[Familial cardiomyopathy: a study of two families with myocardial and skeletal muscle biopsies]. Cardiomyopathy was diagnosed in several members of two families. This familial cardiomyopathy showed symmetrical or asymmetrical hypertrophy of the ventricular walls with or without obstruction to the left ventricular outflow tract. Certain forms were asymptomatic and were revealed by the family history and echocardiography. Myocardial and intercostal muscle biopsy was performed for a biochemical and ultrastructural analysis. Different myocardial features were observed in the two families: a large increase in the glycogen deposits in the one, without clinical signs of a glycogen storage disease, and intracellular deposits of a filamentous protein substance in the other.


[Fiessinger-Leroy-Reiter syndrome with non-obstructive cardiomyopathy treated with methotrexate]. The case of a 50 year old male with the Fiessinger-Leroy-Reiter syndrome, ankylosing spondylitis and generalised pustular psoriasis is reported. This condition wax complicated by non-obstructive cardiomyopathy, congestive cardiac failure and first-degree atrioventricular block, the site of which was localised by electrophysiological studies (nodal block with an infrahisian conduction defect). After failure of several therapeutic regimes, a spectacular improvement was obtained with Methotrexate associated with a diuretic; the signs of heart failure regressed and the cardiomyopathy stablised. A parallel improvement was seen in the skin, cardiac and articular lesions and has been maintained with an 18 months follow-up. Left ventricular performance was studied by echocardiography. The mechanism of the beneficial effect of Methotrexate is unclear; this therapeutic trial is to be extended to include other cases of primary cardiomyopathy without obstruction.


[Secondary tamponade following cardiac surgery with reference to 3 cases]. Secondary tamponnade after cardiac surgery occurs after a variable period, generally between the 15th day and the 5th post-operative week. Although this is a rare complication, it occurred three times in a consecutive series of 225 patients (1.3 p. 100). Based on this short experience as compared to the number of cases already published, the authors discuss their opinions on: -- the factors of inflammation and post-operative anticoagulation which predispose to this complication; -- the progress in diagnosis brought about by echocardiography; -- the minimal suggested management of pericardocentesis completed or not by surgical drainage; -- the possibilities of prevention based on prolonged follow-up of patients who present post-operative "pericardial problems". In this way it may be possible to eliminate a not negligeable cause of secondary mortality after cardiac surgery.


Separation and identification of the carotenoid pigments of stigmata isolated from light-grown cells of Euglena gracilis strain Z. Thin layer chromatography of the carotenoid pigments of stigmata isolated from light grown cells of Euglena gracilis strain Z resolved 29 compounds, of which 16 could be eluted and their absorption spectra recorded. Seven of these compounds were identified by a combination of co-chromatography with authentic compounds and by chemical tests, one of these compounds (beta-carotene) was further identified by mass spectrometry. The major carotenoids were found to be beta-carotene, diatoxanthin and diadinoxanthin which together comprised approximately 60% of the stigma pigments. In addition significant quantities of canthaxanthin, echinenone and cryptoxanthin were isolated and a possible carotenoid ester was detected. The results of this analysis are compared with those of previous workers and the significance of the findings is discussed.


Modulation of E rosette forming activity of human lymphocytes by rosette inhibiting factor (RIF) ann rosette restoring factor (RRF). The mechanism of action of RIF and RRF on spontaneous rosette formation by human T lymphocytes with SRBC was studied on cells isolated from healthy donors and from patients with active pulmonary tuberculosis. E rosette formation by lymphocytes from healthy donor is inhibited in vitro by RIF. The effect of RIF is reversible since the primary activity of lymphocytes can be restored by contact with RRF. On the other hand, E rosette formation by lymphocytes from tuberculous patients by means of RRF can be depressed by RIF to the starting level. It was found that prostaglandin synthetase inhibitors (indomethacin and RO 20-5720) prevent inhibition or restoration of E rosette activity by RIF and RRF, respectively.


In vitro response to rosette inhibition factor (RIF) and rosette restoring factor (RRF) of human lymphocyte after fractionation on a discontinuous density albumin gradient. In vitro studies on peripheral blood lymphocytes from tuberculous patients and healthy subjects are reported. Factors isolated from human serum used in the experiments included the Rosette Inhibition Factor (RIF) from serum of tuberculous patients and the Rosette Restoring Factor (RRF) from serum of healthy subjects. Lymphocytes were fractionated on a discontinuous BSA gradient. Lymphocytes from healthy subjects, sensitive to RIF, as well as lymphocytes from tuberculous patients, sensitive to RRF, were localized in the third fraction of the BSA gradient. A small percentage of bone marrow cells upon coming into contact with RRF spontaneously formed rosettes with SRBC and were also localized in the third fraction after centrifugation in discontinuous BSA gradient. The results indicate that RIF and RRF act on the same subpopulations, probably precursors of T cells.


Immunogenic properties of slime produced by Pseudomonas aeruginosa. The slime of 8 various Pseudomonas aeruginosa strains was purified and used to immunize rabbits. All studied slime-extracts were strongly immunogenic for animals. The level of antibodies against slime was determined by the passive hemagglutination test. Immune hemagglutinins were present mainly in the IgM fraction of antisera.


Cell fusion induced by invertebrate virus. Brief report. Inoculation of invertebrate cells with Chilo Iridescent Virus (CIV) leads rapidly to massive formation of syncytia. This polykaryocytosis occurs at a high multiplicity of infection and does not require viral genome expression. Thus, cell fusion appears in non permissive conditions, and CIV suspensions rendered non infections by UV irradiation still retain the ability to produce rapid and extensive cell fusion. In the same way, some polykaryocytes could be detected in vertebrate cells (CV1 cell line). However, this process is more effective and faster in invertebrate cell systems.


Synthesis of DNA in cells infected with African swine fever virus. Incorporation of 14C-thymidine by cells infected with African swine fever virus (ASFV) occurs in the nucleus. Part of this DNA is transferred to the cytoplasm and becomes resistant to DNAse. The nuclear fraction washed with Triton X100 retained all labeled DNA and was able to synthesize viral and cellular DNA under in vitro conditions in the presence of the four deoxynucleoside triphosphates, Mg+2, and sucrose. Under similar conditions nuclei from uninfected cells synthesized very little DNA.


