high calcium diet. These results confirm the role of the parathyroid glands in mineralocorticoid hypertension in the rat.


Pseudorabies: adaptation of the countercurrent immunoelectrophoresis for the detection of antibodies in porcine serum. A countercurrent immunoelectrophoresis test was developed for the detection of precipitating antibodies to pseudorabies virus in pig serum. The precipitation reaction occurred only between the pseudorables antigen and the homologous porcine antiserum. The sensitivity of the method was compared to that of the serum neutralization test. On the basis of its sensitivity, its specificity and the rapidity with which the results were obtained, the countercurrent immunoelectrophoresis may become a potentially valuable screening method to test large numbers of porcine serum.


Effect of temperature, relative humidity and medium on the aerosol stability of infectious bovine rhinotracheitis virus. Aerosols of infectious bovine rhinotracheitis virus were generated with a Devilbiss 40 nebulizer from Eagle's minimum essential medium, nasal secretion from a noninfected calf and nasal secretion from a calf artificially infected with infectious bovine rhinotracheitis virus and aged in a rotating drum at temperatures of 6 degrees C or 32 degrees C and relative humidities of 30% or 90%. The aerosols were sampled at seven minutes after start of spraying, one hour, two hours and three hours with an all glass impinger (AGI-30) and titrated for infectivity in cell cultures. Physical decay was determined by a rhodamine B tracer technique. During spraying (seven minutes from start of spraying), the virus was usually more stable in aerosols of nasal secretion from a noninfected calf and at 90% relative humidity. In nasal secretion from a noninfected calf the virus survived best at 90% relative humidity when the temperature was 6 degrees C and best at 30% relative humidity when the temperature was 32 degrees C. During aging, biological decay was greater at the higher temperature, and at 6 degrees C, the highest decay rates occurred at 30% relative humidity in Eagle's minimum essential medium and at 90% relative humidity in nasal secretion from a noninfected calf. The stability of infectious bovine rhinotracheitis virus infected nasal secretion was not widely different from that in noninfected nasal secretion, although under certain conditions greater survival occurred in the noninfected secretion.


Antibodies to bovine leukemia virus in a leukosis dairy herd and suggestions for control of the infection. A closed herd of 765 Holstein-Friesian dairy cattle with a history of multiple cases of leukosis was tested for antibodies to bovine leukemia virus by the bovine leukemia-glycoprotein immunodiffusion test. A total of 355 animals (46.4%) were antibody positive. Prevalence was 60% in the 373 milking cows and 100% in the breeding bulls. Antibodies were present in 59% of newborn calves. Prevalence of antibodies was higher in older animals and cows in second lactation had a higher prevalence than cows in first lactation (72% vs 43%). Proposed control measures in this herd aim at preventing infection of calves, heifers and lactating cows by: 1) separating them into groups negative and positive for bovine leukemia virus antibodies, 2) not allowing calves to receive colstrum or milk from infected cows and 3) by using seronegative bulls for natural breeding tested at three month intervals. Calves should be tested after six months of age. Before this time calves of positive mothers should be treated as being positive.


A rapid plaque neutralization test for bluetongue virus. A bluetongue virus plaque neutralization test, done in small tissue culture wells (2.0 cm2) using minimal antibody-virus contact time, was found to be of comparable sensitivity to more classical methods involving preincubation. By using the cell culture plates as serum dilution wells and adding cells directly to the virus-antibody mixture, a considerable saving in time is realized. Because of the small size of the wells, reagent cost and incubator space requirements are reduced making the test much more attractive for wide scale use, either as a screening test or one that yields dose-response lines.


Isolation of cloned Syrian hamster insulinoma cell lines with limited capacity for insulin production. Cell lines derived from a Syrian hamster insulinoma have been cloned in agar and maintained continuously in culture in vitro for 1 1/2 years. The cell lines have average doubling times of 48 h, they have modal chromosome numbers between 38 and 39, and they retain the ability to form tumors when injected into Syrian hamsters. The cells do not produce immunoreactive insulin when grown in vitro (less than 0.5 ng/mg protein), but do produce immunoreactive insulin when grown in vivo as tumors (mean value from six determinations = 7.1 +/- 1.5 ng/mg protein).


