Substrate effects on mitochondrial function and tissue lipids in low-flow hypoxia of isolated perfused rat hearts. A possible causal relationship between tissue FFA contents and the depression in mitochondrial oxidative phosphorylation in myocardial ischaemia has been suggested. To test this hypothesis, the effects of different substrates added to the perfusates of hypoxic, low-flow perfused hearts were examined on oxidative phosphorylation catalysed by mitochondria isolated from such tissue. In an additional series of experiments tissue neutral glyceride and FFA levels were analysed and correlated with changes in mitochondrial function. Mitochondria isolated from hearts with a high tissue FFA content exhibited the lowest ADP/O ratios, RCI and QO2 values. On the other hand, mitochondria isolated from hearts with reduced FFA contents, performed significantly better with respect to these parameters of mitochondrial function studied.


Mass-spectral identification and purification of phycoerythrobilin and phycocyanobilin. The bile-pigment chromophores of C-phycoerythrin (phycoerythrobilin) and C-phycocyanin (phycocyanobilin) were cleaved from their respective proteins with boiling methanol or butan-1-ol. They were purified as dicarboxylic acids by preparative reverse-phase liquid chromatography. Each pigment existed in two principal forms, which were characterized by using 20 pmol samples by proton-transfer chemical-ionization mass spectroscopy. These two principal forms were isomeric species, and all had protonated parent molecular ions with m/e 587, corresponding to a molecular weight of 586.


Characterization of lipid-protein interactions in acetylcholinesterase lipoprotein extracted from bovine erythrocytes. Acetylcholinesterase was released from bovine erythrocytes in hypo-osmotic sodium phosphate buffer. Initially, about 30% of the enzyme was released in a soluble lipoprotein form, and further incubation resulted in the progressive release of the enzyme in a particulate form. Solubilization of the acetylcholinesterase in the particulate fraction with Lubrol WX (2 mg/ml) resulted in the loss of all lipids except a non-exchangeable fraction identified as cardiolipin. Addition of a mixture of erythrocyte phospholipids to the soluble forms and to the Lubrol WX-solubilized enzyme resulted in the formation of particulate forms of the enzyme with increased partial specific volume and Stokes radius, and a break in the Arrhenius plot of the enzyme activity around 20 degrees C. The break in the Arrhenius plot was abolished by treatment of a soluble enzyme preparation with 1.8 M salt (NaCl) in phosphate buffer, conditions that allowed the extraction of cardiolipin from the enzyme by chloroform/methanol. Failure of the high-salt treatment to decrease the Stokes radius made it unlikely that the bound cardiolipin formed a boundary layer or annulus around the protein. It is suggested that cardiolipin is bound to the core of the dimeric protein structure, thereby controlling the acetylcholinesterase activity.


4-pyridylmethyl, a thiol-protecting group suitable for the partial synthesis of proteins. Cysteine is converted into S-4-pyridylmethylcysteine [Gosden, Stevenson & Young (1972) J. Chem. Soc. Chem Commun. 1123-1124] by 4-pyridylmethyl chloride in aqueous propanol at pH 8.3. The derivative is stable to the conditions of total acid hydrolysis. Reduction and alkylation of bovine insulin (pH 8.3, aq. 50% propanol) gives fully S-substituted derivatives in excellent yields. The S-pyridylmethylated A- and B-chains of insulin were separated by gel filtration: each of them has good solubility properties. The pyridylmethyl group is cleaved by electrolysis in a dilute acid medium, pH 2.6, to give reduced chains. They can be recombined to give insulin in the same yield and with the same degree of biological activity as chains which had not been subjected to the protection and de-protection steps. The results indicate that pyridylmethyl satisfactorily meets requirements for partial synthesis and suggest that it warrants more general use.


2-Sulphobenzyl, a new solubilizing and reversible protecting group for cysteine in proteins. Its scope and limitations. S-2-Sulphobenzylcysteine and S-2-(sulphomethyl)benzylcysteine are prepared by alkylation of cysteine with omega-toluenesultone and 2,3-benzo-1,4-butanesultone respectively. Owing to the presence of the sulphonic acid group, these protected cysteine derivatives are extremely water-soluble and are stable to acid hydrolysis. The groups can be removed by treatment with sodium in liquid NH3. Reduction with tributylphosphine and simultaneous alkylation of insulin with toluenesultone under mild conditions (pH 8.3, aq. 50% propanol) gives the fully S-substituted derivatives in excellent yield; they can be separated by isoelectric precipitation of the S-sulphobenzylated B-chain. Treatment of the latter with sodium in liquid NH3 led simultaneously to the removal of the protecting groups and to the well-documented cleavage at the threonine-proline bond which can be prevented by addition of sodium amide. When deprotected A-chain was recombined with B-chain, insulin was isolated in the same yield and with the same degree of biological activity as that in the control experiment.


Kinetic behaviour of acid phosphatase-albumin co-polymers in homogeneous phase and under gel-immobilized conditions. 1. An analysis of the kinetic behaviour of immobilized acid phosphatase (EC 3.1.3.2) layers, gelled on the active surface of an ultrafiltration membrane, was carried out. 2. Two possible forms of such immobilized-enzyme systems were dealt with, namely enzyme-polyalbumin co-gelation through an ultrafiltration process, and enzyme co-polymerization to the same albumin polymers and subsequent gelation. 3. A preliminary analysis was also performed on both the corresponding homogeneous-phase (soluble systems to provide reference kinetics. 4. The main conclusions drawn are: (i) the enzyme-albumin co-polymers show a decrease in specific activity compared with the corresponding free enzyme in both soluble and immobilized forms; (ii) in the homogeneous phase a slight increase in the apparent Michaelis constant was measured for the co-polymerized enzyme compared with the free one, which suggests a decrease in affinity towards substrate; (iii) the activation energy in the immobilized phase is halved, compared with that in the homogeneous phase, which indicates that the combined mass-transfer/reaction step is rate-controlling.


The effect of restricted hydration on the rate of reaction of glucose 6-phosphate dehydrogenase, phosphoglucose isomerase, hexokinase and fumarase. A method is described for the measurement of enzyme activity under xeric conditions. The reaction mixtures had water contents ranging between 0.1 and 0.6g/g of reaction mixture. For glucose 6-phosphate dehydrogenase, hexokinase and fumarase, enzyme activity became detectable (about 0.05% of the fully hydrated rate) when the water content was about 0.2g/g of reaction mixture, and for phosphoglucose isomerase, around 0.15g/g of reaction mixture. With the water content raised to 0.3g/g of reaction mixture the reaction rates were only increased to 0.1-3% of the fully hydrated rate. When the combined rates for phosphoglucose isomerase and glucose 6-phosphate dehydrogenase were measured, reasonable agreement was found between the experimental data and those calculated from the individual experimentally determined rates on the assumption that diffusion was not further limiting. A method was devised for measuring the diffusion coefficients of low-molecular-weight substances in solutions having low water contents. The diffusion coefficients of riboflavin in sorbitol solution decreased by about 100-fold when the water content of the latter was reduced from 3 to 0.25g/g of sorbitol. It is concluded that to detect enzyme activity a certain minimal amount of water is required and that above this minimum the rate is still restricted by diffusion limitation. The relevance of the results to the physical state of water in reaction mixtures and to metabolism in seeds and spores in xeric conditions is discussed.


Binding of ethidium bromide and quinacrine hydrochloride to nucleic acids and reconstituted nucleohistones. Studies of binding of ethidium bromide and quinacrine hydrochloride to native DNA at low ionic strength indicate that for both compounds the binding is selective, with about one binding site for about four nucleotides. Annealing of unfractionated histones to DNA by a salt-gradient dialysis method slightly decreases the binding of the dyes to DNA. Similar observations made with reconstituted preparations by using individual histone fractions reveal that the arginine-rich histones (histones H3 and H4) are most effective in decreasing the binding. The binding studies with ethidium bromide at high ionic strength and with denatured DNA show that strong dye binding to DNA is strongly dependent on the ionic strength and on the secondary structure of DNA. The histones are not effective in decreasing the dye binding under conditions of high ionic strength. The results are consistent with the observations [Oliver & Chalkley (1974) Biochemistry13, 5093-5098; Axel, Melchoir, Sollner-Web & Felsenfield (1974) Proc. Natl. Acad. Sci. U.S.A.71, 4101-4105] that histones form some kind of surface structures on DNA through non-specific interactions and [Kornberg & Thomas (1974) Science184, 865-868; Kornberg (1974) Science184, 868-871; D'Anna & Isenberg (1974) Biochemistry13, 4992-4997; Vandegrift, Serra, Marve & Wagner (1974) Biochemistry13, 5087-5092] that the tendency of arginine-rich histones to aggregate may be an important factor in determining the structure of chromatin.


Neutral glycosphingolipids and gangliosides of human lung and lung tumours. In order to help determine whether alterations of the profiles of glycosphingolipids occur consistently in human tumours, the neutral glycosphingolipids and gangliosides of nine lung tumours (one adenocarcinoma, four squamous cell, two mixed adeno-squamous cell, one large cell and one oat-cell carcinomata) were analysed. The control tissue consisted of adjacent lung; it contained neutral glycosphingolipids corresponding in properties to glucosyl-, lactosyl-, globotriaosyl- and globotetraosyl-ceramides. All of the tumours also contained these four neutral glycosphingolipids. However, in addition, five of the tumours (two of the squamous, the large cell and the two mixed adeno-squamous cell carcinomata) contained neutral glycosphingolipids corresponding in properties to lactotriaosyl- and neolactotetraosyl-ceramides; these same tumours also exhibited higher amounts of lactosylceramide than the other tumours analysed. Both of the two former neutral glycosphingolipids and very substantial amounts of the latter neutral glycosphingolipid were detected in pneumonic lung and in polymorphonuclear leucocytes; it thus appears possible that these particular compounds were derived from these latter cells rather than from the tumour cells. The ganglioside patterns of the tumours were almost equivalent in complexity to that exhibited by the control lung tissue. This study shows that the profiles of two major classes of glycosphingolipids (neutral glycosphingolipids and gangliosides) occurring in lung tumours are almost as complex as those of the parent tissue, a finding in contrast with the notably simplified patterns of these lipids found in many cancer cells grown in vitro. It also suggests that when lactotriaosyl- and neolactotetraosyl-ceramides and high amounts of lactosylceramide are detected in human tumours, the possibility must be considered that these compounds are derived from polymorphonuclear leucocytes.


