The equilibrium constant and the reversibility of the reaction catalysed by nicotinamide-adenine dinucleotide kinase from pigeon liver. The reversibility of the NAD+ kinase reaction was established, and the kinetic parameters of the rate equation in the reverse direction were determined. The equilibrium constant of the reaction was determined by using the purified pigeon liver enzyme and radioactively labelled nicotinamide nucleotides. The relationship between kinetic parameters of the forward and reverse reactions is in reasonable agreement with the measured equilibrium constant. As expected from the proposed mechanism of action, the enzyme does not catalyse isotope exchange between NAD+ and NADP+ in the absence of ADP and ATP. Although homogeneous as judged by polyacrylamide-gel electrophoresis, the enzyme preparation exhibits ADP/ATP isotope-exchange activity which could not be separated from NAD+ kinase activity, but kinetic evidence suggests that this is probably due to a contaminant.


The effect of cyclophosphamide on MSV-H oncogenesis. The effect of cyclophosphamide on MSV-H oncogensis and the immune response of young mice has been investigated. A single, sublethal dose (100 and 50 mg/kg of cyclophosphamide) in 8-day-old mice given 24 h before or after MSV-H infection led to an earlier and lower incidence of tumours in comparison with controls infected only with MSV-H. The protective effect of cyclophosphamide, and the mechanism of action of both cyclophosphamide and MSV-H on the target cells, mesenchymal cells in rapid replication, as well the immunological implications of the findings are discussed.


Assessment of inherent fluctuations of mitotic and labelling indices of human tumours. A method is presented to evaluate the influence of statistical errors and inherent variation on the determination of mitotic and labelling indices of human tumours. In most of the experiments reported here, sufficient cells were counted to yield a statistical error which is small in comparison to the inherent differences in the proliferative indices, both between different sites in the same tumour and between different tumours of the same histological type. These inherent fluctuations are, theefore, a critical factor in cell kinetic studies of human tumours.


Chromosome complement and SV40 transformation of cells from patients susceptible to malignant disease. A comparative study has been made of fibroblasts obtained from patients with differing susceptibilities to malignant disease, both with respect to their chromosome complements and their transformation with SV40 virus. Fibroblasts from 2 Bloom's syndrome patients were found not to have raised SV40 transformation rates and no correlation was found between chromosome abnormality per se and transformation. Of 2 cell types with greatly increased rates, one was derived from a neurofibromatosis patient and the other from an A-T heterozygote. When SV40 DNA was employed as the transforming agent for the latter, the transformation rate was no longer raised.


Secretion of prostaglandins as bone-resorbing agents by renal cortical carcinoma in culture. Fragments of human renal carcinoma tissue have been co-cultured with mouse calvaria. In 9/13 cases significant bone resorption occurred whilst in no case did control kidney cause significant resorption. When bone resorption did occur, it could be reduced by inclusion of indomethacin in the culture medium. In some cases when theophylline was included in culture medium to prevent cyclic AMP breakdown, there was enhancement of tumour-induced bone resorption. Control studies without tumour showed that none of the experimental treatments had a direct effect on bone. Radioimmunoassay of prostaglandin E (PGE) levels in pooled culture media showed that tumour fragments produced appreciable amounts of PGE, and that this production was lowered by indomethacin and increased by theophylline. It is concluded that the bone resorption induced by these tumours is due to a prostaglandin, and that prostaglandin production may be controlled by changes in cyclic AMP metabolism.


The in vivo effects of quartz on rat thoracic lymph nodes. The histological changes in the regional thoracic lymph nodes of rats exposed to silica dust by inhalation are recorded. A dual response is noted involving both inflammatory and immune mechanisms, resulting in a fibroblastic connective tissue reaction and a plasma-cell-macrophage interaction. It is proposed that the progressive silicotic lesions obstruct the lymphatic channels in the lymph nodes, thus interfering with the lymph drainage from the lung and aggravating the silicotic process in the lung itself.


EMC viral infection of the coronary blood vessels in newborn mice: viral vasculitis. The coronary arteries, veins and capillaries of newborn mice experimentally infected with encephalomyocarditis (EMC) virus were studied histologically and electronmicroscopically. Damage to these coronary vessels was patchy. Some lesions were extensive, particularly when adjacent to areas of myocardial damage. The histological findings consisted of swelling of adventitial and endothelial cells pyknosis, and polymorphonuclear leucocyte and round cell infiltration. Histological and ultrastructural changes occurred in all three layers of the vessels. EMC viral crystals were found electronmicroscopically in the adventitia of the coronary arteries and veins. Ultrastructural lesions typical of virocytonecrosis were frequently noted in association with viral crystals, establishing the viral etiology of the lesions. These findings lend support to the concept that the patchy atherosclerotic and arteriosclerotic lesions found in the coronary vessels of man may represent healed lesions of previous viral damage.


Specific immunity in patients suffering from recurring warts before and after repetitive intradermal tests with human papilloma virus. The specific humoral and cellular immunity of 22 patients with multiple or recurring warts was studied. After repeated intradermal tests, using an inactivated, purified viral antigen, the responses obtained could be classed into two groups. The first group (10 patients) was characterized immunologically by the acquisition of a specific cellular immunity and the appearance of circulating IgG antibodies, and clinically by a total regression of resolution of the warts two to three weeks after the final intradermal test. The second group (12 patients) was characterized immunologically by a weak or non-existent specific immune response, and clinically by the unmodified persistence of the warts.


A biochemical explanation for the fatty liver and kidney syndrome of broilers: its alleviation by the short-term use of dietary fat. 1. Fatty livers and kidney syndrome (FLKS) was induced in young broiler chickens by giving them a diet composed principally of wheat and meat meal. 2. FLKS resulted in reduced growth and increased liver weight; fasting for 18 h increased mortality, liver lipid and the specific activity of hepatic ATP-citrate lyase compared with birds fed on a commercial diet. The specific activities of hepatic fructose-1,6-diphosphate-1-phosphohydrolase and pyruvate carboxylase were reduced in birds suffering from FLKS and fasted for 18 h. 3. Feeding of the FLKS-inducing diet supplemented with 150 g animal tallow/kg for 54 h considerably reduced mortality while restoring liver composition and enzyme activities towards those observed in birds fed a commercial diet. Investigations indicated that the glycerol component of the fat was not responsible for the observed responses. 4. The present results suggest that in FLKS insufficiencies of biotin are induced in specific enzyme systems, but the syndrome may be alleviated without the use of supplementary biotin. 5. The evidence indicates that, when stressed, birds affected by FLKS diet from the hypoglycaemia occurring as a result of a reduced capacity for gluconeogenesis.


Maternal nutrition in early pregnancy.

Maternal nutrition in early pregnancy. 1. Mean daily nutrient intakes of 195 women in the first trimester of pregnancy were assessed by weighed dietary records. 2. In comparsion with recommended intakes for non-pregnant women aged 18-55 years (Department of Health and Social Security, 1969), more than two-thirds of the subjects were having insufficient energy, iron and cholecalciferol. Unsatisfactory intakes of other nutrients were not uncommon. In relation to recommended intakes for the second trimester (Department of Health and Social Security, 1969), all mothers were having insufficient cholecalciferol and more than 80% of mothers had unsatisfactory intakes of energy and Fe. 3. Intakes appreciably lower than those recommended were associated with the following factors: social classes III, IV and V; maternal age under 20 years; smoking ten or more cigarettes daily; vomiting on more than 3 d/week.


Nuclear magnetic resonance studies of lipid-protein interactions. A model of the dynamics and energetics of phosphatidylcholine bilayers that contain cytochrome c oxidase. Reconstituted membrane systems of synthetic phosphatidylcholines and the integral membrane enzyme cytochrome c oxidase were prepared in order to conduct nuclear magnetic resonance studies of lipid-protein interactions. These lipids, labeled with a geminate difluoro group on the 1-position hydrocarbon chain, were combined with the enzyme to give active lipid-protein particles with a well-defined ratio of lipid to protein. The fluorine magnetic resonance spectra of a series of preparations with different lipid/protein ratios suggest that the hydrocarbon chain mobility of the lipid is substantially reduced with increasing amounts of protein. The fluorine spectra of a single lipid-protein preparation show a dramatic increase in the number of the more mobile lipid chains with increasing temperature. The results suggest that the enzyme orders the lipid bilayer well beyond those lipids in direct contact with the protein surface, and that the amount of the lipid restricted by the enzyme is dependent upon temperature. The exchange of lipid between the restricted and the more mobile lipid environments most probably does not occur over the time scale measurable by the magnetic resonance techniques, about 10(-3) s.


Lipid requirements for cytochrome c oxidase activity. Cytochrome c oxidase depleted of endogenous lipid by detergent exchange has been reconstituted into vesicles with synthetic lipids of known head group and fatty acid composition and enzymic activities have been measured. No evidence for head group specificity was found. However, the enzyme does require the fluid environment provided by unsaturated fatty acids. The state of dispersion of the enzyme was found to affect the activities regenerated in reconstitution studies. The highest activities were obtained using lysolecithin containing an oleoyl fatty acid as the lipid component.


In vivo inhibition of Novikoff cytoplasmic messenger RNA methylation by S-tubercidinylhomocysteine. The analogue S-tubercidinylhomocysteine (STH) has been used to study the methylation of mRNA in vivo. Partial inhibition of cytoplasmic poly(A)-RNA methylation was observed using a level of inhibitor which still permitted cell growth. Characterization of the partially methylated mRNA indicated the presence of cap structures lacking 2'-O-methylnucleosides, m7GpppN', which are normally not found in mammalian mRNA. Inhibition of additional methylated sites in mRNA at the second 2'-O-methynucleoside, and at internal N6-methyladenosine was also observed Methylation of 7-methylguanosine was not affected under the conditions used in these experiments. The methylnucleoside composition of cap structures differed in STH-inhibited and uninhibited cells. These results indicate that a completely methylated cap is not required for transport of mRNA into the cytoplasm. Furthermore, it may now be possible to assess in vivo the sequential nature of mRNA methylation and its potential role in mRNA processing.


Changes in the plasma lipoprotein-apoproteins of guinea pigs in response to dietary cholesterol. The major apoproteins from four plasma lipoproteins were isolated from control and cholesterol-fed guinea pigs. Apoproteins were studied by column chromatography, polyacrylamide gel electrophoresis, and amino acid analysis. Dietary cholesterol altered the plasma apolipoproteins mainly by an enrichment in the content of arginine-rich polypeptide (ARP) in all density fractions. This protein had a similar molecular weight (34 000), electrophoretic mobility, amino acid composition, and microheterogeneity as ARP reported in other mammalian species. The estimation of plasma concentration of ARP indicates a higher correlation coefficient with plasma unesterified cholesterol (r = 0.98) compared with cholesterol esters (r = 0.62).