Investigation of a family suspected of being at high risk for cancer. The investigation of the family of a patient with bilateral breast cancer is described. By means of interviews and the checking of hospital records and death certificates, information was obtained on 199 family members over five generations, 19 of whom had cancer. Comparison with expected numbers of cases showed an excess in only one generation. The interpretation of these findings and the advice given to family members are discussed.


[Isolation of infectious bovine rhinotracheitis virus from the tarsal joint of a bull (author's transl)]. Isolation of infectious bovine rhinotracheitis virus from the tarsal joint of a bullInfectious bovine rhinotracheitis virus was isolated from the right tarsal joint of a bull who was showing signs of a systemic viral infection. The clinical signs manifested by 16 bulls of this herd are described, the laboratory methods used are listed and the results are analysed and discussed.


The significance of concordance in mammographic interpretations. Twenty-eight women participating in a screening program had a breast cancer diagnosed subsequent to the report of a negative mammogram which was read by only one of a group of radiologists. Fourteen occurred prior to a scheduled routine screening visit (interval cancers) and 14 were detected during such an examination. The negative mammograms from the 28 cancer patients, together with those from 120 women without cancer (controls) were independently reviewed by each member of a panel of three radiologists. Forty-six percent of the cancer cases and 5.8% of the controls were interpreted as positive by two or more of the radiologists. These findings suggest that agreement among several independent reviewers enchances the value and accuracy of mamography by reducing the number of false negative interpretations.


Nodular lymphoma with intracellular immunoglobulin. A case of malignant lymphoma with vacuolated cells is presented and, as with two other recent reports, is of the nodular poorly differentiated lymphocytic type. This form of malignant lymphoma presumably is composed of neoplastic B lymphocytes, which produce intracellular immunoglobulins or fractions thereof, corresponding to various cellular vacuoles and inclusions. The vacuolated appearance of the neoplastic cells simulates mucinous or "signet ring" adenocarcinoma, which may be excluded by clinical, routine microscopic, histochemical, immunologic, and electron microscopic observations.


Immunotherapy with levamisole: early decrease of cAMP levels in lymphocytes from treated cancer patients. Lymphocyte cyclic nucleotide content was studied before and after Levamisole administration to cancer patients. Twelve patients with disseminated cancer received 100 mg/m2 on two consecutive days; nine comparable patients with disseminated cancer served as controls. Endogenous cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) were measured in lymphocytes before, 24, and 48 hours after ingestion of the first dose of Levamisole. A statistically significant decline in lymphocyte cAMP level was observed after drug administration and no significant changes were noted in cGMP levels. Further studies will be necessary to correlate this biochemical change in cyclic nucleotides with modulation of the functional level of cellular immune mechanisms.


Multinucleated mammary stromal giant cells: a benign lesion that simulates invasive carcinoma. Multinucleated stromal giant cells were found in breast tissue from 14 patients treated by mastectomy for carcinoma over 3 decades (1950s to 1970s). Nine cases were found in a review of 200 consecutive mastectomies (4.5%) performed in 1978. The average age was 51 years with nine patients in their 40s. Other than the age distribution, no association with marital status, family history of breast carcinoma, hormone usage, tumor type, or stage of disease was observed. This benign lesion of undetermined significance should not be misinterpreted as invasive carcinoma. Multinucleated stromal giant cells have been observed in breast specimens from patients not known to have carcinoma at the time of biopsy. These patients had a similar age distribution as those with concurrent carcinoma. Follow-up will be required to determine if the association with carcinoma is significant or merely a coincidence reflecting age or some other yet to be identified factor.


Tumor-specific DNA sequences in human gliomas. Utilizing the technique of hydroxyapatite chromatography, normal cellular DNAs were used to recycle off the repeat or normal sequences found in [3H]DNA copied off 70S RNA from malignant astrocytomas. The recycled [3H]DNA were then used to hybridize against DNAs from normal human brain tissues and DNAs from malignant astrocytomas or Grade IV astrocytomas. The results indicated the presence of tumor-specific DNA sequences in malignant astrocytomas, absent in normal brain tissues. The percentages were 88% and 7%, respectively. When recycled medulloblastoma 70S[3H]DNA probes were utilized against DNA'S FROM Grade IV astrocytomas and from normal brain, similar results were obtained. The respective percentages of hybridization were 67% and 7%. Thus it would appear that malignant gliomas contain tumor-specific DNA sequences which are not found in normal brain tissues.