Interferon effect on ribosomal ribonucleic acid related to chromosome 21 ploidy. Antiviral and cell-growth-inhibitory activities of human interferon were shown to be related to the activity of a gene or genes present on chromosome 21. The 18s rRNA is vital to cell growth; it is capable of a viral-mRNA-recognition function and it is coded for by genes a portion of which are present on chromosome-21. A previously reported ability of human interferon to affect rRNA metabolism is characterized by a decrease in the sucrose-gradient-peak ratio of radiolabelled 28S to 18S rRNA in extracts from the cytoplasm of interferon-treated human fibroblasts. In the present report, interferon dose-response curves are presented demonstrating a direct relationship between a decrease in this ratio and interferon concentrations in the media. By using this virus-independent cytoplasmic rRNA assay, eight human fibroblast lines, differing in chromosome 21 ploidy, were tested for sensitivity to human interferon. Two monosomy-21, two euploid-21 and four trisomy-21 cell lines were tested. The monosomy-21 cell populations were significantly less sensitive to interferon than the other six cell types tested. Of the cell lines tested, the most sensitive, by a wide margin, was a trisomy-21 line. Trisomy-21 cell monolayer sensitivity, however, varied widely within the range from normal to supersensitive. These observations suggest that interferon's ability to affect rRNA metabolism is related to the activity of a gene or genes present on chromosome 21.


Changes in intermediate haemoglobins during methaemoglobin reduction by NADPH-flavin reductase. The changes in intermediate haemoglobins produced during methaemoglobin reduction by NADPH-flavin reductase were analysed by an isoelectric-focusing method. The alpha 3+ beta 2+ and alpha 2+ beta 3+ valency hybrids were observed as intermediate haemoglobins and changed consecutively with time during the reaction. On the basis of the analyses, the course of methaemoglobin reduction was found to involve two different pathways: (1) methaemoglobin kappa+1 leads to alpha 3+ beta 2+ kappa+2 leads to oxyhaemoglobin; (2) methaemoglobin kappa+3 leads to alpha 2+ beta 3+ kappa+4 leads to oxyhaemoglobin. The reaction rate constants of each phase (kappa+1--kappa+4) were also estimated. The addition of inositol hexaphosphate to the reaction mixture did not affect the overall reaction. The mechanism of methaemoglobin reduction by NADPH-flavin reductase is discussed on the basis of these results.


Effects of modification of the tyrosine residues of bacteriorhodopsin with tetranitromethane. Treatment of the purple membrane of Halobacterium halobium with tetranitromethane led to modification of tyrosine residues. Modification of more than 3-4 tyrosine residues per bacteriorhodopsin monomer caused a decrease in the light-induced proton-pumping ability of purple membrane in synthetic lipid vesicles, loss of the sharp X-ray-diffraction patterns characteristic of the crystal lattice, loss of the absorbance maximum at 560 nm, and change in the buoyant density of the membrane. No modification of lipid was detected. These changes were interpreted as a gradual denaturation of the protein component such that when 8-9 tyrosine residues are modified, no proton pumping is observed. Modification of less than 3-4 tyrosine residues with tetranitromethane caused an increse in light-induced proton pumping. It was possible to generate partly modified purple membrane which had completely lost the property of diffracting X-rays into the sharp pattern observed with native purple membrane, but which still retained the ability to pump protons in a vectorial manner. Retention of crystal lattice is not essential for proton pumping.


The isolation, characterization and partial sequence of a peptide rich in glutamic acid and aspartic acid (HGA-2 peptide) from calf thymus non-histone chromosomal protein HMG 2. Comparison with a similar peptide (HGA-1 peptide) from calf thymus non-histone chromosomal protein HMG 1. A 41-residue peptide (HGA-2) containing a continuous sequence of 35 glutamic and aspartic residues was isolated from non-histone chromosomal protein HMG 2. This highly acidic peptide is compared with a similar peptide (HGA-1) isolated from non-histone chromosomal protein HMG 1.


Biosynthesis of a 7-alpha-methoxycephalosporin. Incorporation of molecular oxygen. A 7-alpha-methoxycephalosporin containing a carbamoyloxymethyl substituent at C-3 (cephamycin C) has been isolated from the extracellular fluid of an aqueous suspension of Streptomyces clavuligerus shaken in the presence of 18O2. The cephalosporin has been converted into its N-acetyl dimethyl ester and the distribution of 18O in the latter determined by chemical-ionization mass spectrometry. The results indicate that the oxygen atom of the methoxy group, as well as that linked to the exocyclic methylene group at C-3, is derived from molecular O2.


Sequential degradation of a chondroitin sulphate trisaccharide by lysosomal enzymes from embryonic-chick epiphysial cartilage. The disulphated trisaccharide D-N-acetylgalactosamine sulphate-beta-D-glucuronic acid-beta-D-N-acetylgalactosamine sulphate prepared from 35S- or 14C-labelled chondroitin sulphate was incubated with a preparation of lysosomal enzymes from embryonic-chick epiphysial cartilage. Degradation was demonstrated by analysis of the reaction products. By use of the appropriate intermediate products as substrates, in conjunction with specific enzyme inhibitors, it was shown that the degradation proceeded sequentially from the non-reducing end. It was initiated by sulphatase (preferentially hydrolysing sulphate ester groups at the 6-position), followed by beta-N-acetylgalactosaminidase and beta-glucuronidase, converting the substrate into monosaccharides and inorganic sulphate. The latter enzyme preferentially attacked disaccharides carrying their sulphate ester group at C-4 of the hexosamine residue. Generation of chondroitin sulphate oligosaccharides may occur by the action of an endoglycosidase, previously demonstrated in embryonic-chick cartilage. Endo- and exo-enzymes may thus form a functional unit in lysosomal degradation of chondroitin sulphate.


C1 inhibitor-dependent dissociation of human complement component C1 bound to immune complexes. The interaction of C1 inhibitor with complement component C1 bound to immune complexes was examined by using 125I-labelled C1 subcomponents. The inhibitor binds rapidly to subcomponent C1s, and more slowly to subcomponent C1r. Formation of the C1r-C1 inhibitor complex causes rapid dissociation of subcomponents C1r and C1s from the antibody-antigen-component C1 aggregate. The rate and extent of this release are proportional to C1 Inhibitor concentration and are also dependent on ionic strength. Results obtained with purified C1 Inhibitor, plasma or serum as source of C1 Inhibitor are all closely comparable. Only slight dissociation of subcomponent C1q is observed under the same range of conditions. The implications of the release phenomenon are discussed in relation to the structure of component C1 and the possibility of differential turnover of C1 subcomponents.


The formation and distribution of bilirubin monoglucuronide and diglucuronide in rat liver slices. 1. Bilirubin conjugation in rat liver slices was reassessed by using analysis of ethyl anthranilate azopigments to estimate separately the formation of bilirubin mono- and di-glucuronides. 2. Conjugation in slices resembles the situation in vivo more closely than does microsomal conjugation, in that diglucuronide is formed in appreciable quantity. 3. Both bilirubin mono- and di-glucuronides were present in slices in approximately equal amounts, but the monoglucuronide was the major product found in the incubation medium. 4. These results are discussed in relation to recent theories on the relationship between bilirubin mono- and di-glucuronide formation in vivo.


The atypical velocity response by  pyruvate carboxylase to increasing concentrations of acetyl-coenzyme A. An investigation was made of the interaction of pyruvate carboxylase with its allosteric effector, acetyl-CoA, and the velocity profile of the deacylation of acetyl-CoA as a function of acetyl-CoA concentration indicated that this ligand does not bind to this enzyme in a positive homotropic co-operative manner. An examination was therefore made of the factors that contribute to the sigmoidicity of the rate curves obtained for pyruvate carboxylation with various concentrations of acetyl-CoA. Hill coefficients for acetyl-CoA obtained with both sheep and chicken liver pyruvate carboxylases were found to be dependent on the fixed pyruvate concentration used in the assay solution. Thus, by varying the acetyl-CoA concentration, the degree of saturation of the enzyme by pyruvate was also changed. A further consequence of non-saturating concentrations of pyruvate was that the non-productive hydrolysis of the enzyme- carboxybiotin complex increased, resulting in an under-estimate of the reaction velocity measured by oxaloacetate formation. Another factor contributing to the sigmoidicity is that, at non-saturating concentrations of acetyl-CoA, the enzyme undergoes inactivation upon dilution to low protein concentrations, again resulting in an under-estimate of the reaction velocity. Under conditions where none of the above factors was operating and the only effect of varying acetyl-CoA concentrations was to alter the proportion of the enzyme catalysing the carboxylation reaction at acetyl-CoA-dependent and -independent rates, the sigmoidicity of the acetyl-CoA velocity profile was completely eliminated.


Analysis of the ratio of alpha- to beta-globin and globin messenger ribonucleic acid content of fractionated rabbit erythroid bone-marrow cells. The ratio of alpha- to beta-globin mRNA was measured by hybridization of a constant amount of highly purified alpha- or beta-globin cDNA (complementary DNA) with increasing amounts of RNA in the range up to 20% cDNA hybridization, where an essentially linear reaction is obtained. Statistical analysis indicates that the ratio of alpha- to beta-globin can be measured within a maximal error of +/- 0.3 and in most cases is better than +/- 0.15. Under these conditions there is no significant deviation from the ratio of 1.3 in the alpha- to beta-globin mRNA ratio of RNA isolated from erythroid cells rich in pronormoblasts through to reticulocytes. If the ratio of alpha- to beta-globin mRNA exceeded 1.7 or was less than 0.9 in pronormoblasts, it would be detected in these experiments. The overall globin mRNA content increases to a maximal value in the fractions rich in basophilic normoblasts of 30,000--50,000 molecules/cell. However, the accuracy of these determinations is not as great as for the ratio determinations, and no significant deviations were seen except in the cells rich in pronormoblasts, which contained less globin mRNA than the later stages.


Proteoglycans of the intervertebral disc. Homology of structure with laryngeal proteoglycans. The structure of the proteoglycans from normal pig nucleus pulposus and relatively normal human annulus fibrosus and nucleus pulposus was investigated in detail and the results were compared with the current structural model of proteoglycans of hyaline cartilage. Like proteoglycans of cartilage, those of intervertebral disc contain keratan sulphate and chondroitin sulphate attached to a protein core; they are able to aggregate to hyaluronic acid; the protein core likewise has three regions, one lacking glycosaminoglycans, another rich in keratan sulphate and a third region rich in chondroitin sulphate. However, disc proteoglycans contain more keratan sulphate and protein and less chondroitin sulphate and are also considerably smaller than cartilage proteoglycans. In proteoglycans of human discs, these differences appeared to be due principally to a shorter region of the core protein bearing the chondroitin sulphate chains, whereas in proteoglycans of pig discs their smaller size and relatively low uronic acid content were due to shorter chondroitin sulphate chains. There were subtle differences between proteoglycans from the nucleus and annulus of human discs. In the latter a higher proportion of proteoglycans was capable of binding to hyaluronate.