Neurotoxins of Bungarus multicinctus vernom. Purification and partial characterization. The purification to homogeneity of nine neurotoxic components of the venom of Bungarus multicinctus is described. The purified components include alpha-bungarotoxin and two other alpha-type synaptic toxins and beta-bungarotoxin and five other beta-type synaptic toxins. The purified toxins have been characterized by electrophoresis, isoelectric focusing, amino acid analysis, and N-terminal amino acid determination. The alpha-type synaptic neurotoxins constitute a discrete class with molecular weights of 7000-8500, isoelectric points (pI) of 9.0-9.2, and N-terminal isoleucine or methionine. The beta-type synaptic neurotoxins constitute a second group with molecular weights of 20 000-22 000 and pI = 8.8-9.7. Fractions 10 through 13 exhibit a chain structure consisting of a 6000-7000 light chain and a 11 000-15 000 heavy chain apparently covalently stabilized by interchain disulfides. Fractions 9A and 14 were single chains of 11 000-14 000 which resemble the sequenced beta-type synaptic neurotoxin notexin (Halpert, J., and Eaker, D. (1975), J. Biol. Chem. 250, 6990). All of the beta-type synaptic toxins have a single tryptophan and N-terminal aspartic acid or asparagine.


Purification of cyclic 3',5'-nucleotide phosphodiesterase inhibitory protein by affinity chromatography on activator protein coupled to Sepharose. The Ca2+-dependent, reversible, interaction of cyclic adenosine 3',5'-monophosphate (cAMP) phosphodiesterase with its activator has been used to purify the enzyme by affinity chromatography. Activator-dependent cAMP phosphodiesterase is only a minor component of the proteins specifically adsorbed in the presence of Ca2+ by the Ca2+-dependent activator protein coupled to Sepharose and subsequently released by [ethylenebis(oxyethylenenitrilo)]tetraacetic acid. The major protein component can be partially resolved from the enzyme by gel filtration on Sephadex G-200. This protein has been purified to apparent homogeneity and shown to be composed of two polypeptide chains with molecular weights of 61,000 and 15,000 respectively. This protein is, by itself, devoid of phosphodiesterase activity and inhibits the activation of cAMP phosphodiesterase by its activator without affecting the basal activity. Thus, activation of cAMP phosphodiesteriase by the Ca2+-dependent activator protein may be controlled by interactions with yet a third component of the enzyme complex.


Altering the specificity of restriction endonuclease: effect of replacing Mg2+ with Mn2+. In the presence of 100 mM Tris buffer (pH 7.5) and 1-10 mM Mg2+ EcoRI endonuclease cleaves DNA at a specific nucleotide sequence and in a characteristic way: -GAATTC-. But if Mg2+ is replaced by Mn2+, the specificity of the cleavage is relaxed and cleavages occur at many other sites; moreover, there appears to be a hierarchy of cleavage rates at the pseudo-EcoRI restriction sites. For example, SV40 DNA is cleaved only once in the usual digestion conditions, but with Mn2+ more than ten cleavages are made; the five most rapidly cleaved SV40 DNA map locations are 0/1.0 larger than 0.93 larger than 0.33 approximately equal to 0.42 larger than 0.29 approximately equal to 0.40 larger than 0.25. Mn2+ also alters the restriction specificity of HindIII but not HpaII endonuclease.


Sedimentation behavior of native and reduced apolipoprotein A-II from human high density lipoproteins. The solution properties of human serum apolipoprotein A-II, both in the native and in the reduced forms, were investigated by the technique of sedimentation equilibrium in the analytical ultracentrifuge. For both proteins, the apparent weight average molecular weights determined in neutral buffer systems were found to be dependent on protein concentration and invariant with the rotor speeds used (16,000 to 44,000 rpm) indicating a reversible self-association. These results were also found to be independent of temperature between 5 and 30 degrees C. The pattern of self-association of native apolipoprotein A-II could best be described by a monomer-dimer-trimer equilibrium, in agreement with previously reported data (Vitello, L B., and Scanu, A. M. (1975), Biochemistry 15, 1161). The self-association pattern of apolipoprotein A-II reduced in the presence of 50 mM dithiothreitol conformed with a monomer-dimer-tetramer equilibrium similar to that reported for the native single chain apolipoprotein A-II of the rhesus monkey (Barbeau, D. L., et al. (1977), J. Biol. Chem. 252, 6745), but differing significantly from that reported for the reduced and carboxymethylated human product (Osborne, J. C. , et al. (1975), Biochemistry 14, 3741).


Estimation of transmembrane potentials from phase equilibria of hydrophobic paramagnetic ions. Positively charged hydrophobic spin labels have been synthesized which respond to transmembrane potentials in sonicated liposomes. Electron paramagnetic resonance spectroscopy is used to show that the distribution of these probes between aqueous and membrane phases changes as a function of transmembrane potential. When liposomes are made more inside-negative, the fraction of membrane associated probe increases while the fraction of probe in the aqueous phase decreases. The results are in quantitative agreement with a simple equilibrium thermodynamic theory which allows estimation of absolute transmembrane potentials in phospholipid vesicles.


Analysis of rat repetitive DNA sequences. Parameters of repetitive sequence organization have been measured in the rat genome. Experiments using melting, hydroxylapatite binding, and single strand specific nuclease digestion have been used to measure the number, length, and arrangement of repeated DNA sequences. Renaturation and melting or S1 nuclease digestion of 1.0 kbp DNA fragment show about 20% of rat DNA sequences are 3000-fold repeated. Renatured duplexes from 4.0 kbp DNA fragments display two repetitive size fractions after nuclease digestion. About 60% of the repeated sequences are 0.2-0.4 kbp long while the remainder are longer than 1.5 kbp. The arrangement of the repeated sequences has been measured by hydroxylapatite fractionation of DNA fragments of varying lengths bearing a repeated sequence. Repeated DNA sequences are interspersed among 2.5 kbp long nonrepeated sequences throughout more than 70% of the rat genome. There are approximately 350 different 3000-fold short repeated sequences in the rat interspersed among 600,000 nonrepeated DNA sequences.


Sequence organization of the rat genome by electron microscopy. The size and arrangement of repetitive and inverted repeat (foldback) sequences in rat DNA were studied by visualization of hybrid and heteroduplex structures in the electron microscope. The self-reassociation of repetitive sequence-bearing DNA strands often results in the formation of four-ended "H" structures, whose duplex regions equal the repetitive sequence length and can be measured in the electron microscope. In this way, it was determined that the average size of the class of numerous short repetitive sequences is 0.40 +/- 0.15 kbp. Heteroduplex structures were prepared between long whole DNA single strands and short repeat-sequence-bearing strands. The analysis of these structures confirms that the size of the repetitive sequences in 0.4 kbp on average. Length measurements between adjacent duplexes show that the average spacing between two interspersed repeats is at least 1.5-1.8 kbp. By examining 29.4-kbp single strands after brief renaturation, the size and distribution of foldback sequences were determined. There are 1.9 X 10(5) foldback apirs per rat genome, spaced an average of 9.7 kbp apart according to our measurement. Repetitive, inverted repeat and unique sequences are interspersed with each other in at least half the genome.


The mechanism of energy conservation and transduction by mitochondrial cytochrome c oxidase. Oxidation of ferrocytochrome c by molecular oxygen catalysed by cytochrome c oxidase (cytochrome aa3) is coupled to translocation of H+ ions across the mitochondrial membrane. The proton pump is an intrinsic property of the cytochrome c oxidase complex as revealed by studies with phospholipid vesicles inlayed with the purified enzyme. As the conformation of cytochrome aa3 is specifically sensitive to the electrochemical proton gradient across the mitochondrial membrane, it is likely that redox energy is primarily conserved as a conformational "strain" in the cytochrome aa3 complex, followed by relaxation linked to proton translocation. Similar principles of energy conservation and transduction may apply on other respiratory chain complexes and on mitochondrial ATP synthase.


Effects of deoxycholate and phospholipase A2 on choline and ethanolamine phosphotransferases of chicken brain microsomes. Ethanolamine phosphotransferase (EC 2.7.8.1) and choline phosphotransferase (EC 2.7.8.2) activities were assayed in fresh microsomes from adult chicken brains with either diacylglycerols or alkylacylglycerols. Pretreatment of microsomes with 1.25 mM sodium deoxycholate, a concentration less than the critical micelle concentration, produced a slight inhibition of choline phosphotransferase activity. A deoxycholate concentration (5.0 mM) greater than the critical micelle concentration (3.0 mM) decreased the choline phosphotransferase activity by more than 70% but had no effect on ethanolamine phosphotransferase activity. Inclusion of 1.25 mM deoxycholate in the assay medium decreased choline phosphotransferase activity 35% but increased ethanolamine phosphotransferase activity 50%. The deoxycholate appeared to inactive the choline phosphotransferase. Phospholipase A2 (Vipera russelli) treatments of microsomes removed phosphoglycerides and decreased both phosphotransferase activities to a similar extent. Decreased activities are probably due to disruption of the membrane structure. Choline and ethanolamine phosphotransferase activities are apparently in different enzymes which lack specificity for the type of diglyceride. Thus, the systematic names should include 1,2-diradyl-sn-glycerol instead of 1,2-diacyl-sn-glycerol.


NAD-dependent 3alpha- and 12alpha-hydroxysteroid dehydrogenase activities from Eubacterium lentum ATCC no. 25559. Eubacterium lentum (ATCC No. 25559) was shown to contain 3alpha-and 12alpha-hydroxysteroid dehydrogenases both of which were NAD-dependent and active against conjugated and unconjugated bile salts. In addition, the 3alpha-hydroxysteroid dehydrogenase was active against members of the Androstan series containing a 3alpha-hydroxyl group regardless of the stereo-orientation of the 5-H-. No measurable activity against 7alpha-, 7beta-, 11beta-, or 17beta-hydroxyl groups was demonstrated. The growth of E. lentum and the production of 3alpha- and 12alpha-hydroxysteroid dehydrogenases were greatly enhanced by the addition of L-, D- or DL-arginine to the medium. Yields of hydroxysteroid dehydrogenase were optimal in the range of 0.50-0.75% arginine; however, the growth of the organisms was further enhanced at arginine concentrations greater than 0.75%. The 12alpha-hydroxysteroid dehydrogenase was heat labile and could be selectively inactivated by heating at 50 degrees C for 45 min. Both the heated enzyme preparation (containing only 3alpha-hydroxysteroid dehydrogenase) and the unheated enzyme preparation (containing 3alpha- and 12alpha-hydroxysteroid dehydrogenases) were useful in the spectrophotometric quantification of bile salts. The optimal pH values for 3alpha- and 12alpha-hydroxysteroid dehydrogenases were 11.3 and 10.2, respectively. Kinetic studies have Km estimates of 2.10(-5) M and 1.0.10(-4) M with 3alpha,7alpha-dihydroxy-5beta-cholanoyl glycine and 7alpha,12alpha-dihydroxy-5beta-cholanoate for the two respective enzymes.