Primary malignant giant cell tumor of bone: a study of eight cases and review of the literature. Eight cases of primary malignant giant cell tumor of bone were reviewed. There was a wide range in age from 17 to 76 years, with the sixth decade of life being the most common. The tumor was more frequent among females (male to female ratio--3:5). The most common sites of occurrence were in the region of the knee, with the distal end of femur and the proximal end of tibia affected in three and two cases, respectively. Pain and swelling of the involved regions were the most common complaints. The roentgenographic and pathologic features and the treatment were analyzed in detail. Although these cases were considered malignant, the follow-up periods varying from 4 to 15 years were available in six of the eight cases; only one patient died of tumor, 8 months after the surgical procedure. One patient died of unrelated cause, but the others were all alive with no evidence of disease. The pertinent literature was analyzed and examples of secondary malignant giant cell tumors of bone were compared to those of this present series to delineate differences in natural history and clinicopathologic features. It was clearly established that primary malignant giant cell tumor of bone is a separate entity with a more favorable clinical behavior, particularly if the disease process is eradicated early on either by cryosurgery, en bloc radical resection, or amputation.


Elevation of brain-type creatine kinase in serum from patients with carcinoma. The brain isoenzyme of creatine kinase (CK BB) occurs in trace amounts in normal serum and is moderately increased in only a small number of non-oncological conditions. Although many tissues and tumors contain CK BB, we observed serum elevations only in certain carcinomas. Eleven patients with tissue-proven small cell anaplastic carcinoma (SCAC) of the lung had striking elevations of serum CK BB and no evidence of central nervous system (CNS) metastases. Significant increases were also observed in three cases of prostatic carcinoma with no apparent CNS involvement, and in one case each of adenocarcinoma and SCAC of the lung with proven CNS metastases. Three patients with SCAC of lung without distant metastases, three with SCAC of the esophagus with distant metastases but no known CNS involvement, and 17 patients with oncological conditions other than SCAC of the lung or adenocarcinoma of the prostate have failed to show elevation in serum CK BB activity. Serum CK BB may be useful as a diagnostic marker or indicator of metastases for some carcinomas.


Interaction of factors associated with cancer of the nasopharynx. A retrospective study of nasopharyngeal carcinoma (NPC) revealed that smoking, working under poor ventilation, use of nasal balms or oil for nasal and throat troubles, use of herbal drugs, and anti-EBV antibody titer were found statistically associated. The dural interactions of these factors to the risk of NPC were presented. Except in work conditions with poor ventilation and when herbal drugs are used, all the combinations were synergistic. The synergistic actions were especially remarkable with smoking and other factors. The possible etiological mechanisms of NPC are discussed.


The histology of liver tumors in oral contraceptive users observed during a national survey by the American College of Surgeons Commission on Cancer. As part of the national survey on the tumorigenesis of oral contraceptive drugs conducted by the American College of Surgeons' Commission on Cancer, a histologic study was made of 94 cases of liver tumors in users and non-users of oral contraceptives. Pathologic criteria were established and then the slides were studied; the results were tabulated to determine the significance of each of the criteria as related to the use of contraceptives. These criteria included tumor size, peliosis hepatis, hemorrhage, necrosis, fibrosis, thrombosis, and vascular alterations of the intima and media. Cases of focal nodular hyperplasia in pill users exhibited greater vascular alterations, fibrosis, peliosis, and tumor size as compared to focal nodular hyperplasia observed in non-pill users. In addition, hemorrhage, necrosis, and peliosis were much more common in hepatic cell adenoma than in focal nodular hyperplasia. In the material reported in this series there were no hepatic cell adenoma cases observed in non-pill users. Focal nodular hyperplasia cases exhibited an overwhelmingly greater degree of vascular intimal and medial alterations than hepatic cell adenoma. The results suggest that the effects of oral contraceptives on the liver may be primarily upon the vasculature.