Genetic and behavioral studies of female sex appeal in Drosophila. The sex appeal of a Drosophila melanogaster female is defined here as the stimulus (or set of stimuli) which induces wing vibration in courting males. A quantitative measure of sex appeal is the cumulative duration of wing vibration induced by a given female averaged over several consecutive test intervals using different standardized male testers (sex appeal parameter, SAP). By use of SAP, both males and females are found to have the same amount of sex appeal on the first day after eclosion. However, males rapidly lose it by the next day, so that mature males become distinct from females. We report the ontogeny of the male's response to sex appeal. By the SAP method, we also demonstrate that the male's response is dependent on his previous encounter with females. The sex appeal of 287 gynandromorphs was examined in order to localize the sex appeal focus by means of blastoderm fate mapping. Most mosaic flies were classified as either positive (femalelike, with high SAPs) or negative (malelike, with SAPs of zero). Sixteen percent of the gynandromorphs had intermediate levels of SAP, inducing only short vibrations, a response which males rarely give to normal females. Assuming that the gynanders with such intermediate sex appeal must have both female and male foci, distances to the foci from external landmarks were calculated. The center of the focus seems to be an internal structure mapping to the ventroposterior region of the blastoderm fate map, close to the primordia of the anterior sternites. The focus might include a large mesodermal area, but only part of it must have a female genotype for the sex appeal to be expressed. A possible involvement of the fat bodies in production of the sex appeal stimulus is discussed in relation to these findings. Consistent with this conclusion is the fact that females whose abdomens were amputated still retain enough sex appeal to induce male wing vibrations.


Genetic basis of aristal morphology in Drosophila melanogaster and its correlation with behavior: selection for increased and decreased aristal branching. The aristae of Drosophila have been shown to play a role in mating behavior and geotaxis. Two populations of D. melanogaster were selected for increased and decreased numbers of major aristal branches. Selection was successful and resulted in two lines differing by an average of six aristal branches. Hybridization analyses of selected lines revealed that genes influencing aristal branching are located on both the X chromosome and the autosomes. Polygenic control of aristal morphology is indicated by a gradual response to selection and low realized heritabilities. When selection was relaxed for 19 generations, the number of aristal branches did not revert to the number in the control line. Changes in aristal branching did not appear to have a consistent influence on geotaxis, although there was a tendency for flies with fewer aristal branches to be geonegative. Neither mating speed nor ethological isolation between the two populations was affected by selection. It is concluded that the number of aristal branches in Drosophila is a neutral trait (i.e., not subject to natural selection) under laboratory conditions. Correlations between aristal morphology and behavior found in other selection experiments by previous investigators were likely due to linkage disequilibria.


Chemical characterization of a new Japanese variant of carbonic anhydrase I, CA INagasaki 1 (76 arg leads to gln). A new inherited variant of carbonic anhydrase I (CA I), designated CA INagasaki 1 (CA INGS 1), was discovered during a survey of hemolysates from 5852 individuals from the cities of Hiroshima and Nagasaki in Japan. Analysis of the amino acid composition of a tryptic peptide from the CA INGS 1 variant indicated that a glutaminyl residue was substituted for an arginyl residue at position 76. Heat degradation studies showed that the CA INGS 1 variant was less stable than normal CA I. The CO2 hydrase and esterase activities of the normal and variant carbonic anhydrases I, as well as the relative amounts of the two enzymes in heterozygotes, were similar.


An analysis of cytoplasmic RNA populations in Drosophila melanogaster, Oregon R. Nucleic acid reassociation methods were used to estimate the number of different polyadenylated RNA [poly (A) + RNA] sequences in the cytoplasm of whole Drosophila melanogaster at different stages of development and in the cytoplasm of cells of the L3 cell line. The number-average length (LN) of poly (A) + RNA from L3 cells is 1.4 kb, and the steady-state LN of the poly (A) tracts is 70 nucleotides. Analysis of RNA-driven reassociation with copy DNA shows that the poly (A) + RNA from L3 cells contains 5200 different sequences distributed in three abundance classes. The RNA forms hybrid duplexes with about 4.5% of single-copy Drosophila DNA, corresponding to 6500 sequences of 1.4 kb. The LN of poly (A) tracts present in whole embryos, larvae, pupae, and imagos is in each case close to 70 nucleotides. RNA-driven reassociation experiments show that poly (A) + RNA from these sources contains, respectively, 3500, greater than or equal to 4900, 6900, and greater than or equal to 4900 sequences. Cross-hybridization reactions show extensive homology between these RNA populations. All five poly (A) + RNA preparations contain a prominent component with a sedimentation coefficient of 13 and a size of 1.78 kb, which is shown to be the larger species of mitochondrial rRNA. This rRNA binds to oligo (dT) cellulose and serves as an efficient template for cDNA synthesis by reverse transcriptase. In RNA-driven reassociation experiments it behaves like an abundant mRNA.


Increasing the resolution of polyacrylamide gel electrophoresis by varying the degree of gel crosslinking. Resolution of polyacrylamide gel electrophoresis may be substantially improved by taking advantage of the gel sieving effects of varying concentrations of bisacrylamide crosslinker. A dilution procedure is described which permits simultaneous variation of both total acrylamide concentration and percent crosslinking within a single linear regression alaysis.


Purine transport by malpighian tubules of pteridine-deficient eye color mutants of Drosophila melanogaster. Uptakes of guanine into Malpighian tubules of wild-type Drosophila and the eye color mutants white (w), brown (bw), and pink-peach (pp) have been compared. Tubules for each of these mutants are unable to concentrate guanine intracellularly. The transport of xanthine and riboflavin is also deficient in w tubules. The transport of guanosine, adenine, hypoxanthine, and guanosine monophosphate is similar in wild-type and white Malpighian tubules. These data and other information about these mutants make it likely that these pteridine-deficient eye color mutants do not produce pigments because of the inability to transport a pteridine precursor. This view supports the hypothesis that mutants which lack both pteridine and ommochromes do so because precursors to both classes of pigments share a common transport system.