Proteoglycans of the intervertebral disc. Absence of degradation during the isolation of proteoglycans from the intervertebral disc. Proteoglycans extracted with 4M-guanidinium chloride from pig intervetebral discs, and purified by equilibrium density-gradient centrifugation in CsCl, were of smaller hydrodynamic size than those extracted and purified in the same way from the laryngeal cartilage of the same animal. Whether this difference in size arose from degradation during the extraction and purification of the proteoglycans of the disc was investigated. Purified proteoglycans labelled either in the chondroitin sulphate chains or in the core protein were obtained from laryngeal cartilage by short-term organ culture. These labelled proteoglycans were added at the beginning of the extraction of the disc proteoglycans, and labelled cartilage and unlabelled disc proteoglycans were isolated and purified together. There was no appreciable loss of radioactivity after density-gradient centrifugation nor decrease in hydrodynamic size of the labelled cartilage proteoglycans on chromatography on Sepharose 2B, when these were present during the extraction of disc proteoglycans. It is concluded that disc proteoglycans are intrinsically of smaller size than cartilage proteoglycans and this difference in size does not arise from degradation during the extraction.


The kinetics of rat liver and heart mitochondrial beta-hydroxybutyrate dehydrogenase. The kinetic mechanisms of the beta-hydroxybutyrate dehydrogenase from rat heart and liver mitochondria were investigated. Both enzymes, show an Ordered Bi Bi mechanism and there are no major differences in the kinetic constants. In both cases, the solubilized enzyme, re-activated with phosphatidylcholine, shows kinetic properties very similar to those of the enzyme bound to the mitochondrial membrane.


The multiple forms of alpha-D-mannosidase in human plasma. The acidic alpha-D-mannosidase in human plasma closely resembles liver acidic alpha-D-mannosidase in its affinity for concanavalin A-Sepharose, molecular weight and resolution into multiple components on DEAE-cellulose. A combination of chromatography on concanavalin A-Sepharose and gel filtration on Sephadex G-200 and Sepharose 6B suggests that four forms of intermediate alpha-D-mannosidase, which differ either in their molecular weight of affinity for concanavalin A, exist in human plasma. A practical classification and nomenclature for the multiple forms of intermediate alpha-D-mannosidase in plasma based on molecular weight and affinity for concanavalin A is proposed. Multiple forms of intermediate alpha-D-mannosidase were also observed by ion-exchange chromatography on DEAE-cellulose, but there was not a simple correlation between these forms and those obtained with the other separation procedures. The form of intermediate alpha-D-mannosidase least abundant in plasma, approx. 7% of the activity, has very similar properties to the neutral alpha-D-mannosidase in human liver. In contrast, the other three forms of intermediate alpha-D-mannosidase, which account for over 90% of the activity, do not appear to be present in liver, except perhaps in trace amounts.


Characterization of domains obtained from a mollusc haemocyanin by limited proteolytic digestion. The haemocyanin from the freshwater gastropod Lymnaea stagnalis was digested with proteolytic enzymes under conditions where it existed as whole (native) molecules (mol.wt. approx. 9 X 10(6)), or as one-tenth molecules. Digestion of whole molecules yielded a fragment of mol.wt. approx. 110,000 believed to correspond to the 'collar' of the molecule, and an aggregate some 20--30 times the size of the original native molecule formed by end-to-end polymerization of the molecule after removal of the collar. Digestion of one-tenth molecules yielded a mixture of products that could be separated into three fractions by gel filtration. Analysis of these by sodium dodecylsulphate/polyacrylamide-gel electrophoresis revealed that they typically contained two or three components. The collar fragment was present as a component of the intermediate-molecular-weight fraction, and it dissociated on sodium dodecyl sulphate/polyacrylamide gels to give two bands corresponding to apparent mol.wts. 65,000 and 60,000. The c.d. spectra of the separated fractions were recorded and fitted with Gaussian curves by a computer procedure. The fractions each possessed distinct c.d. spectra, by which they could be identified: the collar-fragment c.d. and absorption spectra showed the most striking differences compared with those of the other fragments. The results were interpreted in terms of the postulated existence, within the haemocyanin molecule, of multi-domain structures, each comprising a single polypeptide chain of mol.wt. 200,000--300,000.


Purification and characterization of a phospholipase A2 from the venom of the coral snake, Micrurus fulvius microgalbineus (Brown and Smith). A phospholipase A2 was purified from the Mexican coral snake Micrurus fulvius microgalbieus (Brown and Smith). Gel filtration of the soluble crude venom on Sephadex g-50 resolved five fractions, of which fraction II had 98% of the total phospholipase activity. This fraction was rechromatographed on a CM-cellulose column that resolved eight fractions, four of which had an important phospholipase activity. The first fraction (II-1) was homogeneous by polyacrylamide-gel electrophoresis and displayed a phospholipase specific activity of 920 units/mg of protein. The apparent molecular weight as determined by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis was approx. 14000. The amino acid analysis revealed the presence of 119 amino acid residues, with 12 half-cystines. the N-terminal sequence was shown to be Ser-Leu-Leu-Asx-Phe-Lys-Asx-Met-Ile-Glu-Ser-Thr..., which is homologous with that of phospholipases from other snake venoms.


Amino acid sequence of the N-terminal non-collagenous segment of dermatosparactic sheep procollagen type I. The non-collagenous N-terminal segment of type I procollagen from dermatosparactic sheep skin was isolated in the form of the peptide Col 1 from a collagenase digest of the protein. The peptide has a blocked N-terminus, which was identified as pyrrolid-2-one-5-carboxylic acid. Appropriate overlapping fragments were prepared from reduced and alkylated peptide Col 1 by cleavage with trypsin at lysine, arginine and S-aminoethyl-cysteine residues and by cleavage with staphylococcal proteinase at glutamate residues. Amino acid sequence analysis of these fragments by Edman degradation and mass spectrometry established the whole sequence of peptide Col 1 except for a peptide junction (7--8) and a single Asx residue (44), and demonstrated that peptide Col 1 consists of 98 amino acid residues. The N-terminal portion of peptide Col 1 (86 residues) shows an irregular distribution of glycine, whereas the C-terminal portion (12 residues) possesses the triplet structure Gly-Xy and is apparently derived from the precursor-specific collagenous domain of procollagen. The central region of the peptide contains ten cysteine residues located between positions 18 and 73 and shows alternating polar and hydrophobic sequence elements. The regions adjacent to the cysteine-rich portion have a hydrophilic nature and are abundant in glutamic acid. The data are consistent with previous physicochemical and immunological evidence that distinct regions at the N- and C-termini of the non-collagenous domain possess a less rigid conformation than does the central portion of the molecule.


Evidence for an oleoyl phosphatidylcholine desaturase in microsomal preparations from cotyledons of safflower (Carthamus tinctorius) seed. 1. [14C]Oleoyl-CoA was metabolized rapidly and essentially completely by microsomal preparations from developing safflower (Carthamus tinctorius) cotyledons, and most of the [14C]oleate was incorporated into 3-sn-phosphatidylcholine. 2. In aerobic reaction mixtures containing NADH2 the [14C]oleate in 3-sn-phosphatidylcholine was converted into [14C]linoleate without any change in the specific radioactivity of the lipid. Over a 60 min incubation period the extent of conversion of [14C]oleoyl phosphatidylcholine into [14C]linoleoyl phosphatidylcholine was generally greater than 60%. The rate of desaturation of endogenous [14C]oleoyl phosphatidylcholine labelled from [14C]oleoyl-CoA was much greater that of exogenous [14C]dioleoyl phosphatidylcholine the specific radioactivity of the oleoyl moiety of the lipid remained constant, indicating that labelled and unlabelled oleate were desaturated at the same rate. On this assumption an initial rate of desaturation of about 15 nmol of oleate desaturated/min per mumol of 3-sn-phosphatidylcholine was estimated. 4. [14C]Oleate esterified at positions 1 and 2 of both endogenous and exogenous 3-sn-phosphatidylcholine was desaturated. 5. Attempts to demonstrate the presence of an oleoyl-CoA desaturase in safflower microsomal fractions by the appearance of linoleoyl-CoA in reaction mixtures were inconclusive.


The reaction of menadione with haemoglobin. Mechanism and effect of superoxide dismutase. 1. Menadione was found to react with both the haem groups and the beta-93 thiol groups of haemoglobin. 2. It oxidized the haem groups of oxyhaemoglobin, giving mainly methaemoglobin and a smaller amount of haemichrome. The reaction rate was decrease in the presence of catalase and markedly accelerated in the presence of superoxide dismutase. It is proposed that the overall reaction involves the initial reversible formation of methaemoglobin and the semiquinone, and that the effect of superoxide dismutase is to prevent the reverse reaction, by removing superoxide and hene O2-. E.s.r. evidence for the information of the semi-quinone and its reactions is presented. 3. The reaction of menadione with the beta-93 thiol groups of haemoglobin appeared to be similar to that with other thiols, forming the 3-thioether derivative of menadione, but it was also accompanied by reduction of methaemoglobin. This reduction was prevented by superoxide dismutase, but appeared to be caused by the semiquinone radical, which was produced as an intermediate. 4. Reduced glutathione functioned only to a limited extent as a scavenger of the menadione semiquinone. Its main reaction was directly with menadione to form the thioether. Ascorbate was a more efficient scavenger, and accelerated the oxidation of oxyhaemoglobin by menadione. 5. The significance of these findings in relation to menadione-induced erythrocyte haemolysis is discussed.


The determination of binding parameters when the total and free substrate concentrations are not approximately equal. By using a standard graphical method values of Km and V may be found that are independent of the conditions and assumptions that the total substrate concentration approximates to its free concentration and that Km is much larger than the enzyme concentration. The procedure is also applicable to the determination of equilibrium binding parameters of a ligand to a macromolecule.


The separation of sheep liver cytoplasmic and mitochondrial aldehyde dehydrogenases by isoelectric focusing, and observations on the purity of preparations of the cytoplasmic enzyme, and their sensitivity towards inhibition by disulfiram. Preparations of sheep liver cytoplasmic aldehyde dehydrogenase obtained by published methods were found by analytical isoelectric focusing in the pH range 5--8 to contain 5--10% by weight of the mitochondrial aldehyde dehydrogenase. Under the conditions used the pI of the cytoplasmic enzyme is 6.2 and that of the mitochondrial enzyme 6.6. The mitochondrial enzyme can be removed from the preparation by selective precipitation of the cytoplasmic enzyme with (NH4)2SO4. Kinetic experiments and inhibition experiments with disulfiram show that the properties of the two sheep liver enzymes are so different that the presence of 10% mitochondrial enzyme in preparations of the cytoplasmic enzyme can introduce serious errors into results. Our results suggest that the presence of 10 microM-disulfiram in assays may completely inactivate the pure cytoplasmic enzyme. This result is in contrast with a previous report [kitson (1978) Biochem. U. 175, 83--90].