The turnover of human plasma very low density lipoprotein protein. The plasma decay of three groups of iodinated apoproteins on human very low density lipoproteins were evauluated in two normals, two subjects with endogenous hypertriglyceridemia and another two with dysbetalipoproteinemia. The apo beta decay was more rapid than that of the C apoproteins in all patients. The apo beta decay was more rapid for the normals than for either the subjects with hypertriglyceridemia or dysbetalipoprotenemia. The apo C protein had an irregular decay in the normals but decayed less irregularly for the hypertriglyceridemics. The arginine rich apoprotein had a decay somewhat similar to apo C protein in the normals. The apo beta protein of the alpha2 very low density lipoprotein of a dysbetalipoproteinemic was consistent with a precursor relationship to the apo beta of beta very low density lipoprotein of this subject, but the arginine rich apoprotein was not.


Subunit structure of the reconstitutively active cytochrome b-c1 complex. Determination of amino acids and molar distribution of subunit fractions from gel electrophoresis. A quantitative method has been developed to analyze the amino acid composition of protein subunits directly from the Coomassie Blue-stained band of polyacrylamide gel columns after electrophoresis. It is an improved method originally reported by Houston (Houston, L. L. (1971) Anal. Biochem. 44, 81--88). The results obtained can be thus used for the calculation of the molar ratios of subunit components of protein. The manipulation of the method and computation of the results are illustrated by a very complicated lipoprotein complex. The subunit molar ratios of the reconstitutively active cytochrome b-c1 complex were determined to be 2, 2, 2, 3, 2, 2, and 5 among the seven bands of the corresponding molecular weights of 53 000, 50 000, 37 000, 30 000, 28 000, 17 000, and 15 000, from gel electrophoretic columns. The amino acid composition of each subunit fraction determined directly from hydrolysis of gel was comparable with that obtained by actual isolation of each subunit.


Identification of subunits of bovine heart cytochrome oxidase. Purified lipid-depleted cytochrome oxidase, at purity of 12--14 nmol heme a per mg protein, has been shown to contain seven non-identical subunits in the ratio of unity. Their molucular weights on polyacrylamide gel are, in thousands, 40, 21, 14.8, 13.5, 11.6, 9.5, and 7.6 from gel electrophoresis after dissociation in sodium dodecyl sulfate and beta-mercaptoethanol. The molar ratio is determined by the amino acid composition of each subunit obtained from direct hydrolysis of the stained polyacrylamide gel slices. The amino acid composition of the isolated subunits I and II determined by regular hydrolysis method is found practically the same as that from direct hydrolysis of gel slices. The heme-associated polypeptides are identified with subunits of molecular weights of 40.10(3) and 11.6.10(3). One of the two coppers associated with the polypeptide of molecular weight of 21 000. The second copper may be associated with heme in the subunit of 40.10(3). Evidence of the existence of interpolypeptide disulfide linkages is presented.


Determination of calcium transport and phosphoprotein phosphatase activity in microsomes from respiratory and vascular smooth muscle. 1. Calcium transport into microsomal vesicles of respiratory (tracheal) smooth muscle was characterized. This calcium transport was ATP dependent and stimulated by the presence of the oxalate ion. The magnitude of transport was similar to that reported for microsomes from other types of smooth muscle. 2. Bovine and rabbit, heavy and light microsomes were isolated from respiratory (tracheal) and vascular (aortic) smooth muscle. Preincubation of these vesicles with cyclic AMP and protein kinase did not alter the transport of calcium into the vesicles. There uas no evidence of phosphate incorporation into microsomal membrane proteins. Similar results were obtained if phosphorylase b kinase replaced the combination of cyclic AMP and protein kinase during the preincubation. 3. The phosphoprotein phosphatase activity of cardiac sarcoplasmic reticulum and smooth muscle microsomes was determined. The activity of this enzyme was found to be several-fold less in the cardiac sarcoplasmic reticulum than in various smooth muscle microsome preparations.


Hemosiderin. an EPR study of water-insoluble iron in human and rat liver. EPR spectra of the water-insoluble iron fraction, hemosiderin of human and rat liver are described. The homogenate of freshly prepared perfused rat liver shows a non-heme iron signal at g=4.3 and a high-spin heme-iron signal around g=6, whereas the washed and sonicated sample of the insoluble iron fraction shows solely a non-heme iron signal at g=4.3. This indicates that hemosiderin from rat liver does not contain heme iron. Human-liver preparations from post mortem obtained material show in the homogenates as well as in the washed and sonicated samples an intense high-spin heme iron signal at g=6.0 and a non-heme iron signal at g=4.3. A comparative experiment, carried out with "aged" rat liver preparations, reveals the same spectra as with the human preparations. It is concluded that that the heme present in the insoluble iron fraction is caused by degradation of hemoglobin in the obduction material, and that heme is not a constituent of the insoluble depot iron.


Modified cyclic nucleotide systems in Morris hepatoma 3924A favoring expression of cyclic GMP effect. Modifications in the cyclic nucleotide systems favoring the expression of cyclic GMP effects were found to occur in the transplanted fast-growing Morris hepatoma 3924A. These included: (a) a decreased level of cyclic GMP phosphodiesterase and an increased level of cyclic AMP phosphodiesterase; (b) a disproportionately increased level of cylic GMP-dependent protein kinase relative to that of cyclic AMP-dependent protein kinase; (c) a disproportionately increased level of stimulatory modulator of cyclic AMP-dependent protein kinase relative to that of inhibitory modulator of cyclic AMP-dependent protein kinase; and (d) an increased level of phosphoprotein phosphatase.


Lysis of halophilic Vibrio alginolyticus and Vibrio costicolus induced by chaotropic anions. High concentration (1.0 M) of KSCN, but not of NaSCN, induced lysis of slightly halophilic Vibrio alginolyticus and moderately halophilic Vibrio costicolus, and the decrease in absorbance of the cell suspension was complete after 30 min at 25 degrees C. Replacement of K+ with Na+ effectively prevented the lysis by SCN-.K+ salts of NO3-, Br- and I-, however, induced no significant lysis. In electron micrographs, a prolonged exposure of the cells of V. alginolyticus to 1.0 M KSCN displaced the nucleoplasm to maintain close contact with the cell membranes. After 40 min of interaction, 50% of the cellular protein, 96% of RNA and 94% of DNA were recovered in the lysed cells. In contrast to lysis in hypotonic conditions, the lysis induced by KSCN is due mainly to a partial release of protein from the cells. V. costicolus  was more susceptible to SCN- than V. alginolyticus, whereas nonhalophilic Escherichia coli was resistant to 1.0 M KSCN. Thus, lysis by SCN- is characteristic of halophilic bacteria and cell membranes of more halophilic bacteria are more susceptible to chaotropic anions. The protective effect of Na+ observed here was considered to be manifested by specific interactions of Na+ with components of cell membranes, thereby rendering their structures resistant to the action of chaotropic anions.


[Spectral-kinetic characteristics of the cation-radical of pheophytin a]. The photochemical generation of pheophytin a cation-radical in acidulated ethanol and aceto nitril has been studied by the method of flash-photolysis. The bands with maxima at 450, 580 and 800 nm correspond to pheophytin a cation-radical in the differential absorption spectra. The efficient rate constants of pheophytin a triplet state desactivation in ethanol, acetonitril, aceton have the following values: 1.5.10(4), 9.10(3), 8.7.10(3) sec(-1) correspondingly. The rate constant of electron transfer from pheophytin a to p-benzoqui-none in ethanol solutions is 7.8.10(9) l/m.sec. The recombination constants of pheophytin a cation-radical with Q-in acetonitril, and with quinone neutral (QH) in acidulated ethanol have the following values: 5.10(9) and 2.10(9) l/m.sec correspondingly.


[Effect of coenzyme on conformational stability of glyceraldehyde-3-phosphate dehydrogenase from muscles of ecto- and endothermic animals]. The stabilizing effect of the coenzyme (NAD) on the structure of glyceraldehyde-3-phosphate dehydrogenase from lamprey and porcine muscles with respect to proteolysis and heat denaturation was studied. The process of heat denaturation was followed by the changes in specific activity of the enzymes; that of proteolysis--by the changes in specific activity and circular dichroism. It was shown that in both cases NAD at saturating concentration exerts a far weaker stabilizing effect on the structure of glyceraldehyde-3-phosphate dehydrogenase from lamprey muscle than on that of the porcine muscle enzyme. The coensyme-dependent stabilization of lamprey muscle glyceraldehyde-3-phosphate dehydrogenase does not differ from that of mammalian muscle enzyme. Possible interrelationship between the phenomenon observed and the molecular mechanism of thermal adaptation in the cold-blooded animals is discussed.


[Model of electrical feedback mechanisms through chemical synapses]. The model of excitatory chemical synapse is proposed in which the feed-back between the post- and presynaptic neurones is carried out by means of synaptic current generated by postsynaptic neurone. In the model, the potential drop generated by this current along the intercellular gap near the activated synapse (RIII) evokes additional depolarization of presynaptic membrane and thus stimulates the additional release of transmitter (fig. 1, A and B). The feed-back, which increases the steepness of transfer function between pre- and postsynaptic neurones is the more effective the higher the resistance of intercellular gap (fig. 2 and 3, B). is very low, so that activation of synapse is unable to change significantly its membrane potential (V2), the feed-back is the most effective, as revealed in additional depolarization of presynaptic membrane (VII in fig. 3, A). In inhibitory synapse the feed-back by electrical current is negative one.


[Properties of mitochondria from cells of the "fermentative" variant of Endomyces magnusii]. The properties of mitochondria from the cells of the "fermentative" variant of End. magnusii were studied. The induced fermentative transformation was brought about by a non-balanced vitamin cultivation. It was shown that the "fermentative" variant of End. magnusii represents an interesting model, in which the energy required for the cell functioning is provided for by a high fermentative activity and a normally functioning respiratory chain. The "fermentative" variant mitochondria were tightly coupled and possessed theoretical efficiency during oxidation of NAD-dependent substrates, which suggested the existence of all the three sites of energy coupling and phosphorylation at the substrate level. A specificity of energy regulation of the End. magnusii "fermentative" variant mitochondria, e. g. tight coupling during oxidation of succinate and lack of tight coupling during oxidation of exogenous NADH, is discussed. The tight coupling during succinate oxidation is confirmed by the observation of reverse electron transfer. Thus, the energy-dependent reduction of NAD during succinate oxidation has been firstly demonstrated for the mitochondria of yeast grown on a fermentable substrate.