Properties of allophycocyanin II and its alpha- and beta-subunits from the thermophilic blue--green alga Mastigocladus laminosus. Purified allophycocyanin II and its subunits have been examined with respect to spectroscopic properties, sedimentation, reconstitution and isoelectric behaviour. In 0.02m-potassium phosphate buffer, pH8.0, and at a concentration of 0.25mg/ml, allophycocyanin II and its alpha- and beta-subunits show visible absorption maxima at 650, 615 and 615nm respectively, whereas the fluorescence emission maxima were determined to be at 662, 640 and 630nm respectively. The absorption difference spectrum (dilution difference) of allophycocyanin II displays maxima at 650 and 590nm with a minimum at 610nm. The c.d. spectrum of allophycocyanin II showed only one positive-ellipticity band at 635nm, and a major negative-ellipticity band at 340nm. Oxidation of allophycocyanin II, low- and high-pH solutions (pH3.0 and 11.0), various ethanol concentrations as well as dialysis against distilled water induce a spectral change leading to phycocyanin-like characteristics. In most cases these shifts are reversible. Allophycocyanin II is thermostable over a period of 60min at temperatures up to 60 degrees C. The isoelectric points of allophycocyanin II and its alpha- and beta-subunits are 4.65, 4.64 and 4.82 respectively. Estimated molecular weights from sedimentation-equilibrium analyses were 102500 for allophycocycanin II, 16000 for the alpha- and 31500 for the beta-subunit. Recombination of alpha- and beta-subunits leads to allophycocyanin II, which is indistinguishable from native allophycocyanin with respect to its spectral form, to its gel-filtration and to its electrophoretic behaviour.


Purification and characterization of NADPH--cytochrome c reductase from the midgut of the southern armyworm (Spodoptera eridania). 1. NADPH-cytochrome c reductase was solubilized with bromelain and purified about 400-fold from sucrose/pyrophosphate-washed microsomal fractions from southern armyworm (Spodoptera eridania) larval midguts. 2. The enzyme has a mol.wt. of 70 035 +/- 1300 and contained 2 mol of flavin/mol of enzyme consisting of almost equimolar amounts of FMN and FAD. 3. Aerobic titration of the enzyme with NADPH caused the formation of a stable half-reduced state at 0.5 mol of NADPH/mol of flavin. 4. Kinetic analysis showed that the reduction of cytochrome c proceeded by a Bi Bi Ping Pong mechanism. 5. Apparent Km values for NADPH and cytochrome c and Ki values for NADP+ and 2'-AMP were considerably higher for the insect reductase than for the mammalian liver enzyme. 6. These are discussed in relation to possible differences in the active sites of the enzymes.


Characterization and localization of human placental ferritin. Ferritin has been purified from normal full-term human placentae and its antigenic and molecular characteristics compared with adult liver ferritin. Placental ferritin is composed predominantly of a single subunit type, co-migrating with a liver ferritin standard on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Comparison of dose-response curves in an immunoradiometric assay indicated some tissue-specific antigenicity for placental ferritin. This was supported by immunofluorescence studies on cryostat sections of human placentae by using antibodies to placental and spleen ferritin. Specific staining for placental ferritin was demonstrated within placental syncytiotrophoblast, particularly localized towards the microvillus plasma membrane. Ferritin has also been shown by electrophoretic and antigenic analysis to be present in protein fractions solubilized from isolated human syncytiotrophoblast microvillus plasma-membrane preparations, suggesting that ferritin may play an active role in the transfer of iron from maternal transferrin across the syncytiotrophoblast plasma membrane.


Stimulation by secretin of bilirubin UDP-glycosyltransferase activities and of cytochrome P-450 concentration in rat liver. The activity of bilirubin UDP-glucuronyltransferase in liver tissue was increased 1.5-fold after 90 min of secretion administration (4 i.u./h per kg body wt.) in anaesthetized Wistar rats biopsied half-hourly over a period of 2 h. In unanaesthetized R/A Wistar rats, activities of liver enzymes were assayed after administration secretin for 1 h. Bilirubin UDP-glycosyltransferase activities and cytochrome P-450 concentration were increased, but p-nitrophenol UDP-glucuronyltransferase and UDP-glucose dehydrogenase activities remained unchanged.


Gold-induced changes in the morphology and functional capabilities of human monocytes. The capacity of gold compounds to induce morphologic changes and alterations in the functional activity of human mononuclear phagocytes (M phi) in vitro was examined. Human peripheral blood mononuclear cells were incubated with gold sodium thiomalate (25 microgram/ml) for 96 hours. As a result, M phi developed electron dense precipitates within phagolysosomes, as well as marked dilatation of these organelles. Gold incubation also altered a number of M phi functions. While viability and adherence were unaffected, the capacity to spread on surfaces was diminished. Pinocytosis of soluble proteins and phagocytosis of opsonized sheep erythrocytes were impaired, but Fc mediated particle binding was not. These data indicate that gold can alter certain functional activities of M phi and support the idea that the major action of gold in rheumatoid arthritis results from its capacity to alter M phi function.


The gastric antisecretory activity of 3-methyl-5,6,7,8-tetrahydroquinoline-8-thiocarboxamide hydrochloride (tiquinamide). 3-Methyl-5,6,7,8-tetrahydroquinoline-8-thiocarboxamide hydrochloride (tiquinamide) possesses potent gastric antisecretory activity in a number of animal species. It causes a marked inhibition in the volume and concentration of basal and chemically-stimulated gastric acid secretion in the conscious rat and guinea-pig, and of chemically-stimulated secretion in the anaesthetised cat, dog and monkey. Marked activity is also seen in conscious Heidenhain-pouch dogs, against secretion stimulated by betazole and gastrin tetrapeptide. In the pylorus-ligated rat preparation, the antisecretory effect of 30 mg/kg tiquinamide persists for 8-9 h. The possible mode of action of this drug, in view of its wide non-selective antisecretory profile, is discussed.


beta-lactoglobulins in the mammary secretions of camel (Camelus dromedarius) and she-ass. Immunological detection and preliminary physico-chemical characterization. The mammary secretions of a monogastric, the ass, and those of a Tylopode, the camel (camelus dromedarius), were examined by double diffusion in agarose gel against rabbit sera anti-bovine beta-lactoglobulin. Clear precipitin reactions were obtained. After immunoelectrophoresis the camel and she-ass beta-lactoglobulins showed very different electrophoretic mobilities.


[Collagen and hydroxyproline in carcass and skin of young rats fed with hypoproteic and hypocaloric diet]. The AA, reports the variations of collagen of carcass and skin of overnourished and proteic and caloric malnourished rats. The level of collagen is higher in malnourished when compared with normal and overnourished rats. The insoluble hydroxyproline concentration is higher in malnourished than in overnourished rats. This is reported to a variation of metabolic enzymatic patterns related to the catabolism of collagen.