Co-ordinate regulation of ethanolamine kinase and phosphoethanolamine cytidylyltransferase in the biosynthesis of phosphatidylethanolamine in rat liver. Essential-fatty acid deficiency produces a 52% increase in the rate of phosphatidyl-ethanolamine synthesis in rat liver as calculated from results obtained in vivo [Trewhella & Collins (1973) Biochem. Biophys. Acta 296, 34--50]. This flux change was used to test the possible regulatory roles of ethanolamine kinase and of phosphoethanolamine cytidylyltransferase, which are rate-limiting enzymes of the cytidine pathway for the synthesis of phosphatidylethanolamine [Infante (1977) Biochem. J. 167, 847--849]. The results show that essential-fatty acid deficiency produces 50% and 53% increases respectively in the specific activity of these enzymes, accounting for the increased rate of phosphatidylethanolamine synthesis produced by this dietary insufficiency. This evidence leads to the conclusion that ethanolamine kinase and phosphoethanolamine cytidylyl-transferase have co-ordinated regulatory roles in the flux control of the cytidine pathway, and its sphinganine 1-phosphate lyase branch reaction, for the synthesis of phosphatidylethanolamine.


Early complement component depletion and mixed cryoglobulinemia in a "healthy" family. A family containing two apparently healthy brothers with profound early complement component depletion (C1, C4, C2) and mixed IgG-IgM polyclonal cryoglobulins was studied. The cryoglobulins possessed rheumatoid factor activity and depleted early complement components in normal human serum. Circulating immune complexes could not be detected by utilizing standard methods. The phenomenon was not HLA-linked. This study demonstrates the familial occurrence of a chronic hypocomplementemic state associated with cryoglobulinemia in clinically normal subjects.


Does corticosteroid therapy affect the survival of patients with systemic lupus erythematosus? The hypothesis that corticosteroid therapy is responsible for the striking improvement in survival of patients with systemic lupus erythematosus (SLE) was investigated with two approaches: Of 250 published papers on SLE, 52 were chosen for the first analysis because they included sufficient information on diagnostic criteria and survival but were not limited to patients selected for a particular target organ. From each article percent survival by series, average duration to death, and 1 and 5 year survival curves were abstracted. Each statistic showed linear improvement in survival since the 1930s without a significant change (P greater than 0.10) in slope for the time period following the introduction of corticosteroids. The second analysis examined the effect of corticosteroid therapy on 142 patients with SLE followed at the Massachusetts General Hospital between 1922 and 1966. Although the steroid use was conservative, the patients' survival, from year of entry, was comparable to the 52 abstracted series. A prognostic index was used to stratify patients admitted in the steroid era (after 1950) for life table analysis of survival with and without steroids. Steroids had no discernible effect on overall survival in low and medium risk groups. Use of steroids was associated with improved survival among high rish patients (P less than 0.05).


IgG rheumatoid factor. Relationship to seropositive rheumatoid arthritis and absence in seronegative disorders. IgG rheumatoid factor was detected in the sera of the majority of patients with seropositive rheumatoid arthritis. Values suggestive of IgG rheumatoid factor were noted in one-quarter of patients with seronegative inflammatory arthropathies. These determinations were always low and correlated with elevated IgG concentrations, suggesting nonspecific adherence of IgG rather than a true antigen-antibody reaction. In support of this conclusion, nonrheumatoid factor IgG was capable of concentration-dependent nonspecific adsorption to the solid phase. IgG, but not IgM, rheumatoid factor corresponded with disease activity in patients with seropositive rheumatoid arthritis, suggesting that IgG rheumatoid factor may be important in the pathogenesis.


The rarity of Felty's syndrome in blacks. Evidence is presented that Felty's syndrome (FS) is rare among black patients with rheumatoid arthritis (RA). All of 12 patients with FS seen at Parkland Memorial Hospital, Dallas, Texas betwen 1964 and 1978 were white. During this period 52% of patients admitted to the Parkland medical service were black and 31% of patients dischargd with a diagnosis of RA were black. The number of expected black cases of FS on the basis of the racial distribution of hospitalized patients with RA was 3.7 (P is less than 0.02 when the zero incidence in blacks was compared with the expected incidence). All 7 cases of FS observed at Charity Hospital, New Orleans, Louisiana between 1968 and 1978 were also white. During this period, 65% of patients discharged with a diagnosis of RA were black, and the number of expected black cases of FS was 4.5 (P is less than 0.001). These findings suggest a genetic basis for the development of leukopenia and splenomegaly in RA patients.


Neurogenic regulation of lipid metabolism in the rabbit. A mechanism for the cholesterol-lowering effect of acupuncture. Plasma and aortic cholesterol levels are known to fall significantly after needling at the specific point which is innervated by a branch of the deep peroneal nerve. Different types of stimulation were applied to the specific nerve, using solely needling, injection of normal saline with needling and injection of a small dose of novocain with needling at the specific point in several groups of rabbits fed a high cholesterol diet for two or three weeks. Significant decreases in plasma and aortic cholesterol were obtained in the combined acupuncture group after one or two weeks of acupuncture. When the specific point was blocked by injection of a larger dose of novocain or the deep peroneal nerve was cut, then needling at the specific point lost its hypocholesterolemic effect. It is thus suspected that stimulation of sensory receptors of the deep peroneal nerve at the specific point may be the mechanism of the cholesterol-lowering action of acupuncture.


Some particular aspects of the microarchitecture of human coronary arteries. The following light-microscopic features characterize the microarchitecture of human coronary arteries compared with other organ arteries of muscular type and similar size: (a) the rapid development of a diffuse thickened intima which in adolescents becomes the main layer of the coronary artery wall; (b) the slow post-natal increase in the number of circularly oriented rows of smooth muscle cells of the media; (c) the occurrence of numerous longitudinal muscle columns, particularly in arterial segments and branches which run from the base to the apex of the heart; (d) the complex organization of branching points, including the presence of large pads or cushions, a very thick intima and many longitudinal muscle columns, some of which exhibit a spiral course at the level of the branch mouths with the formation of muscle rings round the entrance orifices.


Study of fibrous plaques occurring in the coronary arteries of children. The pathology of the coronary arteries of children, in relation to atherosclerotic involvement, appeared as a pathology of the main emergence areas and branching points of the left coronary artery and particularly of the anterior descending artery. The first atherosclerotic lesions occurred as non-raised fibrous plaques in 2% of children 6--10 years old and in 4% of children and juveniles 11--15 years old. In the latter age group fatty streaks and gelatinous plaques were also seen in 6% of the subjects; in their appearance they lag behind fibrous plaques by 5--8 years. Indirect evidence was obtained that some branch pads or cushions might be converted into fibrous plaques. The transitional aspects included edema, histolysis (elastolysis, collagenolysis, ground-substance depletion and degenerative cell changes), followed by reorganization and homogeneization of the pre-existing heterogeneous microarchitecture; in a final stage the prevalent processes seemed to be the nodular proliferation of smooth muscle cells and the abundant neoformation of collagen fibers. In essence the results show that in atherosclerosis the coronary arteries are involved in a different sequence and histogenetic pattern than the aorta.


The fatty acid pattern of triglycerides and FFA in serum of spontaneously hypertensive rats (SHR). The fatty acid patterns of serum triglycerides and FFA in SHR and in normotensive controls aged 4, 8 and 26 weeks were estimated by gas-liquid chromatography. In serum triglycerides of SHR, the percentage of linoleic acid (C18:2) was lower and the content of arachidonic acid (C20:4) higher than in age-matched control animals. A continuous increase in palmitic (C16) and linoleic acids as well as a decrease in arachidonic acid has been found with advancing age, the most striking differences existing between 4- and 8-week-old animals, i.e. before onset of arterial hypertension in SHR. In the pre-hypertensive stage, the percentage of arachidonic acid was about 3 times as high as in later stages in SHR. This gradation was, however, even more pronounced in normotensive control rats. The C18:2/C20:4-ratio of triglycerides was lower in SHR but increased with age in both groups reaching a 5--8-fold level. Similar behavior of the FFA fatty acid pattern was less marked. Alterations in levels of linoleic and arachidonic acids are of interest because of their pathogenic role as precursors of prostaglandins in the development of genetically spontaneous hypertension in rats. The results are discussed in connection with the hypotensive effect of a linoleic acid-rich diet recently reported in hypertensive rats.


Early increase of a new platelet coagulant activity in rats fed a thrombogenic diet. A recently described platelet coagulant activity (factor X activating activity), whose pathophysiological significance is as yet unknown, was studied in rats fed a fat-rich (thrombogenic) diet for 1, 2 and 7 weeks and compared to rats fed normal laboratory chow. Whatever the duration of the special feeding period, a highly significant shortening of the clotting time, used for measuring this activity, was observed. When the platelet coagulant activity of individual "fat-fed"rats was quantitated by reference to that of individual control animals, we found a mean increase of 350% (n = 9) after one week and 267% (n = 3) after two weeks of dietary treatment. Partial thromboplastin time, thrombin time and soluble fibrin monomer complexes did not differ in control and treated animals. It seems that platelet coagulant activity, as measured in our test system, is one of the first laboratory parameters to be modified by fat-rich diets. These findings may be relevant to an understanding of the role of platelet coagulant activities other than platelet factor 3 in thrombotic phenomena.


Increased growth of human fibroblasts and arterial smooth muscle cells from diabetic patients related to diabetic serum factors and cell origin. Fibroblasts from 3 diabetic patients (DF) grew faster, resulting in higher cell counts in the stationary phase than fibroblasts from 3 age-matched healthy volunteers (NF). This difference was apparent when DF or NF were cultured in either diabetic (DS) or normal serum (NS). Diabetic serum increased growth of both DF and NF compared with normal serum. Total protein content per plate paralleled the increase of cell number per plate in relation to cell origin and serum type. DS increased growth and total protein per plate in the arterial smooth muscle cell line from a non-diabetic patient in a way similar to in DF and NF. It is concluded that increased growth of DF in vivo could result in an increased turnover of vascular cells with a shortened replicative lifespan, leading to an accumulation of basal lamina. This effect would be even further accentuated by exposure of DF to DS. Taken together with the increased protein synthesis the accelerated development of diabetic angiopathy could be the final consequence.