Brain metastases in disseminated germinal neoplasms: incidence and clinical course. From July 1973, to March 1978, 139 patients with disseminated germinal neoplasms were treated with a variety of combination chemotherapy regimens. Twenty-one patients (15.1%) exhibited evidence of brain metastases at some time during their course. Central nervous system spread occurred somewhat more frequently in patients with choriocarcinoma and yolk sac histologies, although all types were at risk. This was a major cause of morbidity and mortality. However, central nervous system spread almost always occurred in patients with systemic relapse and thus did not affect overall survival. We have yet to identify a subgroup who potentially could benefit from prophylactic central nervous system therapy.


Familial islet cell tumors in Von Hippel-Lindau's disease. Von Hippel-Lindau's Disease is an hereditary disorder characterized by the development of hemangioblastomas of the cerebellum and retina and a variety of cystic and neoplastic lesions of other organs such as renal cell carcinoma and pheochromocytoma. In a single generation of a family with Von Hippel-Lindau's disease, all four siblings developed lesions classically associated with the complex. Additionally, two of the four developed islet cell tumors of the pancreas, one in one patient and five in the other. While a familial incidence of islet cell tumors is known in multiple endocrine adenomatosis, type I and Zollinger-Ellison syndrome, such a familial occurrence has been heretofore unrecorded in the Von Hippel-Lindau complex.


Unusual presentations of carcinoma of the right colon. Two unusual presentations of carcinoma of the right colon are described. One of the two patients presented with a right gluteal abscess and the other presented with a colocutaneous fistula to the anterior abdominal wall at the site of a previous appendectomy scar. Despite extensive local tumor invasion of adjacent structures in both cases, all the regional lymph nodes were free of cancer and the tumors were of low-grade malignancy. A review of the literature indicates that bulky colonic tumors with extensive local invasion and negative mesenteric lymph nodes have a relatively good prognosis if adequate resection is performed.


Adriamycin, BCNU plus cyclophosphamide (ABC) in advanced carcinoma of the head and neck. Forty-four patients with advanced carcinoma of the head and neck were treated with cyclophosphamide, 400 mg/m2 plus BCNU, 100 mg/m2 day 1 followed by adriamycin 40 mg/m2 day 2, with therapy repeated every 4 weeks. Of 31 evaluable patients, there were 1 complete response, and 10 partial responses (35%). Four of 8 patients without prior chemotherapy had complete or partial responses, as did 7 of 23 patients who had received prior chemotherapy. The mean survival for patients with partial responses was 8.8 months, and for patients with stable disease was 7.8 months. The mean time to progression for patients with partial responses was 5.4 months, and for patients with stable disease was 3.2 months. Granulocytopenia was the dose limiting toxicity, and patients with increasing degrees of myelosuppression appeared to have higher quality responses and longer survival. The ABC treatment program is useful in the palliative management of patients with advanced carcinoma of the head and neck.


Immunohistopathologic study on carcinoembryonic antigen (CEA)-like material and immunoglobulin A in gastric malignancies. Immunofluorescent staining for carcinoembryonic antigen (CEA)-like material and immunoglobulin A (IgA) was performed on 54 gastric specimens showing normal mucosa and various lesions including malignancy. Both CEA-like material and IgA were demonstrated in normal mucosa and benign and malignant epithelial lesions. Abnormal findings in respect to CEA-like material included staining of cell membranes with disordered pattern and increased cytoplasmic staining. Several staining characteristics for CEA-like material were shown by various types of malignant epithelial tumors. This staining provides a means of distinguishing anaplastic carcinoma from histiocytic lymphoma, as the latter does not demonstrate CEA-like material. IgA was consistently absent in anaplastic carcinomas, but was noted on the inner surface of tubular components in the differentiated adenocarcinomas, indicating functional preservation in the differentiated tumors. Immunohistochemical staining for CEA-like material and IgA is considered to be useful in the structural and functional study of gastric malignancies.