Whole body potassium and total exchangeable potassium in elderly patients with cardiac-failure. Body potassium status of patients with cardiac failure may be estimated by a number of methods, but increasing reliance is being placed upon radioisotope dilution with 42K which measures the total exchangeable potassium. Total exchangeable potassium comprises between 86 per cent and 97 per cent of whole body potassium in healthy subjects. We have measured total exchangeable potassium in 22 oedema-free elderly patients with stable cardiac failure and compared the results with simultaneously determined measurements of whole body potassium obtained by whole body counting. The mean whole body potassium was 2360 +/- 640 mmol. The mean value of total exchangeable potassium measured at 24 hours was 1820 +/- 610 mmol (77% of whole body potassium) and increased further to 2000 +/- 600 mmol (84%) when measured after 48 hours. In patients with cardiac failure and, perhaps, also other patients with a history of fluid retention, the mixing of a tracer dose may be significantly delayed, which if not appreciated may lead to an overestimate of potassium depletion.


Haemodynamic effects of dopamine and dopamine combined with nitroglycerin in patients subjected to coronary bypass surgery. The haemodynamic effects of dopamine and dopamine with nitroglycerin were evaluated in eight patients with coronary heart disease who underwent aortocoronary bypass surgery. The study was performed under anaesthesia and before surgery. Dopamine 8 micrograms kg-1 min-1 alone produced a marked increase of the cardiac index from 2.47 to 3.47 litre min-1 m-2 but only small changes in heart rate (from 65 to 68 beat min-1). This improvement in cardiac performance was accompanied by an increase of the mean pulmonary artery pressure from 10.9 to 21.3 mm Hg and in the left ventricular filling pressure from 6.1 to 13.8 mm Hg with unchanged systemic and pulmonary vascular resistance. Mean arterial pressure increased from 72 to 103 mm Hg. Simultaneous infusion of dopamine (8 micrograms kg-1 min-1) and nitroglycerin (mean dose 0.5 microgram 21.3 to 14.4 mm Hg) and of left ventricular filling pressure (from 13.8 to 7.9 mm Hg). Cardiac index (from 3.47 to 3.34 litre min-1 m-2) and mean arterial pressure (from 103 to 95 mm Hg) were not reduced to the same extent by the addition of nitroglycerin. The combined treatment of dopamine with nitroglycerine seems to be of value in patients with pre-existing high left ventricular filling pressure or with pulmonary hypertension.


65Zinc absorption in patients suffering from acrodermatitis enteropathica and in normal adults assessed by whole-body counting technique. 65Zinc absorption was studied in five acrodermatitis enteropathica (AE) patients and in eight normal adults by means of a whole-body counting assay. The absorption was calculated from retention values recorded in the time interval 8-30 days after oral administration of the isotope. Two AE patients (7 and 13 years old) had a low absorption, 3.3 and 1.8% respectively, corroborating their high need for additional elemental zinc (about 2 mg/kg/day). Three adult AE patients, all in their twenties, had a considerably lower need for extra zinc (about 0.2 mg/kg/day). Their zinc absorption ranged from 28 to 36% (mean 34%). In the controls the range was 27 - 65% (mean 43%). Turnover of retained 65Zn from day 8 - 30 was about 0.7% in the patient as well as in the control groups. Oral zinc therapy was withdrawn prior to the study. During the zinc-free period (3-7) a marked decrease in serum zinc and serum alkaline phosphatase values was noted in the two children with AE and they showed clinical evidence of zinc deficiency (angular stomatitis, scaling around finger nails, and irritability). None of the adult patients showed such evidence of impending zinc deficiency. One complained of exacerbation of facial acne, and another of pain in her feet. All symptoms disappeared promptly when oral zinc therapy was resumed.


Direct iodination of specific residues in crystals of yeast formylatable methionine-accepting transfer ribonucleic acid. Crystals of yeast formylatable methionine-accepting transfer ribonucleic acid (tRNAfMet) were iodinated by using a modification of Commerford's procedure [Commerford, S.L. (1971) Biochemistry 10, 1993]. Chromatographic analysis of nuclease digestion products showed that radioactive iodine binds covalently to the 5 position of three nucleotide residues: U8, C73, and C74. These three iodine substitutions were assigned to three peaks in a difference Fourier synthesis comparing the iodinated derivative with native tRNAfMet. In this way the positions of U8, C73, and C74 were marked in the crystal structure of yeast tRNAfMet, providing guidepoints for the interpretation of a 4.5-A electron density map.


Affinity labeling of a reactive sulfhydryl residue at the peptidyl transferase P site in Drosophila ribosomes. An affinity label has been prepared that is specific for the P site of a eucaryotic peptidyl transferase, that of Drosophila melanogaster. It has the sequence C-A-C-C-A-(Ac[3H]Leu) with a mercury atom added at the C-5 position of all three cytosine residues (referred to as the mercurated fragment). This label is an analogue of the 3' terminus of N-acetylleucyl-tRNA. The mercurated fragment binds specifically to the P site of peptidyl transferase. It participates fully in peptide bond formation as judged by its ability to transfer N-acetylleucine to puromycin with at least the same efficiency as a nonmercurated fragment. Once bound to the P site, the mercurated fragment reacts covalently with a ribosomal protein(s). This affinity-labeling process can be effectively competed by nonmercurated fragment, which indicates a site-specific reaction. The covalent attachment of the affinity label to a ribosomal protein(s) occurs through the formation of a mercury-sulfur bond, as judged by its lability in the presence of thiol reducing agents. The major ribosomal protein labeled at the P site of D. melanogaster was found to be a small, basic protein. The electrophoretic behavior of this protein parallels that of major P site proteins found in Escherichia coli ribosomes and in other eucaryotes. These results suggest conservation of some of the overall properties of the P site proteins from these organisms.


Sequence of histone 2B of Drosophila melanogaster. The complete sequence of histone 2B of Drosophila has been determined by using an improved Beckman sequenator. Comparing these data with those previously published by other investigators on the histone 2B of calf [Iwai, K., Hayashi, H., & Ishikawa, K. (1972) J. Biochem. (Tokyo) 72, 357--367], trout [Koostra, A., & Bailey, G. S. (1978) Biochemistry 17, 2504--2510], and Patella (a limpet) [van Helden, P. D., Strickland, W. N., Brandt, W. F., & von Holt, C. (1979) Eur. J. Biochem. 93, 71--78], it is possible to assess the evolutionary stability of this protein. There is little conservation of sequence in the N-terminal portion of the molecule (residues 1--26 numbering according to calf H2B), while the remainder of the protein, which we designate the C-terminal portion, is highly conserved. In the region of 27--125 residues, there are 9 substitutions in the composite data among the 98 positions, 8 of them conservative. These data indicate that very different selective pressures operate on the two different portions of the H2B molecule, implying the existence of two well-defined regions. Studies on the structure of the nucleosome by others have suggested that the C-terminal portion of H2B is involved in histone-histone interactions while the N-terminal portion is a relatively free "tail" binding to DNA. The sequence data indicate that the function of the C-terminal region of H2B requires considerable sequence specificity while that of the N-terminal region does not.