Lipoprotein lipase activity in human adipose tissue during induced hypertriglyceridaemia. Needle biopsies of adipose tissue and blood samples were obtained before and at short intervals after a "bolus" injection of 10% Intralipid. Lipoprotein lipase activities were measured in acetone--ether extracts of the tissue samples. Levels of serum triglyceride began to fall less than 5 min after the injection of the Intralipid with a half-life of 20 min. During this time interval, no significant changes were observed in the activities of lipoprotein lipase in adipose tissue. A patient with a severe hypertriglyceridaemia (Type V) underwent plasma exchange with a reduction in serum triglyceride levels from 11 to 4 mm/l. There was a parallel fall in adipose tissue lipoprotein lipase activity. We conclude that lipoprotein lipase in adipose tissue is unaltered during experimental hypertriglyceridaemia and that the activity of the enzyme in adipose tissue is probably not reduced as a secondary feature of an elevated plasma triglyceride level.


Adipose cell size in spontaneously hypertensive rats (SHR). The size of adipose cells in spontaneously hypertensive rats (SHR) and normotensive controls has been evaluated at 4, 8 and 26 weeks of age. Age-matched groups showed significant differences only in 8-week-old rats, but this can be explained by the lower body weight of SHR. In both groups of animals fat cell size varies with body weight (r = 0.965 in SHR and r = 0.863 in normotensive rats) independent of the stage of hypertension. The regression lines are not significantly different. Thus, no evidence of enlarged adipocytes in SHR has been obtained.


Properties of flavins where the 8-methyl group is replaced by mercapto- residues. Sulfur functions in position 8 of the flavin nucleus give rise to new modified flavin derivatives, which should prove useful as probes of the flavin binding domains of flavoproteins. Here, we report on some properties of 8-nor-8-alkylmercaptoflavins and 8-nor-8-mercaptoflavin which are readily formed by nucleophilic displacement by alkylmercaptides or sulfide, with 8-nor-8-chloroflavins as starting material. The new flavins are characterized by extensive shifts in spectral properties, with very high extinction coefficients. 8-nor-8-mercaptoriboflavin is easily and reversibly converted to its (-S-S-) dimer. Oxidation of the sulfur group by peracids forms first sulfoxides and then sulfones, in which the characteristic usual flavin spectrum is regained. A comparison of 8-SR-8-nor-flavins with 8-OR-8-nor-flavins (Ghisla, S., and Mayhew, S.G. (1976) Eur. J. Biochem 63, 373-390) indicates that in both classes of compounds, optical properties, ionization constants, and oxidation-reduction potentials follow similar patterns.


Chromosomal breakage in Crohn's disease: anticlastogenic effect of D-penicillamine and L-cysteine. The incidence of chromosome breakage was found to be elevated in 42 patients with Crohn's disease. This phenomenon was much more striking in cultures set up with TCM 199 than in cultures set up with RPMI 1629 rich in L-cysteine. The drug D-penicillamine, a close analog of L-cysteine, gave an apparent therapeutic response in several patients and reduced the chromosome breakage frequency in the lymphocytes of these patients in vitro and in vivo.


Subclinical effects of chronic increased lead absorption--a prospective study. I. Study design and analysis of symptoms. Seventy workers exposed to lead for at least one year and 35 control workers have been enrolled in a prospective study of possible neurologic effects of chronic lead absorption at or below the current standard of 80 microgram per 100 ml whole blood. The study design is described in detail. Initial results of analysis of lead-related symptoms from baseline studies indicate few differences between the exposed and nonexposed workers. The majority of differences were for central nervous system (CNS) symptoms and muscle or joint pain. Little correlation was found between symptom reporting and indices of lead absorption. The evidence suggests that factors other than lead absorption itself may be important in symptom reporting.


[Treatment of cardiac insufficiency in the acute stage of cardiac infarct]. The studies on Strophantin effect upon the hemodynamics in the acute stage of cardiac infarction has revealed that it elevates the pressure in pulmonary artery and reduces the cardiac output, in certain cases deteriorating the patient's state. Furanthril and Droperidol reduce the cardiac output but at the same time diminish the pressure in the pulmonary artery and reduce the pulmonary resistance, thus having a favourable effect on the patient's state. Proceeding from the results obtained, a treatment scheme, including Furanthril, Droperidol and Strophantin, is recommended for the cardiac insufficiency treatment in the acute stage of cardiac infarction depending on the disease onset and the degree of cardiac insufficiency manifested.


[Dynamic changes in the indices of myocardial infarct patients at the polyclinical stage of rehabilitation]. Twenty five patients with myocardial infarction were followed up under the conditions of systematic rehabilitation. They were tested by the veloergometer at the beginning of the fourth and seventh month post the coronary incident. The dynamics of the indices concerning the outer work and oxygen consumption of the myocardium was followed up. The author found that the indices of outer work (capacity of threshold loading and volume of the work carried out) reflect the improved physical capacity of the patients respectively. The dynamics of the indices for the heart work, myocardial oxygen consumption respis not so regular and puts certain problems forward. That fact demanded the inclusion of additional parameters, being revelant to the myocardial contractility. In this way, the rehabilitation regimen could be more precisely determined as well as the effectivity of the therapeutic measures carried out.


[Circulation immune complexes in chronic glomerulonephritis]. Thirty six patients with chronic glomerulonephritis, 35 -- with chronic pyelonephritis, 5 -- with diffuse glomerulonephritis with systemic lupus erythematosus and 60 healthy subjects were examined with the method by sedimentation with 3.5 per cent solution of polyethyleneglycol for the determination of circulating immune complexes. The average value for the healthy subjects is X = 0.123 +/- 0.047 mg/ml. The average value plus two standard deviations = 0.217 mg/ml is accepted as normal in healthy subjects. An elevated level of circulating immune complexes is found in 60 per cent of the patients with lupus nephritis and 25 per cent of the patients with chronic glomerulonephritis. The average values for the last two groups are X = 0.475 +/- 0.554 mg/ml and X = 0.184 +/- 0.185 mg/ml respectively. Whereas in the patients with lupus nephritis the values for the single patients are considerably over the adopted norm, in patients with chronic glomerulonephritis, in the majority of the cases, they are about its upper limit.


[Abdominal manifestations of chronic pyelonephritis]. The most frequent abdominal complaints in chronic pyelonephritis are analyzed and one patient is described, who with the exacerbation of the disease manifested the clinical picture of acute abdomen. The involvement of visceral-dermal reflexes with hyperalgesia, muscular defense, vomiting, meteroism and disturbances in intestinal motility imitated the picture of acute abdominal drama, forcing the surgical intervention. The varied abdominal symptomatics in about 20 per cent of the cases with chronic exacerbated pyelonephritis, should be critically assessed by the therapeutists.


[Echocardiographic indices of the pumping and contractile functions of the left ventricle and the hemodynamic data obtained from left and/or right cardiac catheterization]. A juxtaposition of the EchoC-indices for the pump and contraction function of left ventricle and the hemodynamic data obtained with right and left cardiac catheterization was carried out in 95 patients with different myocardial and valvular heart diseases. The following EchoC-indices were determined for the assessment of the left-ventricle function -- telediastolic dimension and volume of left ventricle, stroke and minute heart volume, fraction of shortening (FS), average contraction speed of circumferential fibres (CCF), fraction of expulsion (FE), left auricle dimension. A considerable correlation was found among the EchoC-indices for pump and contraction function of left ventricle and the final diastolic left ventricle pressure and the average left auricle pressure. In heart patients with a normal final diastolic pressure in left ventricle and average left auricle pressure, pulmonary capillaries (PC) resp., the average values of all functional EchoC-indices are normal. With an elevated final diastolic pressure in left ventricle, diminished values of dP/dt max and elevated mean pressure in left auricle, the pulmonary capillaries resp., the values of all EchoC-indices for the pump and contraction left ventricle function are pathological -- enlarged telediastolic dimension and left ventricle volume, enlarged dimension of left ventricle, reduced values of FS, CCF, FE, SV and MV. EchoC-functional indices properly differentiate the pathological from the normal hemodynamics of left ventricle. determined by catheterization in 86 per cent of the cases. The false positive and false negative results are obtained in 14 per cent of the cases.


[Bicuspid aortic valve. Echocardiographic diagnostic potentials]. The echocardiographic potentialities in the diagnosis of bicuspid aortic valve are discussed. In the group of 32 patients examined with a clinic and murmurs suspected for that disease, the index of excentricity in five patients, was found to be respectively 1.6, 1.4, 1.6, 1.8, 1.5, accepted as a reliable index for the diagnosis of aortic bicuspidia. Excentricity index is equal to 1/2 of the aortic diameter, divided to the minimal distance from the diastolic echo of the closed aortic valve to the closer aortic wall. The control group consisted of 44 patients with tricuspid aortic valve, 17 of them being with aortic defect (isolated stenosis or combined aortic defect) and the rest 28 -- without valvular heart disease. Excentricity index for the control group was low -- 1.0--1.33 (average value 1.28). Conclusions are drawn that the echocardiographic method, from the group of the bloodless methods, is with the best diagnostic potentialities in the bicuspid valve and should be looked for in patients with expulsion murmur and click of aortic origin.


[Ischemic heart disease and embolisms and thomboses in the lesser circulation]. All deceased with ischemic heart disease (IHD) -- 450 cases and with embolism and thrombosis in pulmonary circulation (ETPC) -- 601 cases were purposefully examined for a period of 10 years (1963--1972). In 75 of the examined a combination of IHD and ETPC was present. In 41 out of them only cicatrices of past infarctions were found and in 34 -- recent myocardial infarctions. In 14 of the examined (41%) it was established, clinically and morphologically, that ETPC are with a longer duration than the recent myocardial infarctions and very likely have played a role in their pathogenesis. The confirmation of acute ischemic changes in myocardium gives a support -- undulation of the myofibrils and fuchsinophylic necrosis in the deceased of ETPC without preceeding IHD. The cardiac hypertrophy and coronary atherosclerosis lead to an intensification of the myocardial ischemic alterations in ETPC.