[Lipid dependence of mitochondrial monoamine oxidase from rat hepatoma 27 with the use of rat liver lipid exchange proteins]. The ability of liver lipid exchange proteins to introduce foreign phospholipids into intact mitochondria was used for a study of the lipid dependence of monoamine oxidase. Introduction of exogenous phosphatidylcholine into rat hepatoma mitochondria, in which both the monoamine oxidase activity and the phosphatidylcholine content are comparatively low, leads to considerable activation of the enzyme. The introduction of exogeneous phosphatidylserine, phosphatidylethanolamine and cardiolipin has no activating effect. This indicates that the decreased activity of monoamine oxidase in the hepatoma may be due to a low amount of phosphatidylcholine. The method described allows the study in situ of the lipid dependence of non-solubilized membrane-bound enzymes.


[Chemical characteristics of conformationally-disequilibrium states of metal-containing proteins]. Properties of conformation-out-equilibrium states of several iron containing proteins, for example, induced by fast changes of active site were investigated. It was shown that structure and chemical properties of out-of-equilbrium states can differ considerably from the equilibrium ones. Spectral characteristics and reactional ability of these molecules change in the way of slow (up to 10(-1)s) conformational relaxation, continuously approaching the equilibrium values.


[Amino acid sequence of corticotropins from seiwhale (Balaenoptera borealis) and pinwhale (Balaenoptera physalus)]. The corticotropins from two species of whales, e. g. seiwhale (Balaenoptera borealis) and finwhale (Balaenoptera physalus) were subjected to hydrolysis by trypsin, chymotrypsin and pepsin. The peptide fragments were separated by gel-filtration through Sephadex and partition paper chromatography. The study of the amino acid sequence of the peptides obtained allowed to establish the primary structure of corticotropin from both species, which was found structurally identical to human corticotropin.


[Interaction of 8-substituted derivatives and adenosine-3',5'-cyclophosphate esters with protein kinase from pig brain]. A synthesis of previously unknown 8-substituted derivatives and alkyl esters of cyclic adenosine-3',5'-monophosphate, containing reactive groups, was carried out. The interaction of the compounds obtained with a homogeneous preparation of protein kinase from pig brain was studied. It was found that all compounds, with the exception of neutral esters of 3',5'-AMP, activate the enzyme and competitively inhibit 3H-labelled 3',5'-cAMP binding by the regulatory subunit of protein kinase. The activating effect and affinity of 8-(beta-aminoethylamino)-3',5'-cAMP for protein kinase was 10 times lower than that for 3',5'-cAMP and other 8-substituted derivatives of the cyclic nucleotide. It was found that 8-(N-chloroacetylaminoethylamino)-3',5'-cAMP interaction with the enzyme is of irreversible type, which suggest covalent blocking of the nucleophilic group of the 3',5'-cAMP binding site of protein kinase. The data obtained indicate that the 3',5'-cAMP molecule is bound to the regulatory site of protein kinase in the syn-conformation. The previously made assumption on the crucial importance of the negative charge in the 3',5'-cyclophosphate system for the interaction of cyclic AMP with the regulatory subunit of protein kinase has been thus confirmed.


Identification of microcrystals in synovial fluids by combined scanning electron microscopy and x-ray diffraction : application to triclinic calcium pyrophosphate dihydrate. We report a method for studying microcrystals which combines scanning electron microscopy, electron-dispersive X-ray analysis, and X-ray diffraction. Application of all three techniques to the same crystalline bodies permits correlation of their three-dimensional morphology at high magnification with unambiguous identification by means of their crystallographic properties. The method was applied to triclinic calcium pyrophosphate dihydrate.


Functional abnormalities of neutrophils in cancer patients : inefficient phagocytosis and reverse endocytosis. A study of neutrophil functions has been performed in 17 patients with advanced cancer and in 21 controls. Their phagocytic activity, their index of reduction of NBT and their myeloperoxidase activity were examined. The reslts suggest that cancer patients might have a "factor" or "factors" in their plasma which interfereu (s) with phagocytosis and which promote (s) exocytosis of lysosomal enzymes ("reverse endocytosis") from from autologous polymorphonuclear leucocytes as well as from cells obtained from normal healthy donors.


[Changes in several lipid metabolism indices following death and resuscitation]. The lipolytic activity in the adipose tissue, unesterified fatty acids (UFA) in the blood and adipose tissue, as well as ketone bodies and beta-lipoproteins in the blood were determined in dogs during dying of acute blood loss and the restorative period after the revival of the organism. During agony the activation of lipolysis in the adipose tissue, a decrease of UFA and beta-lipoproteins and an increase of ketane bodies contents in the blood were detected. At the end of the third minute of clinical death there occurred a depression of lipolysis and an increase of UFA content in the adipose tissue. One hour after the revival of the organism the blood UFA content and beta-proteins decrease, but the ketone bodies content rises; simultaneously there occurs some reduction of lipolytic activity of the adipose tissue. At the late postreanimation period (in 1, 3, and 7 days) an activation of lipolysis in the adipose tissue and an increase of UFA, ketone bodies, and beta-lipoproteins content in the blood was noted. The adipose tissue UFA content was low during the postreanimation period. The given results have shown that the changes in the lipid metabolism could play some role in the pathogenesis of non-reversibility during dying and after the revival of the organism.


[Role of corticosteroids in regulating splenic nucleic acid metabolism following severe mechanical injury]. The author studied the RNA and DNA concentration in the spleen of control rats, which had been adrenalectomized and had received ACTH before the trauma and 10 min, 5 and 10 hours after the beginning of the limbs' soft tissue crushing. The changes of nucleic acid metabolization in the spleen during the trauma were caused by the influence of corticosteroid hormones.


[Effect of thyrotropin releasing hormone thyroxine on the activity of cytochrome oxidase in rat adenohypophysis]. Thyrotropin releasing-hormone (TRH) increased the activity of cytrochrome C oxidase in a concentration of 0.01 and 1.0 microgram/ml in the adenohypophysis of rats fed methylthiouracil for 6 weeks. This effect of TRH on the activity of the enzyme was blocked with T4 added to the incubation medium in a concentration of 20 microgram/ml. Actinomycin D (20 MICrogram/ml) prevented the block of the enzyme with thyroxin. In a concentration of 0.01 microgram/ml TRH, and in a concentration of 2.0 microgram/ml T4 failed to change the activity of cytochrome oxidase in the adenohypophysis of normal and partially thyroidectomized rats.


[Organ and tissue cyclic adenosine-3', 5'-monophosphate during the process of the body's adaptation to extreme factors]. Various by nature extreme factors (hypoxic hypoxia, CO intoxication, chemical influence, noise and hypokinesia) caused similar changes in cAMP levels in different organs of albino rats (liver, the great cerebral hemispheres, heart). There was a significant increase in cAMP concentrations during the first stages of the influence and a progressive fall when the influence was continued. This fall was particularly expressed when the extreme factors used were of marked intensity. The authors consider the universality of the ascertained regularity to be a reflection of one of the central adaptive mechanisms of cell and the organism as a whole.


[Influence of single and repeated pregnancies on the incidence of primary tumors induced by SV40 virus in hamsters]. The influence of the single and repeated pregnancies on the SV40-induced carcinogenesis in Syrian hamsters was studied. It was shown that the frequency of the primary tumours in females which become pregnant 1-5 times during the latent period of SV40 carcinogenesis was significantly lower in comparison with virgin females observed in the same experiment. However, these differences apparently had no connection with the immunization of multiparous females with embryonic antigens, as the frequency and the time of appearance of primary tumours in males were the same as in multiparous females.


A conformational analysis for leucine-enkephalin using activity and binding data of synthetic analogues. 1. Leucine-enkephalin and some analogues were assayed for activity in vitro on the mouse vas deferens and for binding to opiate receptors from rat brain. 2. The experimental data were analysed in terms of the stringency for glycine, a D-amino acid or an L-amino acid at each position in the peptide. 3. The observed configurational specificity was compared with the stringency that would be predicted to occur if enkephalin adopted certain hydrogen-bonded conformations at the receptor. 4. A small subset of the conformations examined was found to be compatible with the experimental data.


Cannabis, catecholamines, rapid eye movement sleep and aggressive behaviour. 1. Previous work from our laboratory has shown that cannabis induces aggressive behaviour in rats that have been deprived of rapid eye movement (REM) sleep. It was suggested that this effect was related to brain catecholamines, with dopamine playing an agonist role and noradrenaline an inhibitory one. The present paper describes new experiments dealing with this subject. 2. Previous REM sleep-deprivation enhanced both delta9-tetrahydrocannabinol (THC)-induced hypothermia and nomifensine effects on aggressive behaviour. 3. A marihuana extract decreased brain dopamine turnover in REM sleep-deprived rats, an effect not observed in non-deprived rats. Noradrenaline metabolism was not altered. 4. Fighting behaviour was elicited in REM sleep-deprived rats treated with 4 different dopamine-beta-hydroxylase inhibitors. 5. Apomorphine, nomifensine and delta9-THC administered to non-deprived rats pretreated with bis(4-methyl-1-homopiperanzinyl-thiocarbonyl) disulphide (Fla-63), induced fighting behaviour. 6. Nomifensine and apomorphine induced fighting in non-deprived rats pretreated with delta9-THC. 7. Clonidine inhibited the fighting elicited in REM sleep-deprived rats by either delta9-THC or Fla-63 pretreatment. 8. The data are discussed in terms of the influence of REM sleep-deprivation (or the stress associated with deprivation) on the response to dopaminergic drugs and cannabis. Taken together they emphasize the participation of brain dopamine and noradrenaline systems in the aggressive behaviour studied.


Coronary reactions to cardiac hyperactivity and to hypoxia in isolated perfused heart of rat. 1. Continuous recording of cardiac force of contraction, heart rate and coronary flow from isolated perfused hearts of rats was used to study coronary reactions: (a) to cardiostimulation with noradrenaline, CaCl2, or electrically induced tachycardia; (b) to short duration stoppage of coronary inflow (hypoxia). 2. The heart rate was controlled by electrical pacing. Coronary vasodilatation resulted from cardiostimulation or hypoxia. This coronary response was greater at higher heart rates. 3. In parallel experiments administration of noradrenaline to hearts paced at different frequencies resulted in a rate-dependent elevation of adenosine-3',5'-cyclic monophosphate (cyclic AMP). 4. Duration of hypoxia leading to different degrees of reactive hyperaemia did not change the cardiac cyclic AMP levels. 5. Coronary vasodilatation due to increased cardiac metabolism produced by noradrenaline, Ca2+ or tachycardia was enhanced by the phosphodiesterase inhibitors diazoxide and papaverine while it was inhibited during the administration of prostaglandin E2.6. Reactive hyperaemia was unaffected by diazoxide, papaverine or prostaglandin E2. 7. Catecholamine depletion by reserpine did not influence metabolic coronary dilatation nor the reactive hyperaemic responses. 8. We postulate that there are at least two types of coronary reactions: one in response to hypoxia, 'reactive hyperaemia', and another resulting from cardiac hyperactivity, 'metabolic coronary dilatation'. The latter, blocked by prostaglandin E2 and enhanced by diazoxide or papaverine, would be triggered by cyclic AMP while reactive hyperaemia would result from other mechanisms.