Purification and enzymatic properties of lysyl hydroxylase from fetal porcine skin. Lysyl hydroxylase from fetal porcine skin is shown to bind in a highly specific manner to aminoethyl-Sepharose 4B. When coupled to ammonium sulfate fractionation and DEAE-cellulose chromatography, chromatography of lysyl hydroxylase preparations on aminoethyl-Sepharose 4B has yielded a highly purified (greater than 95%) preparation of lysyl hydroxylase. The enzyme consists of two subunits with molecular weights of 70 000 and 115 000. The overall recovery of activity was 2.5%, yielding approximately to 3.5 mg of purified enzyme from 900 g of fetal porcine skin. The enzyme is more active at 30 degrees C than at 37 degrees C and has a pH optimum near 8.0. Both catalase and bovine serum albumin are required by the enzyme for maximum activity. The sulfhydryl reagents p-(chloromercuri)-benzoate, N-ethylmaleimide, and iodoacetamide are potent inhibitors of the enzyme, whereas dithiothreitol appears to be an activator.


Ribosome binding site analysis of ovalbumin messenger ribonucleic acid. The region of the ovalbumin messenger ribonucleic acid (mRNAov) molecule bound to the 40S ribosomal subunit and its associated initiation factors in the wheat germ cell-free translation system were isolated and characterized. Two mRNAov fragments, 87 and 92 nucleotides in length, were protected from T1 ribonuclease digestion by binding of guanosine 5',beta,gamma-methylenetriphosphate and were shown by hybridization and fingerprint mapping to be derived from the 5' end of mRNAov. Both these mRNAov fragments were of sufficient length to contain both the cap structure and the AUG initiation codon. Four T1-resistant oligonucleotides, prepared by direct digestion of mRNAov with T1 ribonuclease were also found to bind to the wheat germ 40S ribosomal subunit. Nucleotide sequence analysis of these oligonucleotides revealed (1) that they were not a subset of the ribosome binding fragments described above, (2) that they were derived from within the mRNAov molecule (one from within the coding region and three from the noncoding region at the 3' end of the mRNAov molecule), and (3) that three of the four mRNAov nucleotides contained 3'-terminal AUG trinucleotides. These data suggested that features of the mRNAov molecule in addition to the nucleotide sequence might be important in specifying the correct ribosome binding site for the initiation of protein synthesis. The amount of mRNAov bound to the wheat germ 40S ribosomal subunit in a preinitiation complex was found to vary inversely with the potassium ion concentration. Lowering the potassium concentration to levels suboptimal for translation also resulted in the protection of larger fragments of the mRNAov molecule derived from the same 5'-end region as the ribosome binding fragments described above. The ability of the cap analogue 7-methylguanosine 5'-phosphate (m7G5'p) to reduce the amount of mRNAov bound to the wheat germ 40S ribosomal subunit was found to depend directly on thepotassium concentration. Interestingly, the effects of potassium on the amount of mRNAov bound in a preinitiation complex and the inhibition of this binding by m7G5'p could be observed by changing the potassium concentration after binding had occurred. These data suggested that the interaction between the wheat germ 40S ribosomal subunit and mRNAov was very sensitive to the ionic environment.


Nucleic acid hybridization of highly repeated DNA in extracts of single Drosophila. We have developed and characterized a method for the rapid detection and quantitation of specific DNAs in partially purified extracts of single Drosophila. While the method should be applicable to a number of repetitious DNA sequences, we have used the polypyrimidine DNA sequences (TCTCT)n to develop this technique. Using hydroxyapatite chromatography, we were able to measure the amount of nucleic acid hybrid formed and to obtain a thermal elution profile of the hybrid formed in extracts of single flies. Under a variety of conditions, purified DNA and DNA in partially purified extracts gave essentially identical results. The procedure can be used to detect the presence of rare sequences, or to measure the relative abundance of a prevalent DNA species. 40 different wild type strains of Drosophila melanogaster were examined using this technique and all contain similar amounts of the same polypyrimidine/polypurine sequence. From a small scale screening of different laboratory stocks of D. melanogaster, a variant was found which formed more DNA-DNA hybrid with labelled polypyrimidine tracts than did wild type. The additional hybrid was distinguished by a lower thermal stability than the hybrid formed in wild type.


Isolation and characterization of cDNA complementary to transient maternal poly(A)+ RNA from the Drosophila oocyte. cDNA complementary to total oocyte poly(A)+ RNA from Drosophila melanogaster was enriched for sequences complementary to transient maternal sequences; that is, those sequences which disappear from the oocyte during subsequent. A seven- to ten-fold enrichment factor was obtained, from 5.3% to about 50% of the total cDNA. Kinetic analysis of this enriched fraction indicates that the transient maternal sequences include 44 +/- 14 different sequences.


Studies on temperature adaptation in Tetrahymena. Positional distribution of fatty acids and species analysis of phosphatidylethanolamine from Tetrahymena pyriformis grown at different temperatures. Phosphatidylethanolamine of 15 degrees C-grown Tetrahymena pyriformis (NT-I) cells contains more polyunsaturated fatty acids than 39.5 degrees C-grown cells. This increase in unsaturation is due to an increase in linoleic (C18 : 2) and linolenic (C18 : 3) acids, and a decrease in myristic (C14 : 0), palmitic (C16 : 0), palmitoleic (C16 : 1) and heptadecanoic (C17 : 0) acids. Compared with 39.5 degrees C-grown cells, the proportion of palmitic acid (C16 : 0) decreased in the 1-position as does at the 2-position in 15 degrees C-grown cells. On the contrary, there is a significant increase in linoleic (C18 : 2 delta 9, 12) and gamma-linolenic (gamma-C18 : 3) acids in the 1- and 2-positions, respectively. Phosphatidylethanolamine has been subfractionated into seven different diglyceride species. In 15 degrees C cells, the amounts of fractions 2 (1-linolenoyl-2-linoleoyl) and 3 (1-linolenoyl-2-palmitoleoyl, 1-linolenoyl-2-oleoyl) increased while there was a great decrease in subfraction 7 (1-myristoyl-2-palmitoleoyl, 1-palmitoyl-2-palmitoleoyl). Since subfractions 1 and 2 contain over 70% linoleic (C18 : 2) and linolenic (C18 : 3) acids, these fractions might be composed mainly of 1-linolenoyl-2-linolenoyl and 1-linolenoyl-2-linoleoyl molecular species at 15 degrees C. These data support evidence that phosphatidylethanolamine would play a principal role as an acceptor of acyl chains for temperature acclimation.