[Excretion of uric acid in cardiac decompensation]. The role of hyperuricemia is confirmed, in literature, as a risk factor in ischemic heart disease and in the development of atherosclerotic complications. The problem of uric acid concentration in serum is still not elucidated as well as the role of the eventual hyperuricemia as a risk factor and its excretion from the kidneys of patients with heart diseases in a decompensation stage. Thirty five patients with different heart diseases in various decompensation stages were observed. In all patients, the uric acid concentration in serum were studied as well as the clearance of uric acid, reabsorption percentage in tubules and glomerular filtration according to the clearance of endogenous creatinine. The results obtained revealed that hyperuricemia (over 7 mg%) is found in a great number of the patients (40%) with heart decompensation. The hyperuricemia established resulted primarily from the reduced filtration of glomeruli. In some of the cases, an increased reabsorption of uric acid in tubules was found. In other, single cases, data exist about uric acid secretion in tubules. The problem of heart patients treatment in decompensation stage with the presence of hyperuricemia is discussed. In those cases, very likely, prophylactic measures should be undertaken due to the fact that hyperuricemia is a factor, secondarily leading to atherosclerotic heart alterations.


[Malignant, rapidly progressing nephritis in Schönlein-Henoch disease]. A case of malignant quickly progressing Henoch-nephritis is described, with a lethal end one and a half month after the initial manifestations. The disease develops with a clinical picture typical for systemic vasculitis, with joint-pain, skin-rash and grave abdominal syndrome, being the reason for unnecessary surgical intervention. The renal changes originate even at the very beginning of the disease and are clinically characterized, besides the exceptionally quick evolution, by the picture of acute renal insufficiency with progressing azotemia, severe electrolyte disturbances and anemic syndrome and pathomorphologically by the picture of severe diffuse intra- and extracapillary proliferative glomerulitis. The treatment with antihistamine preparations, heparin and corticosteroids failed to give an effect.


[Particular ultrastructural aspects of human Paneth's cells]. Our research is about the ultrastructural aspects of Paneth cells obtained from human jejunal mucosa. The cells appear columnar in shape and show a roundish nucleus; their cytoplasm is characterized by a great amount of R.E.R. and by typical large granules in sopranuclear position. Our results confirm those previously described by other authors about the morphology of Paneth cells. In our specimens we have found moreover a variable number of particular cytoplasmic structures; these may show a fibrillar or granular matrix and are always surrounded by a membrane.


[Prolactin after repeated intravenous injections of cimetidine]. To 4 normal women a bolus of mg 400 of cimetidine was injected intravenously once an hour for three hours in succession and plasma prolactin behaviour studied. No depletion in prolactin pituitary storage was observed in all subjects. The possible usefulness of cimetidine for clinical diagnostic purposes is discussed.


A multi-channel, tele-metric system for analysing biological data: extern work, the electrocardiogram, etc... A new multichannel, telemetric system for analysing biological data was developed and tested, in collaboration with the Berta Firm in Turin. This system was tested on subjects at 4650 meters in Peru. The results obtained show the usefulness of this tool. In addition it is very good in evaluating the progress of post-myocardial infarctuated patients.


Thoracic and pulmonary radiographic modification during immersion in apnea: 1--a special apparatus. Thoracic and pulmonary radiographic modification during immersion in apnea, were studied. An Apparatus "Nuclearisub" was used, it consists of two parts: An x-ray generator (100 KV, 40 mA) and a image intensifier tube (brightness gain 1200) with a transistorized telecamera. Through continuous radioscopy during immersion, can be analyzed the variations caused by redistribution of the blood, as well as the changes in pressure.


[Experimental poisoning with gaseous hydrocarbons: changes in concentrations of propane and butane in the lung and adipose tissue in relation to the time of death]. Postmortal changes of propane, butane and iso-butane concentration in the lung and fat of mice exposed to lethal intoxication with commercial liquid gas are studied. The hydrocarbon tissue concentrations after exposure were determined by gas liquid chromatography. The hydrocarbons progressively decreased in the lung and disappeared, depending on the degree of putrefaction; even in case of remarkable putrefaction, it was still possible to detect them in the fat tissue.


Effect of acute arterio-venous fistulae on mean and phasic coronary flow in the dog. The effect of acute arterio-venous fistula on mean and phasic coronary flow was studied on 4 open-chest dogs. When the pressure is basically high, after the opening of the fistula the fall which coronary flow shows during the isometric systole occurs to a lesser extent; conversely, when the pressure is basically normal, the fall of the coronary flow may be absent.


[Cholinesterase of the chick optic lobe. Molecular forms and behavior after deafferentation]. The behaviour of AChE and ChE has been studied quantitatively in the chick optic lobe (tectum and nuclei) under normal conditions and after deafferentation. Eye extirpation was carried out at the 1st day after hatching. Both in the tectum and in the nuclei, a low decrease in specific activity was observed, in comparison with the control. Nevertheless, when the total activity is calculated, a significant decrease is evident, owing to marked underdevelopment of the deafferented lobe. In the normal chick lobe, 2 molecular forms of AChE (6.58 and 11S) have been observed. The distribution of these 2 forms is not altered by eye extirpation, until at least 120 days of age.


Evidence for heterogeneous turnover of rat liver lysosome hydrolases. The activity of several Lysosome Hydrolases in the liver of rats submitted to intensive treatment with cycloheximide has been tested. From their results the authors suggest that the lysosomal turnover is not only heterogenous but also rapid.


Inhibition of liver protein degradation in vivo by intensive treatment with cycloheximide. The AA in experiments performed on male rats treated with 0,5 mCi/kg of cycloheximide have observed an inhibition of liver protein degradation and have suggested two mechanism: 1) cycloheximide prevents the acute proteolytic response induced in fibroblast culture by exposure to serum-deficient media and in rat livers after perfusion probably by inhibiting the lysosomal - autophagic system, without modifications in total activity of lysosome proteinases. 2) It may be that protein degradation rate and level of lysosomal proteinases reinterrelated; the latter probably being a factor controlling the overall cell protein turnover rate.


Fluctuations in the level of liver cathepsins after partial hepatectomy. Fluctuations in the level of liver cathepsins after partial hepatectomy were studied. The reduction in proteinase activities in dividing cells might conceivably reveal larger than that measured on the whole tissue since no more than 40-50% of residual liver cells takes part into the first mitotic wave after partial hepatectomy.


Acetylcholine's inhibitory effect on the calcium-dependent depolarization in the heart cells of a reptile (Lacerta sicula campestris). Preliminary notes. The results of experiments performed with high - K+ solutions suggest that in the lizard atrial cells Ach normally displays its negative inotropic effects by a double mechanism = an "indirect" inhibition, mediated by the increase of the membrane permeability to K+ ions; and a "direct" inhibition of the Ca- carried slow inward current. In the ventricular fibres there isn't a direct-effect of this drug, under normal conditions.


Uncontrolled ventricular rate in atrial fibrillation. A manifestation of dissimilar atrial rhythms. A patient with coarse atrial fibrillation and a rapid ventricular response developed periods of high grade atrioventricular block interpersed with periods of rapid ventricular conduction after the administration of digitalis and propranolol. Intracardiac atrial recordings showed similar atrial rhythms of high right atrial flutter and left atrial fibrillation. The low right atrial recordings showed flutter during the periods of fast ventricular rates and fibrillation during periods of slower ventricular rates.


Coronary artery spasm of different degrees as cause of angina at rest with ST segment depression and elevation. Chest pain induced by ergometrine was associated with ST depression and subsequent elevation in anterior leads in a 42-year-old man suffering from angina at rest. Coronary arteriography during the attack showed that coronary arterial spasm of different degrees was responsible for chest pain and both types of electrocardiographic abnormality.


Echocardiographic diagnosis of mobile left ventricular tumour. Echocardiography detected a mobile left ventricular tumour. The tracing showed a cluster of echoes in the left ventricular outflow tract corresponding to the location of the tumour as confirmed by cineangiograms. At operation, a thrombus was discovered, attached to a chorda tendineae.


Double outlet from chambers of left ventricular morphology. This series of 5 cases with double outlet of morphologically left ventricular chamber includes 4 found during a review of 1700 heart specimens (incidence 0.23%) and 1 found at operation and successfully corrected. Abnormal atrioventricular connection precluding total correction was present in the 4 anatomical cases. Clinical diagnosis may be difficult and it is suggested that axial cineangiography may make anatomical diagnosis easier. Absence of the infundibular septum and aortic laevoposition are frequent. As some cases can be surgically corrected, accurate information is required on the size of the right ventricle, the morphology and function of the atrioventricular valves, the presence, size, and position of the ventricular septal defect, and the degree and type of outflow tract obstruction.


Two-chambered right ventricle: simulating two-chambered left ventricle. Two cases are described of a most unusual variant of two-chambered right ventricle. In both the ventricular septal defect was between the distal chamber of the right ventricle and the left ventricle. However the extensive dividing 'septum' between proximal and distal parts of the right ventricle converted the latter, haemodynamically, into part of the left ventricle. In the first case the distal chamber supported the aorta in the left anterior position, the pulmonary artery arising from the proximal part of the right ventricle. In the second the pulmonary artery arose from the distal chamber and the aorta from the proximal chamber. Though in both the ventriculoarterial connection was double outlet right ventricle, functionally there was arterial concordance in case 1 and discordance in case 2. A further disconcerting feature was the resemblance of the distal right ventricular chamber to the rudimentary chamber of a univentricular heart of left ventricular type.


Haemodynamic effects of dobutamine with special reference to myocardial blood flow. A comparison with dopamine and isoprenaline. The haemodynamic effects of dobutamine (2.5 to 10 micrograms/min per kg) were determined in 5 patients without cardiac failure who were undergoing cardiac catheterisation for suspected coronary disease. Myocardial blood flow was determined by the coronary sinus thermodilution technique. Data were compared with those from two groups of 5 patients who received dopamine (4-8 micrograms/min per kg) and isoprenaline (0.005-0.025 micrograms/min per kg). Each drug was given in a lower and a higher dose, and all increased mean cardiac index (dobutamine, 18% and 39%; dopamine, 11% and 23%; isoprenaline, 15% and 44%). These increases were associated with significant increases in mean myocardial oxygen consumption (dobutamine, 38% and 61%; dopamine, 25% and 62%; isoprenaline, 20% and 45%). Mean myocardial blood flow was increased by each drug but mean myocardial oxygen extraction was decreased by isoprenaline, was increased by dopamine, and was unchanged by dobutamine. Each inotropic agent has a similar effect on myocardial oxygen consumption, but isoprenaline has a direct coronary vasodilator action while dopamine has a coronary vasoconstrictor action. Dobutamine has no direct effect upon coronary vascular tone.


Age, sex, and ischaemic heart disease as prognostic indicators in long-term cardiac pacing. The influence of age, sex, and overt ischaemic heart disease on survival after pacing was studied in a population of 427 patients. The median survival for the whole group was 7.75 years. Women between 55 and 74 years and men between 65 and 74 years enjoyed median survivals in excess of 10 years. Patients over 74 years and men with ischaemic heart disease had median survivals of the order of 5 years and 3 1/2 years, respectively. The findings may help to match generator lifetimes to patient-survival prospects.