Intracytoplasmic type A virus-like particles in angioimmunoblastic lymphadenopathy. Virus-like particles which seemed to be oncogenic have not been observed clinically in human neoplasms. However, we found some virus-like particles in the cytoplasm of a few proliferating cells in the lymph nodes of one patient with angioimmunoblastic lymphadenopathy. These were very similar to intracytoplasmic type A particles seen in the Mazon-Pfizer monkey virus and a mammary tumor virus in a mouse.


Liver cell dysplasia: association with hepatocellular carcinoma, cirrhosis and hepatitis B antigen carrier status. Liver cell dysplasia was noted on histological examination of nontumorous liver from 24 of 50 (48%) black southern African males with hepatocellular carcinoma (HCC). Macronodular cirrhosis was present in 40 (80%). There was no statistically significant difference between the frequency of dysplasia in 50% of 40 cirrhotic and 40% of 10 noncirrhotic livers, or in 52.6% of 38 hepatitis B antigen (HBAg) positive and 33.3% of 12 HBAg negative HCC patients. HBAg positivity was present in 80% of 40 cirrhotic and in 60% of 10 noncirrhotic HCC patients. This lack of significant correlation between liver cell dysplasia, and both cirrhosis and HBAg positivity in HCC patients in contrast to findings in Uganda and the United States, suggests a different pathogenetic mechanism for dysplasia in southern Africa. Liver cell dysplasia in man appears to be analogous to preneoplastic experimentally-induced hyperplastic foci or areas.


Histologic prognostic indicators in hepatocellular carcinoma. Survival and histologic features were studied in 80 patients with proven hepatocellular carcinoma (HCC). The overall survival was 50% at 3.5 weeks with a statistically significant drop in mortality rate after 8.5 weeks. Patients with diffuse clear-cell pattern, focal clear-cell pattern, and no clear cells in their tumors had a 50% survival rate at 14 weeks, 6 weeks, and 3 weeks respectively; the differences in survival rates were statistically significant between any pair (p less than 0.001). No correlation was found between survival and (1) cytologic differentiation, (2) histologic architecture, (3) degree of pleomorphism, or (4) presence of bile production, proteinaceous secretion, giant cells, or cytoplasmic hyaline bodies. Eosinophilic HCC with lamellar fibrosis was not observed in any of our materials. We concluded that the presence of clear cells was the only histologic feature of prognostic significance in our patients.


Mucinous sweat gland adenocarcinoma of eyelid: a clinicopathologic study of 21 cases with histochemical and electron microscopic observations. This is a clinicopathologic study of 21 cases of mucinous sweat gland adenocarcinoma involving the eyelid. The tumor occurred in middle-aged adults (median age 60 years) and has a predilection for males. In this series eight patients (40%) had one or more local recurrences, one of whom died with extensive local invasion of the face after multiple recurrences in a 15-year interval. Only one patient had metastasis to the submandibular lymph nodes, which was treated by radical neck dissection. Involvement of the head, especially the face and scalp, was observed in 78% of the cases from the literature. Combining our cases with those previously reported, we found that the eyelid was involved in almost half of the cases (21 out of 45 lesions). We believe that the identification by light microscopy of a mixed population of light and dark secretory cells within the tumor lobules, the histochemical findings (presence of sialomucin and absence of iron as well as periodic acid-Schiff positive, diastase-resistant granules), and the results of ultrastructural studies support the histogenetic postulate that this mucinous adenocarcinoma is probably derived from the secretory cells of the eccrine coil. Our interpretation concurs with Headington's view of origin from the eccrine glands.