Studies on the relationship between glycerophosphoglycolipids and lipoteichoic acids. IV. Trigalactosylglycerophospho-acylkojibiosyldiacylglycerol and related compounds from Streptococcus lactis Kiel 42172. Streptococcus lactis Kiel 42172 contains at least six unusually polar glycerophosphoglycolipids. The predominant one was composed of D-galactose, D-glucose, glycerol, acyl groups and phosphorus in a molar ratio of approx. 3 : 2 : 2 : 3 : 1. By analysis of the breakdown products of HF hydrolysis and Smith-degradation the structure was established to be [Galp (alpha 1 leads to 6)Galp(alpha 1 leads to 3)-sn-glycero(2 comes from 1 alpha Galp)-1-phospho] leads to 6Glcp(alpha 1 leads to 2), acyl leads to Glcp(alpha 1 leads to 3)-acyl2Gro. By HF hydrolysis the other compounds were shown to be in the main also derivatives of GroP leads to 6Glc(alpha 1 leads to 2), acyl leads to 6Glc(alpha 1 leads to 3)acyl2Gro but they released as water-soluble glycosides Gal(alpha 1 leads to 2)Gro, Gal(alpha 1 leads to 3)Gro, Gal(alpha 1 leads to 3)Gro(2 comes from 1 alpha Gal), Gal(alpha 1 leads to 6)Gal(alpha 1 leads to 3)Gro and Gal(alpha 1 leads to 6)Gal-(alpha 1 leads to 6)Gal(alpha 1 leads to 3)Gro(2 comes from 1 alpha Gal), respectively. In the lipid extract Glc(alpha 1 leads to 2), acyl leads to 6Glc(alpha 1 leads to 3)acyl2Gro and GroP leads to 6Glc(alpha 1 leads to 2), acyl leads to 6Glc(alpha 1 leads to 3) acyl2Gro were also observed. This set of compounds is proposed to constitute a biosynthetic series reflecting the individual steps in the synthesis of the lipoteichoic acid of Streptococcus lactis Kiel 42172 which is made up by the same lipid anchor and a non-classical poly(galabiosyl, galactosyl glycerophosphate)-chain (Koch, H.U. and Fischer, W. (1978) Biochemistry 17, 5275--5281).


Enzymatic sulfation of galactosyl- and lactosylceramides in cell lines derived from renal tubules. 1. The renal cell lines, JTC-12 and MDCK, not only synthesize galactosylceramide 3-sulfate and lactosylceramide 3'-sulfate in vivo, but also contain enzymes that catalyze the transfer of sulfate to galactosylceramide and lactosylceramide in vitro. 2. Concentration of cations necessary for maximum sulfotransferase activity occurred at 40 mM Ca2+ with galactosylceramide and 15 mM Ca2+ with lactosylceramide as the substrate. Na+ was also found to stimulate the sulfation of galactosylceramide, but was slightly inhibitory for the sulfation of lactosylceramide. 3. The products of the in vitro assay mixture were characterized as galactosylceramide 3-sulfate and lactosylceramide 3'-sulfate by a variety of TLC separations. 4. The apparent Km of JTC-12 cells for galactosylceramide was 17 microM, while that for lactosylceramide was 82 microM. The Km values of MDCK cells were comparable to those of JTC-12 cells. Competition studies suggested that galactosylceramide and lactosylceramide were sulfated by a single enzyme in both cell lines.


Co-oxidation of carotenes requires one soybean lipoxygenase isoenzyme. The type II lipoxygenase (optimum pH 6.5) from soybeans was purified and separated into two fractions either by chromatography on DEAE-Sephadex or by isoelectric focusing. In the presence of linoleic acid and oxygen both fractions co-oxidise canthaxanthine or beta-carotene as effectively as a combination of these fractions. Oxygenation of linoleic acid and co-oxidation of canthaxanthine by type II lipoxygenase is stimulated by 13-hydroperoxy-cis-9,trans-11-octadecadienoic acid but not by 13-hydroxy-cis-9,trans-11-octadecadienoic acid or 9-hydroperoxy-trans-10,cis-12-octadecadienoic acid.


The effect of temperature on unsaturated fatty acid loss in Tetrahymena pyriformis. Cultures of Tetrahymena pyriformis W incorporate exogenous 3-[14C]-cilienic acid and gamma-[1(-14)C] linolenic acid, terminal products of unsaturated fatty acid synthesis, into glycerophosphatides without randomization of the radiolabel. There was no difference in the rate of loss of each of the two acids at 15 or 28.5 degrees C. Differential turnover of these fatty acids, therefore, does not appear to be the cause of the shift in fatty acid pattern observed with temperature reduction.


9-LR-linoleyl hydroperoxide, a novel product from the oxygenation of linoleic acid by type-2 lipoxygenases from soybeans and peas. Type-2 lipoxygenases from soybeans and peas, which have a pH optimum of 6--7 were examined for oxygenation activity at pH 9.0. The reaction velocity was found to be strongly dependent on substrate concentration. At higher substrate concentrations an inhibitory effect was observed, which is connected with the occurrence of a kinetic lag phase. On incubation of linoleic acid at pH 9.0 with either of these enzymes predominantly 9-LR-hydroperoxy-10-trans,12-cis-octadecadienoic acid is formed. The similarity of the product specificity with that of prostaglandin synthetase is discussed in view of the formation of prostaglandin-like substances by soybean lipoxygenase-2 (Bild, G.S., Bhat, S.G., Ramadoss, C.S. and Axelrod, B. (1978) J. Biol. Chem, 253, 21--23).