Persistence and effects of sinus rhythm after Fontan procedure for tricuspid atresia. Four patients who had had a Fontan type of procedure for tricuspid atresia 23, 6, 6, and 11 months previously were investigated by ambulatory electrocardiographic recording and simultaneous recording of the jugular venous pressure and echocardiogram of the conduit or pulmonary valve. All had been considerably improved by the operation. In 1 patient episodes of supraventricular tachycardia were recorded but no rhythm disturbance was detected in the other 3. Pulmonary blood flow was shown to be pulsatile and atrial systole is an important factor in this. The conduit valve showed delayed opening and slow closure suggesting that its presence in the pulmonary circuit may be unnecessary.


Prinzmetal's angina:reflex cardiovascular response during episode of pain. Previous angiographic studies have shown that coronary spasm occurs in association with the variant angina described by Prinzmetal, confirming his original hypothesis. In this work we recorded the heart rate changes and the incidence of arrhythmias during variant angina. The patients were divided into two groups: anterior, with electrocardiographic signs of anterior ischaemia, and inferior, with changes in the inferior leads. There was a significant increase of heart rate during pain in anterior myocardial ischaemia and a significant decrease when the ischaemia was inferior. The incidence of ectopic arrhythmias during pain was significantly greater in patients with anterior ischaemia, but there was a high incidence of atrioventricular block in patients with inferior ischaemia. We suggest that these findings can be explained by different responses of the automatic nervous system to anterior and inferior acute myocardial ischaemia.


Atresia of the right atrioventricular orifice with atrioventricular concordance. Three hearts are described in which a fibrous membrane was interposed between the right atrium and a formed but hypoplastic right ventricle, which possessed recognisable inlet, trabecular, and infundibular components. In these hearts the distribution of the conducting tissue was as expected for concordant atrioventricular connections, and contrasts with that seen in the classical type of 'tricuspid atresia'. The distinctive morphological and histological features of these specimens lend further support to our view that the majority of cases of atresia of the right atrioventricular orifice should be regarded as coming within the designation of 'the univentricular heart'.


Truncus arteriosus communis with intact ventricular septum. This is the first documented case of truncus arteriosus communis with intact ventricular septum in which extensive clinical, haemodynamic, angiographic, and pathological data are available. Angiography suggested the presence of two discrete semilunar valves but necropsy showed a basically single semilunar valve. This case fills a gap in the spectrum of aorticopulmonary, truncal, and infundibular septal defects, and reinforces the belief that the essence of truncus arteriosus communis is a single semilunar valve, common to both ventricles, which need not be associated with the defects in the adjacent parts of the structural continuum.


Drugs and human memory: effects of low doses of nitrazepam and hyoscine on retention. 1 The effects of diazepam (5 mg) and hyoscine hydrobromide (0.3 mg) were assessed in two memory tasks: short-term retention of digit strings and the free recall of items from categorizable lists. 2 One hundred and two healthy subjects were tested in an independent-groups design. Subjects were assigned randomly to either placebo, diazepam or hyoscine groups. Treatments were administered orally under double-blind conditions. 3 The short-term retention of digits was impaired to an equivalent degree and locus for both drugs (P less than 0.05). This effect was ascribed to the action on primary memory. 4 The drugs produced no significant effects on the recall of categorizable lists either in terms of the number of words recalled or the cohesiveness of categorical recall. 5 These results demonstrate that drugs of different pharmacological actions produce isomorphic psychological deficits in memory and that 'anti-memory' effects on one task should not be extrapolated to all aspects of memory.


The activity of aryl hydrocarbon hydroxylase in adult human skin. 1 Aryl hydrocarbon hydroxylase (AHH), a mixed-function oxidase system, has been identified in microsomal preparations of adult breast skin and foreskin. 2 Separation of dermis from epidermis by stretching, showed that AHH activity in human skin is almost exclusively located within the epidermis. 3 Preincubation of whole chopped skin with benzanthracene (5 muM to 100 muM) using a tissue culture system was accompanied by a concentration dependent increase in AHH activity. 4 Although there was no significant difference in AHH activity between the two sites, individuals differed markedly from one another in activity at any one site. Activity was observed to be positively correlated with age.


Phenytoin absorption in patients with ileojejunal bypass. 1 The absorption and elimination of phenytoin was studied after a 200 mg oral dose was administered to seven patients with ileojejunal bypass and nine control patients. 2 Analysis of the area under the plasma concentration time curve showed the absorption of phenytoin was decreased in the subjects with ileojejunal bypass (P less than 0.005). 3 The half life of elimination of the drug was shorter in the bypass group (15.1 h) than in the controls (17.8 h, P less than 0.05). 4 Subjects with an ileojejunal bypass receiving phenytoin are likely to require increased oral dosages to achieve an optimal plasma concentration. There is probably malabsorption of other poorly soluble drugs and parenteral therapy may be necessary to ensure adequate bioavailability.


The effect of food and tablet formulation on plasma prednisolone levels following administration of enteric-coated tablets. 1 Plasma prednisolone levels have been compared following the administration of enteric-coated prednisolone to fasted and non-fasted subjects. The effect on plasma levels of altering the formulation of the enteric-coating has also been studied. 2 The presence of food in the stomach at the time of administration does not affect the absorption of enteric-coated prednisolone tablets. 3 There was considerable inter-subject variation in plasma prednisolone levels after administration of shellac based enteric-coated tablets. However, plasma levels were more consistent when a preparation whose formulation was based upon cellulose acetate phthalate (CAP) was given. 4 It is concluded that the pattern of absorption and plasma prednisolone levels depend on the formulation of the enteric coating. The bioavailability of the CAP based preparation is similar to that of plain prednisolone.


Tumour site and renal dysfunction as factors influencing leucopenia after chemotherapy for Burkitt's lymphoma. Forty-four (44) patients with Burkitt's lymphoma received identical combination chemotherapy on the basis of body surface area. Patients with renal dysfunction, more common in those with abdominal tumours, were at significantly greater risk of developing severe leucopenia (less than 1000 cells/dl) than those with normal renal function (P less than 0.0001). Similar results were seen in a series of 8 patients with normal marrows treated with only i.v. cyclophosphamide and intrathecal methotrexate. Giving a lower initial dose of cyclophosphamide seemed to reduce the risk of severe leucopenia in 5 additional patients with evidence of renal dysfunction. The mechanism is postulated as delayed excretion of the active metabolites of cyclophosphamide. Adjustment of the chemotherapeutic dose should be considered when treating patients with renal dysfunction.


Tissue distribution and tumour localization of 99m-technetium-labelled liposomes in cancer patients. The possible use of liposomes (phospholipid vesicles) to direct cytotoxic drugs to tumours has led us to investigate the tissue localization of i.v. injected 99m-Tc-labelled liposomes in cancer patients. Twenty mg or 300 mg doses of liposomal lipid (7:2:1 molar ratio of phosphatidylcholine : cholesterol : phosphatidic acid) were used in a study of 13 patients with advanced cancer and one with polycythaemia rubra vera (PRV). In all cases except the patient with PRV the major site of uptake of the label was the liver and spleen. In the patient with PRV the liver uptake was greatly reduced and the major site of uptake was found in regions corresponding to marrow. With the exception of one patient with a primary hepatoma, there was no significant tumour uptake of the label.


Results of treatment of female breast cancer in the Cambridge area 1960-71. The results of treatment of female breast cancer in the Cambridge area during the period of 1960-71 are presented and compared with an earlier series treated in the period of 1947-50. It is estimated that about 29% of the patients treated in the later period are cured, in the sense that they have a life expectancy similar to that of the normal population, compared with just under 20% in the earlier period, but this improvement is mainly due to an increased proportion of Stage I cases in the latter period. The percentage cured is discussed in relation to the ratio of deaths to registrations in the East Anglian Region and it is suggested that under-registration of deaths from cancer of the breast may occur.


Assays of drug sensitivity for cells from human tumours: in vitro and in vivo tests on a xenografted tumour. A human tumour which grows as a xenograft in immune-suppressed mice and forms colonies in vitro has been used to test the correlation between 2 methods of exposure of human tumour cells to chemotherapeutic agents. In vivo exposure to drugs was achieved by injection of tumour-bearing mice with each of 8 cytotoxic agents. For the in vitro exposure, cell suspensions were incubated for 1 h with the same series of drugs. The survival of tumour clonogenic cells was assayed in vitro after either treatment or dose-response curves were obtained. The 8 drugs were ranked according to their in vivo effect at doses equitoxic to mice, and according to their in vitro effect at concentrations designed to approximate to levels of drugs in human plasma. The ranks for in vivo and in vitro exposure correlated well.


Inhibition of DNA synthesis by nitroheterocycles. I. Correlation with half-wave reduction potential. Twenty-one nitroheterocycles, including metronidazole, misonidazole and AF-2, were tested for their ability to inhibit DNA synthesis in mouse L-929 cells growing in culture. All those tested inhibited the rate of incorporation of 3H-thymidine into L cells following drug treatment for 4 h under aerobic conditions. Only 4 drugs reached their limits of solubility before the uptake of 3H-thymidine was inhibited by 50% or more. For the remaining 17, the log of the concentration producing 50% inhibition of incorporation was directly correlated with the half-wave reduction potential of the compound.


Inhibition of DNA synthesis by nitroheterocycles. II. Mechanisms of cytotoxicity. Nitroheterocycles have been shown to inhibit the incorporation of 3H-TdR by cultured L-929 cells, and the degree of inhibition is related to their electron-affinity. On the basis of their chemical reactivity and potential clinical utility, nitrofurazone, misonidazole and metronidazole were selected for more detailed studies of the mechanism of inhibition of DNA synthesis. Double-isotope labelling in conjunction with hydroxyapatite chromatography allowed the evaluation of drug effects on initiation of DNA replicons, DNA chain elongation and DNA damage (single-strand breaks), and their correlation with eventual cell viability. Partial inhibition of initiation of DNA synthesis generally preceded other measurable effects, and was not reversed by incubation in the absence of drug. In the absence of DNA strand breaks (at low drug doses or after a repair interval) the rate of elongation was similar in both treated and untreated cell populations. Measurable DNA damage (strand breaks) was predictive for cytotoxicity. At lower drug doses, or under aerobic conditions, DNA synthesis was not always associated with a decrease in plating efficiency (cytotoxicity) but was reflected in decreased colony size (growth rate) of the cells. Thus the aerobic "toxicity" previously reported for chronic exposure to these agents may be better described as a "cytostatic" effect. Under anaerobic conditions (where cell killing is much greater) inhibition of initiation plays a less important role, and the nitroheterocycles are metabolically reduced to intermediates which are truly cytotoxic.