Plasma and tumor ACTH in carcinoma of the lung. Plasma and tissue contents of immunoreactive ACTH were determined in 100 patients undergoing surgical resection for lung cancer. ACTH was detectable (greater than 1 ng ACTH equivalent/g wet weight) tissue in 47 of 49 specimens of epidermoid carcinoma, in 15 of 17 specimens of adenocarcinoma and in 7 of 8 specimens of large cell carcinoma; the median concentrations in these three tumor types were 8, 3 and 9 ng/g, respectively. Patients with oat cell carcinoma were not considered candidates for curative resection and are not included in this series. ACTH was not detectable in 36 specimens of apparently normal lung tissue from the same lobe but distant from the tumor and ranged up to 6 ng/g in 14 other specimens. One-half of patients with epidermoid carcinoma but only one-quarter of those with adenocarcinoma had preoperative plasma levels exceeding 250 ng/L plasma. About 75% of patients survived for at least one year whether preoperative plasma levels were greater than 400 ng/L or less than 200 ng/L. In only 4 of 21 patients with preoperative levels greater than 300 ng/L did the plasma fall by at least one-half in the immediate postoperative period. It is concluded that measurement of preoperative and postoperative plasma immunoreactive ACTH does not have a prognostic value for remission or long term survival in lung cancer, probably because it is unlikely that the tumor per se is the sole source accounting for the elevated levels generally observed in this condition.


Malignant fibrous histiocytoma of the lung. Two malignant fibrous histiocytomas arising primarily in the lung are described. The first was a large tumor of the right lower lobe in a 53-year-old man. The other tumor was found incidentally on routine roentgenograms in a 25-year-old woman and involved the left main pulmonary artery. The lesions could be resected but both patients developed early cerebral metastases. The neoplasms were predominantly fibroblastic, had a characteristic storiform pattern, and included large histiocytes with bizarre nuclei and a vacuolated cytoplasm. The ultrastructure of the cells in the fibroblastic areas was characterized by irregular nuclei and a cytoplasm with a well-developed endoplasmic reticulum and dilated cisternae. Some cells lacked the prominent endoplasmic reticulum of fibroblasts and others were characteristic histiocytes with numerous cytoplasmic lysosomes. The cases appear to be the first reported primary malignant fibrous histiocytomas of the lung.


Argyrophil cell carcinoma of the uterine cervix with ectopic production of ACTH, beta-MSH, serotonin, histamine, and amylase. The case of a 38-year-old female with primary argyrophil cell carcinoma of the uterine cervix is reported. Two years after operation the patient developed widespread metastases with typical Cushing's syndrome. Microscopically, the tumor consisted of solid anaplastic cells, adenocarcinoma, and squamous cells. The plasma levels of ACTH and cortisol were elevated. Many cells of both the primary and metastatic tumors showed argyrophilia. Almost all the cells of the metastases contained numerous round secretory granules measuring about 117 micrometers in diameter. Small rod-shaped or larger round secretory granules, measuring 250 and 430 micrometers respectively, were also found in a few of these cells. The tumors in the right lung, pancreas, and kidney contained high levels of ACTH, beta-MSH, serotonin, histamine, and amylase. This is the first report of ectopic production of these five substances from argyrophil cell carcinoma of the uterine cervix.


Mucin-producing carcinoid ("composite tumor") of upper third of esophagus: a variant of carcinoid tumor. An unusual upper esophageal polypoid tumor was found to be predominantly a carcinoid tumor and in part a mucin-producing adenocarcinoma. A review of literature showed this to be a unique tumor in this location.


Remission of pure red cell aplasia associated with nonthymic malignancy. Two patients with acquired pure red cell aplasia associated with malignancy are presented. One patient has breast cancer and the other had poorly differentiated nodular lymphoma; neither patient had evidence of a serum inhibitor of hemoglobin synthesis and both achieved complete hematologic remission following prolonged immunosuppressive therapy. The literature describing the association of pure red cell aplasia and nonthymic malignancy is reviewed and potential for responsiveness discussed.


Inactivation of membrane-bound (Na+ +K+)-ATPase of Yoshida sarcoma cells and cobra venom cytotoxin complex with the glycolipid components of the enzyme system. The molecular mechanisms involved in the inactivation of (Na+ + K+)-stimulated ATPase of Yoshida sarcoma cells by a cytotoxic protein (P6) from cobra venom have been examined. The overall data obtained using purified (Na+ + K+)-stimulated ATPase of Yoshida sarcoma cells suggest that cytotoxin P6 combines with phosphatidyl serine and a glycolipid which are closely associated with (Na+ + K+)-stimulated ATPase which in turn may lead to the inactivation of the enzyme in this cell system.