Proteolytic fragmentation of myosin: location of SH-1 and SH-2 thiols. The heavy chain fragmentation pattern of native myosin when digested by proteolytic enzymes is influenced by such conditions as the nature of the proteolytic agent, ionic strength and presence or absence of divalent cations. HMM and S-1 produced by digestion of 14CNEM-labelled myosin under various conditions were analyzed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis. Purified samples of these species were digested under controlled conditions by chymotrypsin and trypsin and a comparison of the observed heavy chain fragmentation patterns led to a sequential arrangement of the proteolytic fragments. The main features of this arrangement are the following: a 21K molecular weight tryptic peptide is found at the N-terminal side of myosin heavy chain. Adjacent to it is a 48K peptide, then a 19.5K peptide containing the two SH-1 and SH-2 thiols. These three peptides constitute the heavy chain of S-1. Adjacent to this S-1 heavy chain is a tryptic (and also chymotryptic) 40K peptide. The rest of the HMM heavy chain on the C-terminus is a sequence susceptible to both chymotrypsin and trypsin attack yielding an undefined number of small peptides.


Evidence for the resistance of thyrotropin-releasing hormone (TRH) and pseudo-hormone, pyroglutamyl histidyl-amphetamine, to degradation by enzymes of the digestive tract in vitro. TRH and pseudo-hormone (pyro Glu-His-amphetamine) were submitted to the digestion of chymotrypsin and prolidase and independently to the digestion of enzymes of the digestive track: pepsin (stomach), pancreatins (pancreas) and enzymes extracted from the intestinal mucosa (small intestine). Using thin layer chromatography and high voltage electrophoresis techniques to detect enzymic digestion products, only intact TRH and pseudo-hormone were found, indicating that both entities were, under the conditions used, resistant to in vitro digestion by enzymes of the digestive tract.


Hemolysis with red cell covered surfaces. Polypropylene (PP) disks activated by exposure to ammonia glow discharge were used as substrates for red cells deposited from saline suspensions. In some cases these cell-coated disks were further treated by glutaraldehyde to bind the cells more strongly. Each disk was used in a rotational blood-shearing device to induce hemolysis, which was compared with that induced by virgin PP. When the cell coating was uniform and dense from a single settling of cells, the glutaraldehyde-fixed surfaces were about 10% less hemolytic than PP. Non-fixed cells detached from outer regions of the disk and apparently contributed additional hemolysis in the process. Secondary layers of settled cells, both fixed and non-fixed, also proved to be more hemolytic than PP. Coatings of gamma-globulin reduced hemolysis relative to uncoated cell surfaces. The best performance was that of activated surfaces without cells, with hemolysis about 20% less than PP. Tests of cells hemolyzing during detachment in a saline medium suggested that hemolysis of whole blood involves a series of brief attachment/detachment events at the solid surface.


Bonding erythrocytes to plastic substrates by glow-discharge activation. A procedure was developed for attaching erythrocytes to various plastics. Plastic disks were exposed to ammonia glow discharge (AGD) and found thereafter to form weak bonds with red cells settling from a dilute saline suspension poured over the disks. When the disks were used to produce a rotating shear flow, complete cell detachment was observed for stresses exceeding 34 dynes/cm2. Far stronger bonding was achieved by treating AGD-bonded cells with glutaraldehyde, and subsequent shearing failed to detach any cells (maximum stress 58 dynes/cm2). The proposed chemical mechanism involves attachment of -NH2 groups on the plastic during AGD, and formation also of carbonyl oxygen later, to provide sites for hydrogen bonding with the cell membrane. Subsequent glutaraldehyde exposure produces crosslinking between disk and cell, as well as fixing the cell in the normal way. Thus, the initial step succeeds in attaching erythrocytes in a deformable condition but rather weakly, while the second step produces rigid cells which are also bonded more strongly.


Convenient method for studying enzyme kinetics. A convenient method for enzyme kinetic studies is introduced. The method includes identification of reaction mechanism and estimation of the associated kinetic constants with a minimum number of experiments. The application of the method is illustrated by using literature data. Factors limiting the application of this method are also discussed.


[Transforming activity and the oncogenicity of simian adenovirus SA7 DNA]. The transforming and oncogenous activity of uncleaved DNA of simian adenovirus SA7 (AdSA7) and the products of its restriction by endonucleases R. Bam HI and R. SalI was studied. It was shown that uncleaved virus DNA transformed the rat kidney cells and rat embryo fibroblasts and induced tumors in newborn hamsters. AdSA7 DNA, hydrolysed by R. Bam HI, posessed the transforming activity. The mixture of DNA fragments, obtained after hydrolysis by R. SalI was oncogenous in hamsters.


[Immunochemical study of heterogenous antigens Y. pestis EV similar to human red cells]. Franctions containing heterogenous antigens Y. pestis EV similar to human red cells can be obtained by the method of immunosorption of antigens by fixed antibodies on polyacrylamide gel.


[Immunoglobulin receptors of group A Streptococcus]. It was shown in the gel precipitation tests that absorption of human and rabbit IgG or Fc-fragments obtained from human IgG group A streptococcal cultures results in inhibition of the reactions of these preparations with immunoglobulin sera. The reactions of F(ab')2-fragments with the corresponding sera are not inhibited during their absorption by the same cultures. The results obtained support the presence in a number of group A streptococcal cultures of immunoglobulin receptors (Ig-receptors) capable of reacting with Fc-parts of human and rabbit IgG. Pepsin treatment destroys Ig-receptors. These receptors could not be found by the method used in hydrochloric acid extracts prepared from streptococci containing the receptors. The method can be applied for determination of Ig-receptors in streptococcal cultures.


[Population of L-forms of Bacillus subtilis studied in Ficoll density gradients]. The distribution of cells in the population of L-forms of Bas. subtilis was analysed by isopicnic centrifugation in density gradient of ficoll. Two main fractions of L-forms different in their density were found. The study of the fractions by various methods indicated that a considerable part of the L-forms population is presented by unviable cells of diverse size with fragments of genome or without DNA.


[Counter-flow isotachophoresis on cellulose acetate membranes. Role of electroendosmosis]. A variant of counter-flow isotachophoresis of proteins on cellulose acetate membranes is proposed. The liquid counter-flow is created by electroendosmosis in the membrane. Proteins are concentrated at the Kolrausch boundary during isotachophoresis in the presence of ampholytes. The method permits one to make microanalysis of proteinic mixtures in diluted solutions, and it can be used in combination with immunodiffusion and immunoelectrophoretic methods of antigenic protein detection.