Racial differences in melanoma incidence. The incidences of malignant melanoma recorded by 59 population-based cancer registries were investigated to determine the effects of racial and skin-colour differences. White populations exhibited a wide range of melanoma incidences and females commonly, though not invariably, had a higher incidence than males. Non-white populations experienced in general a much lower incidence of melanoma although there was some overlap of white and non-white rates. No predominant sex difference emerged among non-whites. Populations of African descent were found to have a higher incidence than those of Asiatic origin, but it was concluded that this was due largely to the high frequency of tumours among Africans on the sole of the foot. A clear negative correlation between degree of skin pigmentation and melanoma incidence emerged for the exposed body sites. These data provide strong support for the hypotheses that UV radiation is a major cause of malignant melanoma and that melanin pigmentation protects against it. Further research is required to elucidate the aetiology of melanoma of the sole of the foot.


Prognostic significance of modal DNA value and other factors in malignant tumours, based on 1465 cases. The modal DNA values of 1465 tumours, together with other factors of possible prognostic importance, were related to the survival of the patients, using regression models (Kay, 1977). For most tumour sites except the testis, the distributions of modal DNA values were bimodal, with peaks at the diploid level and in the triploid-tetraploid range. For all tumour sites except the cervix uteri, patients in the low (near-diploid) range showed better survival; the reverse was true for squamous-cell carcinoma of the cervix uteri. Other variables showed the following effects: for all sites except the testis, younger patients showed a better survival; for the cervix and corpus uteri, breast and ovary, increasing clinical stage was associated with poorer survival. Where evaluated, histological grade appeared to be associated with survival rate, the less well differentiated tumours having a worse prognosis, except for the breast, where the reverse correlation was noted. For carcinoma of the bladder, females and a poorer survival rate than males.


Giant-cell tumour of bone: cytological studies. Cell-surface markers were investigated in 7 patients with giant-cell tumours and 30 patients with other tumours as controls. 25--55% of mononuclear cells in giant-cell tumours showed immunoglobulin-mediated phagocytosis. These phagocytic cells showed rapid adherence, trypsin resistance and potent nonspecific esterase activity. Thus, giant-cell tumours contained considerable numbers of macrophages with typical characteristics and functions. Macrophages did not proliferate in cultures of giant-cell tumours, whereas the non-adherent cells did. Further, established cell lines from these tumours consisted of spindle-shaped cells without surface markers or the ability to phagocytose or display nonspecific esterase activity. We consider that macrophages, which may be precursors of giant cells in giant-cell tumours, are non-malignant cells of host origin rather than tumour cells acquiring some properties of macrophages. We found that macrophages were more abundant in giant-cell tumours than in other tumours of mesenchymal origin, but any effect of their presence on the clinical behaviour and prognosis of the tumour remains highly speculative.


Inactivation by the mitotic inhibitor NY 3170 of human cells in vitro. Inactivation of NHIK 3025 cells ny the mitotic inhibitor NY 3170 (1-propargyl-5-chloropyrimidin-2-one) was measured as loss of colony-forming ability. NY 3170 at a concentration of 0.15 nM allowed no formation of colonies after 12 days of continuous exposure to the drug. Metaphase arrest after treatment with NY 3170 was reversible if the drug was removed immediately after the onset of the arrest. When the cells were kept in mitosis by the presence of NY 3170, inactivation was complete after 8h incubation of mitotic cells with 0.4 nM NY 3170. Using synchronized cell populations, it was shown that mitosis is by far the most sensitive stage of the cell cycle to inactivation by NY 3170. This leads to the suggestion that there is a connection between the inactivating and the metaphase-arresting effect of this drug. The age response curves show that after mitosis the stages in order of decreasing sensitivity to NY3170 are: G2, late S, early S and G1. This is a similar age response to that reported for proliferating cells treated with bleomycin, whereas the mitotic inhibitors vincristine and vinblastine have shown qhite different age response curves.


Antibody-dependent cellular cytotoxicity to human colon-tumour cells. I. Lack of tumour specificity in a population study. The humoral and cellular components of the antibody-dependent cellular cytotoxicity (ADCC) against allogeneic human colonic tumour cell lines were evaluated. The 2 colon cell lines used in this study (HT-29 and ACC-20) were found by immunofluorescence to have carcinoembryonic antigen (CEA) on their surface, and to become sensitive to the lytic effect of unstimulated lymphocytes after coating with heterologous anti-CEA. This reaction was used to evaluate the ADCC activity of mononuclear cells from the peripheral blood of patients with gastrointestinal cancer (mostly local extensive colo-rectal). Remarkable variability was found in the lytic capability (2-50% specific lysis) of both cancer and non-cancer mononuclear cells, with no significant difference between them. Sera from 127 cancer patients and 91 non-cancer patients were tested, using the reaction with heterologous anti-CEA as positive control and as a reference point. In 46 cases (21%) the sera were reactive in this system, and 43 of them were of Blood Group O. However, there was no difference between the cancer patients and the normal controls. The antigenic determinant involved in this reaction is not the Blood Group A specificity but, most probably, a polypeptide common to CEA and A (as shown in the following publication). In addition, trials for the elimination of the non-tumour-specific reaction, by absorption or inhibition, failed to disclose a tumour-specific one. The value of the ADCC assay in monitoring human tumour immunity, and possible ways of eliminating reactivity to normal antigens in this system, are discussed in the light of these findings.


Tissue distribution and biochemical properties of an interspecific tumour-associated gamma foetal antigen. A late-gestation neonatal antigen (gamma foetal antigen; gamma-FA) immunologically and biochemically unrelated to murine alpha-foetoprotein, was identified in several spontaneous and carcinogen-induced sarcomas and hepatic carcinomas of the mouse and rat. An approximate mol. wt of 35,000 for gamma-FA from both foetus and tumour was obtained by molecular-sieve chromatography and sucrose-gradient centrifugation. Radial immunodiffusion analyses of organ extracts indicated that gamma-FA could be found in several neonatal tissues, the highest concentration occurring in the spleen. In the 2-month-old mouse, only splenic tissue contained gamma-FA and at much lower levels than in the organ of the newborn mouse.


Effects of discontinuation of chronic feeding of diethylnitrosamine on the development of hepatomas in adult rats. Diethylnitrosamine (DENA) at 10 mg/kg was fed to adult rats either continuously or for periods ranging from 1 to 10 weeks. Survival correlated inversely with the duration of carcinogen feeding. Less than 4 weeks of DENA feeding produced only preneoplastic foci that persisted indefinitely; 4 weeks were found to be necessary for the transformation of preneoplastic lesions into liver cancers; after 6 weeks, the incidence of hepatomas was 100%. The process of liver cancerization appeared to be identical whether DENA was fed for 8 weeks or continuously up to the time of death. These results are discussed in the light of the evolution of the homoeostatic control of liver-cell division during DENA feeding, in order to distinguish the different successive roles played by the carcinogen.


Release from the Crabtree effect by hypoxic cell radiosensitizers. The Crabtree effect can be observed when the O2 consumption of tumour cells or of mammalian cells grown in culture is measured in physiological medium containing glucose. The effect of 2 hypoxic cell radiosensitizers, misonidazole and NDPP, on the O2 consumption of Ehrlich ascites tumour cells was compared in media with and without glucose. A stimulatory effect on O2 consumption was found for 5--20mM misonidazole as well as for 0.5mM NDPP, both in media containing 10(-2)M glucose. Thus glucose induced a Crabtree effect in Ehrlich tumour cells, expressed as 38--45% inhibition of O2 consumption relative to that in the same medium without glucose. The stimulatory effect of misonidazole and NDPP on O2 utilization in medium with glucose undoubtedly appeared as a release from the Crabtree effect.


Effect of human cord and postoperative serum on experimental inflammation in the rat. Human cord and postoperative serum depressed the oedema provoked by mediators of the inflammatory reaction such as bradykinin, histamine, serotonin and prostaglandin E2 and also the experimental inflammation caused by carrageenin. Normal human and pregnancy serum did not have such an effect. In two cases of open neural tube defect, one of anencephaly and another of spina bifida, human amniotic fluid also had a strongly depressing effect on the experimental oedema provoked by serotonin. Human amniotic fluid from normal pregnancies did not inhibit this experimental inflammation. A protein-fraction of mol. wt 30,000--100,000 has been isolated from the inhibiting sera and shows the anti-inflammatory activity to be dose-related towards all the oedema-provoking substances used. Immunological studies showed that the inhibiting factor could be a protein in the pre-albuminic region, while alpha-foetoprotein did not appear to be responsible for the anti-inflammatory activity. Our conclusion is that human serum contains a protein of foetal origin with an acute-phase character and strong anti-inflammatory activities analogous to rat alpha 2-macrofoetoprotein.


The effects of perhexiline maleate (Pexid) and alcohol on rat liver. Perhexiline maleate is an anti-anginal drug believed to cause liver damage. The possibility that this effect could be potentiated by alcohol has been investigated. Rats fed both the drug and alcohol did not show more severe liver damage than those fed alcohol alone. Livers from rats fed perhexiline alone did not differ from controls. There is no evidence from this study that alcohol potentiates the effect of perhexiline in rat liver. These results may have a bearing in perhexiline-induced liver damage in humans, but clinical studies are required.


An ultrastructural study of peritoneal mononuclear phagocytes from Corynebacterium parvum-injected mice. Peritoneal mononuclear phagocytes harvested from mice 24 h after i.p. injection of C. parvum displayed hypertrophy of the Golgi and smooth endoplasmic reticulum complex with attendant increase in lysosome production. The ingested bacilli were identified within phagolysosomes, within which large myelin figures accumulated. In addition to heterophagic vacuoles, autophagosomes and lipid droplets were observed. These latter two inclusions may reflect a direct or indirect cytopathic effect of C. parvum on the phagocytes.


Effects of non-haemorrhagic injury on the cardiovascular response to tilting in the rat. In the unanaesthetized uninjured rat a head-up tilt elicited a pressor response, the magnitude of which was directly related to the angle of tilt. The pressor response was reduced by a 4 h period of bilateral hind-limb ischaemia and by a non-fatal scald, although the slope of the regression line relating percentage rise in mean arterial blood pressure to the angle of tilt was unaltered. The degree of impairment of the response was directly related to the severity of the injury. In the uninjured rat there was a sigmoid relationship between the angle of tilt and the mean heart rate during the tilt. No relationship could be demonstrated between angle of tilt and heart rate in the injured rats.