Inhibition of plasma cyclic AMP, glucose and cortisol response to surgery by epidural analgesia. Cyclic AMP, glucose and cortisol in plasma were measured in three groups of patients undergoing hysterectomy. The operations were performed under general anaesthesia, under general anaesthesia combined with epidural analgesia and under epidural analgesia alone. Surgery elicited a significant rise in plasma cyclic AMP, glucose and cortisol when performed under general anaesthesia alone. Epidural analgesia extending from T4-6 to S5 combined with general anaesthesia abolished the rise in cyclic AMP and reduced the increase in glucose and cortisol and epidural analgesia alone extending from T4 to S5 blocked the rise in glucose and cortisol as well as that in cyclic AMP. The results support the theory that afferent nerve impulses from the area of trauma are of major importance for the catabolic state induced by surgical procedures and indicate that anaesthetic management which includes blockade of afferent nerve impulses which includes blockade of afferent nerve impulses from the area of trauma can be reduce the catabolic response to surgery. These observations could be of value in the operative management of patients with diabetes mellitus and possibly in other groups by patients with a high surgical morbidity.


Reversible blockade of PGO waves and concomitant modifications of thalamic unit activity in chronic cats. The reversible blockade of geniculate PGO waves was carried out by localized cooling of the ponto-geniculate pathways during paradoxical sleep in chronic cats. In parallel, long-term recordings of unit activity were performed in non-specific visuooculomotor thalamic structures where no PGO waves occurred. Blockade of geniculate PGO waves is concomitant with a preferential modification of the phasic discharge of the unit activity (i.e. time-locked with rapid eye movements) when compared to the tonic discharge (i.e. without rapid eye movements). These results suggest that pontine generators of PGO waves could act upon every neuron of the central nervous system. The significance of such an extensive ascending system, the blockade of which does not interrupt paradoxical sleep episodes, still remains puzzling.


Characterization of a myelin-related fraction (SN 4) isolated from rat forebrain at two developmental stages. A myelin-related fraction (SN 4) was isolated from forebrain of 17- and 40-day-old rats. Fraction SN 4 was obtained as a supernatant in a slow speed differential centrifugation of a myelin fraction. In contrast to multilamellar myelin fraction, SN 4 consisted of small vesicular profiles of a mixture of single membranes and some triple-layered structures. All typical myelin components were found in the SN 4 fraction from adult rat brain but their relative proportion was different from that of myelin: Wolfgram protein, myelin glycoproteins and 2',3'-cyclic nucleotide 3'-phosphohydrolase were increased, while basic proteins and proteolipid protein were decreased significantly. In contrast, the lipid composition appeared very similar to the one found in myelin. SN 4 from 17-day-old rat brains was essentially similar to that from adults, except that the major myelin glycoprotein was not enriched in comparison to myelin. Developmental changes found in myelin were also present in the SN 4 fraction. The specific radioactivity of the fucose-labeled major myelin glycoprotein was similar in SN 4 and in myelin. The particular composition of fraction SN 4 suggests that this material is not significantly contaminated by non-myelin-related membranes but rather supports the hypothesis that it could be enriched in a membrane representing a zone of transition during the formation of myelin and which is subjected to a remodelling of its protein components.


Presynaptic origin of penicillin after discharges at mammalian nerve terminals. The site of origin and mechanism underlying the generation of repetitive after-discharges produced by penicillin was studied in the isolated rat phrenic nerve-hemidiaphragm preparation. Application of low concentrations of sodium penicillin to the bathing solution initiated bursts of antidromic action potentials originating at or near the motor nerve terminals following single orthodromic stimuli to the nerve. Afterdischarges could not be elicited by direct stimulation of the muscle fibers alone, or when the nerve trunk was isolated from the neuromuscular junction and exposed to penicillin. D-Tubocurarine applied in doses sufficient to abolish postsynaptic responses did not diminish penicillin-induced after discharges. At concentrations which most reliably produced repetitive firing (5000 IU/ml; 8.5 mM), penicillin did not accelerate the frequency of spontaneous transmitter release (MEPPs), yet significantly increased the relative excitability of nerve endings to extracellular stimulation. It is concluded that penicillin acts directly and preferentially on presynaptic nerve terminals to induce repetitive afterdischarges which arise independently of postsynaptic depolarization, transmitter-mediated potassium efflux, or muscle fiber contraction. The results suggest that the convulsant effects of penicillin at a mammalian neuromuscular junction are due to non-depolarizing alterations in the intrinsic excitability of the terminal membrane which increase the probability of suprathreshold depolarizations during the recovery period of spike electrogenesis. Several models of the mechanisms which might produce hyperexcitability at presynaptic nerve terminals are discussed.


[Cells with phosphatase activity in the walls of subcutaneous capillaries from normal and inflamed rat tissue]. In the wall of rat's capillary blood vessels the adenosine triphosphatase is located on the endothelium, the 5'nucleotidase on the perithelial cells. The perithelial cells activities are strong during the inflammatory state, but at the beginning of the phenomenon the nucleotidasic cells become free into the oedematous tissue by preceding and accompagning the fibroblastic differenciation cells.


[Experimental nephrotic syndrome in the rat. Biologic parameters and study of several hydrolases in different purified kidney fractions]. The induction of nephrotoxic nephritis in rats with rabbit antibodies preparation results in proteinuria, hypoproteinemia and hyperlipidemia with little glomerular lesions. A study of some hydrolases in cortex and medulla on one hand and glomerular and tubules on the other, showed changes in the activities of following enzymes. 1) A 20-30 % decrease in Na+, K+ dependent ATP-ase in whole kidney. 2) A 20 % decrease in beta-galactosidase activity in glomerular and medulla. 3) A 20 % increase of arylsulphatase A activity in tubules. These results are discussed in the light of the present knowledge of sulphatide metabolism in kidney.


[Effect of NADH and several Krebs cycle substrates on the endogenous metabolism of Pseudomonas fluorescens (type S)]. Simultaneous addition of succinate (or fumarate) and NADH to a non-proliferating suspension of P. fluorescens results in an enhancement of the oxydation of the endogenous substrates. This conclusion is in accord with the determination of the respiratory and the oxidation quotients of the analysed exogenous substrates.


[Reduction of adrenaline-dependent lipolysis in the adipose tissue of adrenalectomized dogs by an alpha adrenergic effect]. In dog, surrenalectomy reduces isoprenalino-depending lipolysis but still more adrenalino-depending lipolysis. Phentolamine potentializes the response to adrenaline but does not that observed with isoprenaline. Surrenalectomy unmasks an alpha adrenergical effect of adrenaline on adipocytes of dog adipose tissue.


Effect of the inhibitor of angiotensin I converting enzyme on endocrine function and renal perfusion in haemorrhagic shock. The present study was undertaken to investigate the effect of inhibition of angiotensin I converting enzyme on endocrine responses and renal perfusion in haemorrhagic shock. Plasma levels of vasopressin (VP), aldosterone, and plasma renin activity (PRA), and renal cortical tissue blood flow were measured both in control dogs and in those treated with an inhibitor of angiotensin I converting enzyme (SQ20881). The drug was administered with an intention of preventing vasoconstriction induced by angiotensin II and to improve tissue perfusion. No significant differences were found in plasma levels of vasopressin, aldosterone and plasma renin activity between control and SQ20881-treated groups. Secondary elevation of blood pressure following haemorrhage was significantly delayed and reduced by the administration of the inhibitor. Decrease in renal cortical tissue blood flow was observed in the SQ20881-treated group. It is suggested that angiotension II appears to play a role in spontaneous recovery of arterial blood pressure following haemorrhage. Furthermore, angiotensin II does not seem to play an important role in the stimulation of secretion of vasopressin in response to haemorrhage. Our data failed to demonstrate any favourable effects of inhibition of angiotension I converting enzyme on renal perfusion during haemorrhagic shock in dogs.


Avoidance of vascular complications associated with the use of dopamine. Dopamine, dihydroxyphenylethylamine has three distinct actions depending on dosage. Low doses in the range of 1-2 mcg/kg/min result in vasodilatation. Medium doses of 2-10 mcg/kg/min increase cardiac output due to beta adrenergic action, while above 10 mcg/kg/min a potent vasoconstrictor effect predominates. Because of this last action great care must be used to avoid extravasation of infusions of dopamine directly into the tissue. Dopamine is a very useful agent but carries great potential to do local damage. Administration through a catheter placed into a large vein is recommended. Should extravasation occur, prompt infiltration of the area with phentolamine and a local anaesthetic, local cooling and regional sympathetic block may reduce the damage. The case is reported of a patient who suffered extensive necrosis in the forearm following local extravasation of dopamine.


Thyroid and breast cancer following childhood radiation. A 35-year-old man who received radiation for thymic enlargement as a child and subsequently developed both thyroid and breast cancer is reported. This appears to be the first case in which this association has been reported in a male.


Immunotherapy of a guinea pig hepatoma with living BCG: a frozen liquid and a lyophilized preparation are equally active. A freeze dried preparation of BCG with a low ratio of living to dead organisms (LV) was compared to a frozen liquid preparation with high viability (HV) for its ability to eradicate established dermal tumors and microscopic lymph node metastases in guinea pigs. The cure rate achieved by the intralesional injection of LV-BCG did not differ significantly from that of HV-bcg when similar numbers of viable organisms were injected.


Establishment of an Epstein-Barr virus-negative B-cell lymphoma line from a Japanese Burkitt's lymphoma and its serial passage in hamsters. An Epstein-Barr virus (EBV)-negative lymphoma line (JBL) was established in vitro from pleural effusion of an EBV-seropositive 29-year-old Japanese female with Burkitt's lymphoma. JBL cells as well as her original lymphoma cells bore monoclonal surface IgM with lambda light chains. The JBL line grew in single cell suspension with a doubling time of 30 hours. Attempts were made to serially transplant JBL cells in antilymphocyte serum-treated newborn hamsters; intraperitoneal implantation of 1-3 X 10(7) cells gave rise to invasive tumors in all recipients with death after 10 to 14 days. The hamster-passage line, now in the 9th passage, has been converted to an ascitic form with progression to leukemia in some animals. A "starry sky" pattern closely resembling the human tumor material was preserved in every tumor through serial animal passage.


Malignant myoepithelioma of the parotid gland. Myoepithelial cells are a significant component of most types of salivary gland neoplasms. A small but increasing number of case reports have also shown that pure myoepitheliomas form a distinct class of neoplasms with unique histological features. Previously reported cases have been either benign or, at most, locally aggressive. We present here a case of a malignant myoepithelioma of the parotid gland, confirmed by electron microscopy, that was locally aggressive and eventually metastasized. Review of this case and of existing reports indicates that myoepitheliomas exhibit a wide range of biological behavior that seems to correlate with their histological appearance.


Extrarenal Wilms' tumor: report of a case and review of the literature. A case of an extrarenal Wilms tumor arising in the inguinal region of a 2-month-old boy is described. A review of the literature indicates that nine well-documented cases of extrarenal Wilms tumor have previously been described. Four of these tumors arose within teratomas. The remaining five cases, in which no teratomatous components were encountered, probably arose in embryologic rests of renal tissue.


Multiple childhood osteosarcomas in an American Indian family with erythroid macrocytosis and skeletal anomalies. Three of nine children of possibly consanguineous American Indian parents developed typical osteosarcoma in a 2-year period. Etiologic investigations detected limb anomalies and elevated mean corpuscular volumes (98--109 micrometer3) in the surviving tumor patient, several of her sibs, and her father. Limb anomalies included simple clinodactyly with brachymesophalangy, absence of one digital ray of the foot, and bilateral radioulnar synostosis. The red cell macrocytosis was not accompanied by anemia or explained by the usual causes. No unusual environmental exposures were found and screening for possible oncogenic viruses by culture, electron microscopy, and serology was negative. All family members had elevated antibody titers to Epstein-Barr viral antigens. The proband and her father had excessive chromosomal breaks in the bone marrow. This unusual familial pattern of osseous malignancy and malformation and defective erythropoiesis, tentatively called OSLAM syndrome, may represent impaired regulation of bone development.


Central nervous system tumors in children. Of 488 central nervous system neoplasms occurring in children over a 39-year period, 467 were intracranial and 21 were intraspinal. The most common intracranial neoplasms were astrocytoma (28%), medulloblastoma (25%), ependymal neoplasm (9%), craniopharyngioma (9%), and glioblastoma multiforme (9%). The median age at diagnosis was 6 years with a male-to-female ratio of 1.3:1. Overall mean survival was 53.4 months and varied greatly relative to the type of tumor and the location. Of the intraspinal neoplasms the most frequently noted were the astrocytoma (47%) and the ependymal neoplasma (24%). The median age at diagnosis was 10 years with a male-to-female ratio of 1:1. The average survival from diagnosis (54.1 months) was comparable to that of intracranial neoplasms. Detailed analyses of each histological type of tumor relative to age at diagnosis, sex, anatomical location and survival from diagnosis are reported for both intracranial and intraspinal neoplasms.


Wilms' tumor in New York State: epidemiology and survivorship. The outcomes during the period 1950--1972 were compared for Wilms' tumor patients in Erie County, New York (Buffalo and environs) and in a random selection of 23 counties having much smaller populations. For the Erie cohorts of 1967 to 1972 an 87 per cent 7-year survival rate was found as compared with a 50 per cent survival for the corresponding cohorts of the less populous couties. For the years 1960--1966 the 5-year survival rates were respectively 67 and 25 per cent and for the decade 1950--1959, 26 and 23 per cent. The principal conclusion is that within the last 15 years the Erie residents have fared better than residents of the smaller counties. The difference is attributed to the better treatment and care available at some of the hospitals in Buffalo. Data on incidence, age at diagnosis, male/female ratio, and laterality are presented.


Changes in polyamine levels and protein synthesis rate during rat liver carcinogenesis induced by 4-dimethylaminoazobenzene. The concentrations of putrescine, spermidine, and spermine in liver of rats fed on 4-dimethylaminoazobenzene and in the resultant hepatomas were found to be significantly higher than were those observed in normal liver from rats of the same strain, sex, and age. These modifications were due to the carcinogen and not to the special low-riboflavin diet used to obtain the carcinogenic effect of 4-dimethylaminoazobenzene. The first change observed during liver carcinogenesis was the early increase in the putrescine level, followed by an increase of spermidine and spermine, which reached maximum levels in growing hepatomas. A significant increase of urinary polyamines was also observed in tumor-bearing rats. Experiments on leucine incorporation into proteins of tissue slices, which were obtained from the same tissues on which polyamine determinations were carried out, showed that in rat liver carcinogenesis the rate of protein synthesis was well correlated with the polyamine levels. These results suggest that polyamines may play a role in the process of carcinogenesis and in tumor protein synthesis in vivo.


Initial reaction of 3-ethyoxy-2-oxobutyraldehyde bis(thiosemicarbazonato) copper(II) with Ehrlich ascites tumor cells. The antineoplastic agent, 3-ethoxy-2-oxobutyraldehyde bis(thiosemicarbazonato) copper(II), undergoes a pseudo-first-order decomposition in the presence of viable Ehrlich ascites tumor cells. Physical and chemical methods show that the copper is reduced in this process, that thiols are involved in the reaction, and that the thiosemicarbazone ligand is displaced from the copper ion. The widespread distribution of the cell-bound copper suggests several modes of cytotoxicity.


Mutagenicity of diallate, sulfallate, and triallate and relationship between structure and mutagenic effects of carbamates used widely in agriculture. In an investigation of the mutagenic properties of 20 carbamate herbicides and fungicides by use of the Salmonella/microsome mutagenicity test as developed by Ames et al. (Mutation Res., 31: 347-364, 1975), we have found that three thiocarbamate compounds, diallate, sulfallate and triallate, are mutagenic in the presence of a liver microsomal fraction on strains TA1535 and TA100. This indicates that the metabolic products of these thiocarbamates are causing base-pair substitutions. Since the 2-chloro-allyl group is common to the three mutagenic compounds but is not common to the 17 nonmutagenic compounds, a metabolic derivative of this group is probably responsible for the mutagenic activity.


Metabolic control of glycolysis in normal and tumor permeabilized cells. Our previous reports have presented evidence suggesting the existence in tumor cells of a second control site of glycolysis of pyruvate kinase as a competition for adenosine diphosphate between this enzyme and mitochondria, which is responsible for the Crabtree effect. Now, by using cells partially permeabilized to nucleotides and phosphorylated substrates, we provide evidence supporting the existence in hepatocytes of a partial control by adenosine triphosphate at phosphofructokinase, which is followed by the total control by adenosine triphosphate at pyruvate kinase. The partial or nonoperation of this second site in Ehrlich ascites tumor cells appears to be the cause for the characteristic aerobic glycolysis, Crabtree effect, and low Pasteur effect of these cells.


Ultrastructure of the thyroid gland in goitered coho salmon (Oncorhynchus kisutch). Thyroid tissues in coho salmon (Oncorhynchus kisutch) goiters showed marked regional variation in structure and ultrastructure. For the most part the follicles were small, were composed of tall epithelial cells, and contained little or no colloid material. The follicle epithelial cells were essentially similar in fine structure to hyperactive thyroids in mammals and one type of follicle in congenital goiters in humans. A second type of follicle, present in human goiters, was not found in coho salmon. The follicle cells in coho salmon contained dense bodies similar to lysosomes in other species, together with larger organelles which could be colloid droplets. There was also an extensive system of dilated rough endoplasmic reticulum and large areas of Golgi membranes. A marked feature was the presence of homogeneous belectron-dense material in the intercellular spaces between adjacent follicle cells.


Location of adult and fetal aldolases A, B, and C by immunoperoxidase technique in LF fast-growing rat hepatomas. The resurgence of aldolase isozymes in cancerous tissues is a well-known but poorly understood phenomenon. This resurgence poses the problem of whether or not adult and fetal aldolase isozymes are produced by the same cells. For clarification of this question, the immunoperoxidase technique was used to locate aldolases A, B, and C in one type of fast-growing hepatoma, the LF hepatoma and, by comparison, in normal adult liver. Under optical microscopy, aldolases A and C were located in the cytoplasm of almost all of the cancerous cells. An isozyme antigenically identical with aldolase B was also demonstrated to be present in almost all of the cells, but the reaction indicating the presence of this isozyme was weaker. In normal adult liver, only aldolases A and B were demonstrated to be present in almost all the hepatocytes. Under electron microscopy in LF hepatoma, the three isozymes were found to be present mainly in the cytoplasm. These facts suggest that the three types of aldolase are very probably present in the same cells at the same time, and they provide indirect arguments leading us to think that the resurgence of fetal aldolase isozymes in cancer is not the consequence of cellular selection but is due to a disturbance at the gene control level.


Patterns of spontaneous metastasis of transplantable hepatocellular carcinomas. A number of transplantable rat hepatocellular carcinomas of varied phenotype were examined for their ability to metastasize. A striking diversity of pattern was observed, ranging from presentation in almost every organ to none at all. No relationship between metastatic capability and growth rate, tumor size, chromosome composition, or other functional characteristics was noted. Interestingly, several of the tumors demonstrated a lymphatic to vascular route of spread, very similar to that of human tumors.


Turnover of cellular carbohydrates in normal and Rous sarcoma virus-transformed cells. We have analyzed the distribution of glucosamine-labeled polymers on the cell surface, in the growth medium, and inside the cell and the net turnover of these polymers during the process of malignant transformation of chick embryo fibroblasts by Rous sarcoma virus. The distribution of label and the turnover kinetics for hyaluronic acid, total glycoproteins, and chondroitins were found to be identical in both normal and transforming cultures. We conclude that nonspecific degradative processes are probably not involved in causing transformation-related alterations in cell surface carbohydrates, althougn degradation of specific macromolecules is not excluded.


Immunotherapy of established micrometastases with Bacillus Calmette-Guérin tumor cell vaccine. We evaluated the use of Bacillus Calmette-Guérin admixed with tumor cells as a vaccine to induce systemic tumor immunity for therapy of subclinical (micrometastatic) disease. In several experiments inbred strain 2 guinea pigs were given i.v. injections of either 10(4), 10(5), or 10(6) syngeneic L10 hepatocarcinoma cells, and initial vaccinations were administered either 1 or 4 days after tumor inocluation. Variables in vaccine preparation, such as ratio of viable Bacillus Calmette-Guérin organisms to tumor cells, procedures for freezing the tumor cells, X-ray treatment of tumor cells, and vaccination regimen were evaluated. The studies demonstrated that under defined conditions nontumorigenic vaccines of Bacillus Calmette-Guérin and tumor cells can cure the majority of animals of otherwise lethal visceral micrometastases.


Protection against MOPC-315 plasmacytoma by immunization with C-type particle preparations. Mice were given injections of C-type particles extracted from the ascitic fluid of plasmacytoma-bearing mice. These particles, extracted from MOPC-315 tumor-bearing mice and injected into BALB/c mice, protected them against challenge with MOPC-315 tumor cells. The protection was dependent upon tumor cell dose; 66% survival was observed with a lethal dose of tumor cells. No protection was observed against challenge with another plasmacytoma (S13). Attempts to protect mice against S-13 plasmacytoma by immunizing them with C-type particles originating from S-13 tumor-bearing mice were unsuccessful.


Inhibitory effect of hypothalamic lesions on liver tumor induction by N-2-fluorenylacetamide in male rats. Bilateral electrolytic lesions were placed in the median eminence area of the hypothalamus in 12-week-old male Wistar rats. Sham-operated and untreated control rats were also included. Two weeks later, one-half of them were given 0.03% N-2-fluorenylacetamide incorporated into the diet for 16 weeks with adequate resting periods in between. The animals were killed 34 weeks after the last carcinogen feeding. The results show that lesions in the hypothalamus effectively inhibited liver tumor formation (0 of 16, 0%). In contrast, the incidence of hepatocellular carcinomas in sham-operated rats was 38.5% (5 of 13), and that of untreated controls was 42.9% (6 oactive thyroid glands, and shorter nasoanal lengths were observed in rats with lesions in the hypothalamus irrespective of carcinogen treatment. It is apparent from these data that lesions in the median eminence area of the hypothalamus inhibit the induction of liver carcinogenesis with N-2-fluorenylacetamide in male rats.


General immunocompetence of rats bearing avian sarcoma virus-induced intracranial tumors. The mitogenic responsiveness of spleen cells obtained from avian sarcoma virus-inoculated Fischer 344 rats was studied. Sixty % of the rats had astrocytomas, 13% had sarcomas, 7% had mixed gliosarcomas, and 20% had no evidence of tumors. Only spleen cells from rats bearing astrocytomas had significantly diminished responses to phytohemagglutinin and concanavalin A (Con A) when compared to control responses. The decreased responsiveness observed with phytohemagglutinin was limited to the optimal concentration range (10 and 20 microgram) while a broader concentration of Con A (0.01 to 50 microgram) induced significant suppression. Moreover, a more profound immunosuppression was observed with Con A. The results also demonstrated that spleen cells from rats with the largest astrocytomas exhibited the greatest suppression. From the results of this study, it appears the avian sarcoma virus-induced astrocytoma in rats is an immunological parallel of the human disease based on the loss of general immunological competence as assessed by responsiveness of lymphocytes to phytohemagglutinin and Con A.


Localization of meso-tetra(p-sulfophenyl)porphine in murine sarcoma virus-induced tumor-bearing mice. meso-Tetra(p-sulfophenyl)porphine (TPPS4) has been found to accumulate in the tumors of mice bearing a murine sarcoma virus-induced rhabdomyosarcoma to a greater degree than in all other tissues except for the kidney. Tumor to tissue ratios of from 3:1 (tumor to liver) to 9:1 (tumor to muscle) were observed. The uptake of TPPS4 was found to be dose dependent, and the highest tumor to tissue ratios were found 96 hours after injection. The water-soluble TPPS4 appears to be cleared through the kidneys and absolute concentrations there are reduced with increasing time after injection. These data are compared with previous studies on TPPS4 localization in Walker 256 carcinosarcoma-bearing mice. In view of this localizing ability and its relative lack of toxicity, TPPS4 seems to represent an ideal candidate for a tumor-localizing agent if high levels of the porphyrin in the kidneys can be reversed.


Scintigraphic visualization of myocardial infarcts in baboons using thallium-201 and  technetium-99m pyrophosphate. 4 baboons with myocardial infarcts were evaluated using thallium-201 for myocardial imaging and 99Tcm pyrophosphate for infarct visualization. Scintiphotographic findings were compared with the size of myocardial infarcts calculated from measurements of the activity of MB isoenzymes of creatine kinase (CK-MB) in serum and in the myocardium at autopsy, as described by Sobel's method. Lack of thallium-201 accumulation was noted in left ventricular infarcts of 3 of the 4 baboons. These same areas localized 99Tcm pyrophosphate administered 24--30 h after infarction.


The effect of diet on the ultrastructure of the mid-gut cells of Nasonia vitripennis (Walk.) (Insecta: Hymenoptera) at various ages. The ultrastructure of mid-gut cells of female Nasonia fed on a diet of 10% sterile sucrose is described. There are extensive alterations in cell organelles, particularly the mitochondria, rough endoplasmic reticulum (R.E.R.) and lipid inclusions, when compared to similar insects fed a normal diet of dipteran pupae. A proportion of the mitochondria found in the apical cell region are enlarged in size and contain electron-dense granules. The remaining mitochondria are smaller, but also contain electron-dense granules. Cytochrome-c oxidase activity appears to be absent from the enlarged mitochondria. The R.E.R. appears reduced in many cells of the 1 day old, sugar-fed insects, however, this component fluctuates throughout the remaining life span. The lipid inclusions prominent in the 1 day old, pupae-fed insects are not present in sucrose-fed females of the same age, but lipid deposition was recorded later in the life span. There are many large residual bodies and cytolysosomes present in the old, sugar-fed insects. These changes in ultrastructure are discussed in relation to diet and longevity.


Intramitochondrial bodies in bovine adrenocortical cells. Electron dense, homogeneous, mostly round intramitochondrial bodies were found in bovine adrenal glands, predominantly in the zona glomerulosa. The histochemical results obtained suggest that these bodies might contain a considerable amount of protein, although the possibility that substances other than proteins are contained in these bodies cannot be completely ruled out. The physiological significance of the intramitochondrial bodies remains unclear.


The mechanisms of hepatic and renal metallothionein biosynthesis in cadmium-exposed rats. The mechanisms by which cadmium (Cd) stimulates the synthesis of hepatic and renal metallothionein were investigated in rats. In the liver the incorporation of 35S and 14C labels from cystine showed a lag period of about 2 h and maximum incorporation into metallothionein took place 8--12 h after injection of 30 mumol CdCl2/kg. The incorporation of the labels into renal metallothionein was faster than into the hepatic protein; the lag period was less than 1 h and maximum incorporation occurred 4--5 h after the Cd injection. Cycloheximide inhibited the amino acid incorporation into both hepatic and renal metallothionein. Actinomycin D treatment, however, inhibited hepatic metallothionein synthesis only. These results suggested that at the dose level employed Cd regulated the biosynthesis of metallothionein in the liver at the transcriptional level and in the kidney at the translational level.


Poxvirus in West African nonhuman primates: serological survey results. Ten species of nonhuman primates in West African habitat were analysed for variolavaccinia subgroup haemagglutination-inhibition (HI) and neutralization antibodies. The animals were taken in 27 different sampling zones in parts of the Ivory Coast, Mali, and Upper Volta. Of the 195 tested, 15 (8%) had elevated HI antibodies after nonspecific reactions were reduced with potassium periodate pretreatment. Positive neutralization antibodies were found in 21% (44 of 206). Antibodies were detected in serum from monkeys living near two areas where monkeypox cases in humans had occurred. Four samples were tested for monkeypox specific antibodies using an indirect immunofluorescent test; 3 were positive. Despite the prevalence of poxvirus antibodies in monkeys (and other animals) in West Africa, smallpox eradication has been maintained in the area since 1970; thus, animal reservoirs of poxvirus appear to pose no threat to the worldwide smallpox eradication programme.


Differentiation of variola, monkeypox, and vaccinia antisera by radioimmunoassay. Poxvirus antisera adsorbed with "homologous" and "heterologous" poxvirus-infected chorioallantoic membranes (CAM) were differentiated by solid-phase radioimmunoassay (RIA). Mixtures of the antiserum dilutions and infected CAM were added directly (without centrifugation) to poxvirus-infected CAM antigens affixed to wells of microtitration plates. The affixed antigens combined with unadsorbed antibodies, and the cross-reactive antigen-antibody complexes were removed by washing. The results showed that adsorption of an antiserum with variola-, vaccinia-, or uninfected-CAM antigen and subsequent reaction of each in RIA with monkeypox- and uninfected-CAM antigens allowed the identification of antivariola, antivaccinia, or antimonkeypox sera.


The viral etiology of acute respiratory infections in children in Uganda. The role of viruses in respiratory diseases of young children in Uganda was studied. A viral etiology was established in 36% of the infections investigated. The most important pathogens were found to be respiratory syncytial virus and parainfluenza viruses, which were responsible for 26% of infections investigated. They caused both upper and lower respiratory tract diseases. There was little or no seasonal variation in the etiology of these infections. Adenoviruses were found to be less important and were etiologically related to only 4% of respiratory disease cases. Influenza viruses and enteroviruses were also found to be associated with respiratory infections. However, they were less frequent and their role was insignificant. The role of multiple virus infections was also insignificant.


Buffer-exchange column for rapid separation of carcinoembryonic antigen from perchloric acid. A Sephadex G-50 medium (Pharmacia) buffer-exchange column has been developed for rapidly changing the medium for carcinoembryonic antigen from perchloric acid to acetate buffer. Analytical recovery of the extracted antigen exceeded 95%. Plasma for analysis can be so prepared and samples ready for analysis within an hour. The technique also results in uniform assay volume, thus eliminating this variable from the assay.


Lipoprotein cholesterol in the serum of children, as determined independently by two different methods. We separately evaluated split specimens of serum for lipoprotein cholesterol in 38 five- to 14-year-old children, by the heparin/Ca2+ precipitation method and the ultracentrifugation/dextran sulfate precipitation method. Statistical analysis of the results indicated an excellent agreement (especially in beta- and alpha-lipoprotein cholesterol values) between results by the two methods. The percentage of alpha-lipoprotein cholesterol (mean +/- SD: 38.6 +/- 5.0) in children further confirmed the earlier observations that cholesterol derived from alpha-lipoprotein in children constitutes a relatively greater part of the serum total cholesterol than is true of adults.


Use of the Du Pont aca to measure cholesterol in high-density lipoprotein fractions prepared by the heparin/Mn2+ precipitation method. The enzymatic cholesterol method used with the Du Pont aca has been modified to provide a reliable measurement of high-density lipoprotein cholesterol in serum after heparin/Mn2+ precipitation of the low- and very-low-density lipoproteins. Interference by Mn2+, equivalent to about 90 mg of cholesterol per liter, is decreased to less than 40 mg of cholesterol per liter by the presence of ethylenediaminetetraacetate (8 mmol/liter) in the diluent; the residual effect of Mn2+ is compensated by calibrating the aca with standards containing Mn2+ and heparin. With an 80-microliter sample, the sensitivity is 236 muA/mg per liter and linearity ranges from 50 to 1500 mg/liter. Average analytical recovery of cholesterol added to the high-density lipoprotein fraction was 103%. Diluted fractions give the expected results. Between-run reproducibility (CV) is 1.3 and 1.6% at 463 and 554 mg/liter. Correlation with the Lipid Research Clinics' procedure (25 samples) gave a regression line of y(aca) = 1.039x- 15, and a correlation coefficient of 0.997.


Serum and lipoprotein apolipoprotein B levels in normal subjects and patients with hyperlipoproteinaemia. Apolipoprotein B (apo B) levels were measured by radioimmunoassay in the serum and lipoproteins from normal subjects and patients with hyperlipoproteinaemia. The total serum apo B concentration in normal subjects was 0.91 +/- 0.16 g/l (mean +/- S.D.); in type IIa hyperlipoproteinaemia it was 2.24 +/- 0.61 g/l; in type IIb, 3.05 +/- 1.24 g/l; in type IV, 2.24 +/- 0.99 g/l; and in type V, 1.30 +/- 0.16 g/l. In normal subjects 5.6 +/- 2.1% (mean +/- S.D.) of total apo B was present in very low density lipoproteins (VLDL) and 93 +/- 9% in low density lipoproteins (LDL). Corresponding values for type IIa were 3.8 +/- 1.9% and 93 +/- 3%, for type IIb, 9.9 +/- 7.5% and 91 +/- 1%, for type IV, 16.9 +/- 9.5% and 81 +/- 9%, and for type V, 38.4 +/- 11.0% and 52 +/- 8%. The ratio of cholesterol to apo B in serum was decreased in types IIa, IIb and IV, and increased in type V whereas the ratio of triglyceride to apo B in serum was decreased in type IIa, normal in type IIb and increased in types IV and V. The ratio of cholesterol to apo B in VLDL was increased in types IIa, IIb and V, but normal in type IV, whereas in LDL, this ratio was normal in types IIa and V but reduced in types IIb and IV. The ratio of triglycerides to apo B in VLDL was normal in types IIa, IIb and IV but raised in type V. In LDL, this ratio was increased in types IIb and IV but normal in types IIa and V.


Collagenase activity of rat kidney with immune complex glomerulonephritis. By means of a biological assay the collagenolytic activity of kidneys from rats with experimental immune complex glomerulonephritis and control animals was tested. We detected collagenolysis in both panels but found quantitative differences in the collagenase activity. The enzymatic collagenolysis was significantly increased (P less than 0.001) in the nephritic kidneys. From the fact that the chelating agent EDTA inhibited potentially the released enzyme it can be concluded that the origin is the polymorphonuclear granules.


Changes in rat serum lipoproteins following ligation of the bile duct. Lipoproteins in the serum of normal and of cholestatic rats have been studied by crossed immunoelectrophoresis coupled with separations by rate and isopycnic density gradient centrifugation and by gel exclusion chromatography. Normal rat serum contained distinct lipoprotein species closely analogous to human VLDL, LDL, HDL2 and HDL3. Three days after ligation of the common bile duct, there were major changes in the lipoproteins of rat serum. The amounts of VLDL, LDL and of a minor HDL component were elevated and several novel types of lipoprotein were detected. Three of these could be identified as the characteristic lipoprotein of cholestasis, LP-X, as an enlarged and modified HDL and as a lipoprotein of density 1.055 g/ml intermediate in size between LDL and VLDL. Some unusually small VLDL particles were also detected. It is concluded that the changes in rat serum lipoproteins following ligation of the common bile duct are very similar to the changes observed in cholestatic disease in human patients.


The mechanism of action of polypeptide hormones with special reference to insulin's action on glucose transport. In the last decade substantial progress has been made in elucidation of the mechanisms of action of polypeptide hormones. For instance, it has become clear that these hormones bind to their target cells and that at least one component of the binding is saturable. This phenomenon, which is usually called receptor binding, is believed to be the first and necessary event in the chain of processes which lead to effects of the hormone on their target cells. Receptor binding is just one aspect of the mechanism of action of polypeptide hormones. The properties of binding differ from hormone to hormone as do the events after binding. The purpose of this communication is to describe some aspects of the mechanism of insulin action on transmembrane glucose transport, and to disucss general aspects of polypeptide hormone action in relation to the specific findings with insulin. Comprehensive reviews on polypeptide hormone receptors have been published recently (Freychet, 1976; Kahn, 1976).


Prolactin secretion in girls with isolated gonadotrophin deficiency. Plasma prolactin, basal levels and the response to an i.v. injection of TRH (100 microgram/m2) was determined in five girls with isolated gonadotrophin deficiency of hypothalamic origin before and after at least 3 months cyclic replacement therapy with conjugated oestrogens (1.25 mg/day). The basal plasma prolactin levels were similar during both tests, however, during oestrogen therapy the mean peak response to TRH almost doubled and the sum of all the values obtained (basal, +15, +30 and +60 min) was significantly higher. It is of note that even upon prolonged oestrogen deprivation the releasable prolactin response to TRH was adequate.


E-rosette inhibiting substance in Hodgkin's disease spleen extracts. Patients with Hodgkin's disease often manifest impairment of cell-mediated immune responses in both in vivo and in vitro tests, as well as a markedly decreased percentage of E rosette-forming (T) lymphocytes in the peripheral blood. This report describes the inhibition of E-rosette formation by normal peripheral blood lymphocytes after incubation with extracts prepared from the spleens of patients with Hodgkin's disease. Such extracts also depressed E-rosette formation by the peripheral blood lymphocytes of patients with Hodgkin's disease after those cells had been restored to normal function by prior incubation in foetal calf serum. Similarly prepared extracts from the spleens of normal donors had no immunodepressive effect. The E-rosette inhibitory substance in the Hodgkin's disease spleen extracts was found to be a complex containing beta-lipoprotein, C-reactive protein, and C1q.


Transfer factor therapy in ataxia--telangiectasia. The effects of weekly doses of transfer factor in four patients with ataxia--telangiectasia were investigated following a total course of 2 months therapy. Transfer factor administration showed no influence on the absolute lymphocyte counts, T-cell rosettes or antibody titres to EBV, but it caused conversion of skin-test reactivity and production of MIF to various antigens. There was a dissociation in blastic transformation response, the skin-test responses and MIF production. Serum interferon levels were low before, and 2, 6 and 24 hr after, therapy. Clinically no improvement in infections was observed following transfer factor therapy.


Echo-encephalographic measurements (A-scan) of the lateral ventricles in children; its reliability. A positioning of the probe on the skull has been developed to increase the diagnostic accuracy of hydrocephalus. Echo-encephalography in the trigonum region of the ventricles is usually possible in children up to 13 years of age. In this region sources of error as the medial wall of the temporal lobe or the lateral wall of the temporal horn are eliminated. In hydrocephalic ventricles the ultrasound measurements in this region corroborate very well with the measurements found in the pneumo-encephalograms. This in contrast to the measurements found with the probe placed in the more conventional region. As the largest diameter of the skull, corresponds with the trigonum region, a very practical topographic mark for a reliable standardized method has been found. With more confidence the ventricle indices of 93 normal children, in the age of 1 to 13 years, were measured.


Computed tomography of supratentorial meningioma. The appearance on C.T. of 28 meningiomas is described. On the plain scan they presented as a rounded, well defined lesion with a high density in 18 cases and a density equal to or lower than brain tissue in 10 cases. The density could be related to certain histologic features as the amount of connective tissue and extracellular water and the presence of psammoma bodies or lipid storage cells. After administration of contrast a homogeneous, markedly increased, sharply marginated density was seen in most meningiomas. Contrast enhancement appeared to be dependent on the vascularity of the tumour. +/- 50% of the cases demonstrated edema grade II or III. On first interpretation 93% of the meningiomas were recognized as a lesion and a tumour (2 false negative) and 79% as a meningioma (2 false negative and 4 incorrect). Only one false positive C.T. scan was encountered. As a screening method R.N. scanning has about the same accuracy as C.T.; in establishing a definite diagnosis angiography equals C.T.; but C.T. is superior as it combines the diagnostic qualities of both methods.


Doppler haematotachography: problems in interpretation and new applications. The Doppler-echographic examination (HTG) seems to be developing into a screening method for the detection of abnormalities of the large cerebropetal vessels. An appraisal is made of the possibilities and limitations of the HTG based on the literature, a group of 250 patients and several relevant case histories. In general an abnormal HTG curve seems to warrant further diagnostic investigations (angiography). A normal HTG curve not be considered sufficient reason to omit an angiographic examination. A number of new applications of the HTG examination are mentioned.


Siderophages in differentiation of blood in cerebrospinal fluid. This paper evaluates demonstration of siderophages as a method of differentiating blood in CSF. Of 1147 CSF samples, 125 were selected which had been obtained by lumbar puncture from patients with proven intracranial haemorrhage (spontaneous or traumatic) after an exactly known interval since the onset of clinical phenomena (haemorrhage or accident). No siderophages were found within 24 hours; in a few cases they were found on the second or the third day, and after that time they were found in the majority of cases. This is a pathognomonic finding. Collection of CSF in successive portions, qualification of the tap by the clinician as 'traumatic' or 'atraumatic', and demonstration of crenated red cells, were found to be unreliable methods. A modified apparatus for sedimentation is described.


Regulation of gluconeogenesis and ketogenesis during rest and exercise in diabetic subjects and normal men. The splanchnic-hepatic metabolism of glucose, lactate, pyruvate, alanine, glycerol, non-esterified fatty acids (NEFA), ketone bodies and oxygen were investigated in five normal men and six juvenile diabetic subjects at rest and during exercise after an overnight fast. A linear relationship was found between load (arterial concentration multiplied by hepatic blood flow) and splanchnic-hepatic uptake of lactate, pyruvate, glycerol and NEFA. The uptake of alanine was highly sensitive to load, but was also regulated by the concentration of hepatic venous glucagon. The uptake of pyruvate was high in exercising diabetic subjects, who had a high lactate/pyruvate concentration ratio in hepatic venous blood. The rate of uptake of the total measured gluconeogenic precursors was significantly higher in the diabetic group at a given load. The rate of ketogenesis was linearly related to the NEFA load in both groups; however, the rate of ketogenesis was twofold at a given load in the diabetic group. The highest rates of ketogenesis were found coincident with the highest concentrations of glucagon in hepatic venous blood. The observed antiketogenic effect of exercise was due to a decreased load of NEFA, mainly caused by a decrease in the hepatic blood flow.


Role of cyclic AMP in the natriuresis of extracellular fluid volume expansion in the dog. 1. The effect of extracellular volume expansion (ECVE) on renal production of cyclic AMP was evaluated in 19 thyroparathyroidectomized dogs. ECVE was produced by the infusion of Ringer bicarbonate solution at a rate of 2 ml min-1 kg-1 body weight; cyclic AMP was measured in plasma obtained from the aorta and renal vein and in the urine. 2. During the natriuresis of ECVE urinary excretion of cyclic AMP, the clearance of cyclic AMP, net nephrogenous cyclic AMP added both to urine and to the renal vein and hence total nephrogenous cyclic AMP increased significantly. 3. This rise in net production of cyclic AMP and a significant natriuresis by the kidney persisted for 60--90 min after discontinuation of active ECVE and return of renal plasma flow to normal. 4. The results support the notion that an increase in the production of cyclic AMP by the kidney may play a role in the natriuresis of ECVE.