[Estimation of the contribution of thiophosphamide-induced chromosomal aberrations to perimplantation embryonic mortality in mice]. Thiotepa-induced perimplantation mortality in mice was analysed cytologically. The contribution of induced chromosomal aberrations to preimplantation embryonic death was studied. Under the influence of 1.25 mg/kg of thiotepa on late spermatids the stop of cleavage division was found at 2--16 cell stage in 27% of the cases and at 17--22 cell stage in 11.9% (totally in 38.9%) as compared to 6% in the controls. The cytogenetic analysis of embryos consisting of 2--16 cells revealed gross structural chromosomal aberrations in 75% metaphases suitable for analysis.


The effects of sodium cromoglycate on lung irritant receptors and left ventricular cardiac receptors in the anaesthetized dog. 1 The time from the injection of sodium cromoglycate 10 to 50 mug/kg into a saphenous vein, the cervical carotid arteries, the left ventricle and the aortic arch, to the onset of reflex hypotension has been measured in anaesthetized dogs. The shortest latency was 16.9 s on injection of sodium cromoglycate into the left ventricle.2 Instillation of 2% lignocaine into the pericardium of an anaesthetized dog blocked the reflex hypotensive response to sodium cromoglycate (10 to 50 mug/kg i.v.), and also prevented sodium cromoglycate (100 mug/kg) from reversing reflex bronchoconstriction induced by inhalation of an aerosol of histamine.3 The effect of sodium cromoglycate (100 mug/kg i.v.) on resting discharge and histamine-induced discharge (20 mug/kg i.v.) of five lung irritant receptors in five anaesthetized dogs has been studied. Sodium cromoglycate (100 mug/kg i.v.) did not affect the resting discharge of these receptors or their ability to respond to histamine.4 Sodium cromoglycate (100 mug/kg i.v.) increased the rate of discharge of three receptors found in the endocardium of the left ventricle of the canine heart. A solution of sodium cromoglycate (0.1%) was applied topically to one receptor and its rate of discharge was increased.5 It is suggested that in the dog, sodium cromoglycate produces reflex hypotension and reverses histamine-induced reflex bronchoconstriction by activating receptors in the left ventricle of the heart.


Assessment procedures used in studies on long-stay patients: a survey of papers published in the British Journal of Psychiatry. Every article published in the British Journal of Psychiatry between 1945 and 1974 was reviewed that provided original information derived from the direct assessment of long-stay patients. The 225 articles were examined for details of assessment practice and characteristics of the patients studied. Most studies used a very restricted range of assessment methods, with rating scales, used most frequently. Many used wholly unstandardized methods, and the description of patient characteristics was inadequate. A number of improvements in assessment practice are suggested.


Ossification of the posterior longitudinal ligament of the spine. "The Japanese disease" or ossification of the posterior longitudinal ligament of the cervical spine was initially thought to affect only the Japanese people. Non-Japanese Asian and Caucasian cases have been reported but appear to be exceedingly rare. The reason is unknown. Although the disease appears to be generally benign, it can cause a myelopathy. We present here 16 non-Japanese patients in Singapore who were found to have ossification of the posterior longitudinal ligament in the cervical, thoracic and lumbar spine.


Acute pulmonary distress following high-dose irradiation of the upper half of the body. Two cases of acute pulmonary distress following very shortly after irradiation of the upper half of the body are described. The first occurred one hour and forty minutes after irradiation and led to the patient's death three hours after irradiation. The second occurred 15 minutes after irradiation and was successfully treated with hydrocortisone and Frusemide. The features are the sudden onset of dyspnoea and cyanosis associated with pyrexia. Auscultatory evidence of pulmonary oedema was not apparent initially. The response to steroid alone is transitory. High single-dose pulmonary irradiation causes transudation of extracellular fluid and diapedesis of red cells from the alveolar capillaries. The supervention of these effects on severly compromised pulmonary function can well prove dangerous.


Contrast enhancement of high-energy radiotherapy films. An order-of-magnitude improvment in the contrast of high-energy localization and verification films has been achieved through the application of a simple, inexpensive, contrast enhancement technique. The method involves making reversal contact "prints" of the original film onto ordinary X-ray fi-m with equipment commonly available in any radiotherapy department. This results in "gamma multiplication". The theory as well as several applications of this effect are presented.


O-diphenoloxidase concentrations in leprosy. O-diphenoloxidase activity was studied in 15 patients with lepromatous leprosy, 15 with tuberculois leprosy, and 15 controls. O-diphenoloxidase isolated from skin and serum samples of patients with lepromatous leprosy had the specificity of a bacterially derived enzyme and not that of a mammalian-derived enzyme. Only the patients who had lepromatous leprosy for over two years showed enzyme activity in serum, though all showed it in skin tissue. O-diphenoloxidase activity in serum may be a useful diagnostic marker of lepromatous leprosy.


Reduction in polypharmacy for epilepsy.

Reduction in polypharmacy for epilepsy. A two-year prospective study of 40 adult outpatients with chronic epilepsy was carried out in which blood drug concentrations were monitored, and anticonvulsant polypharmacy was reduced to treatment with a single drug in 29 patients (72%). In the year after the reduction of treatment the control of seizures was improved in 16 patients (55%), unchanged in eight(28%), and worse in five (17%). Mental function was improved in 16 (55%). The main reason for failure to reduce to or maintain treatment with a single drug was exacerbation of seizures during the difficult withdrawal period, especially in patients with frequent seizures, taking several drugs, or with additional neuropsychological handicaps. It is more difficult to reduce polypharmacy than to avoid it in the first place. Polypharmacy may sometimes aggravate control of seizures.


Bronchodilator effect of sodium cromoglycate and its clinical implications. The bronchodilator effect of sodium cromoglycate (SCG) solution was investigated. Twenty asthmatic children aged 6-15 years (mean 11.3) were examined and the action of SCG compared with that of salbutamol and placebo (water). SCG produced a significantly raised peak expiratory flow rate (PEFR) before exercise, which reached a maximum immediately after exercise. The bronchodilatation was sustained for up to four hours, when the PEFR was still significantly above the resting value. This effect was comparable in degree and duration with that of salbutamol. In contrast, placebo produced insignificant bronchodilatation before exercise but significant albeit short-lived bronchodilatation after exercise, which is the characteristic response of the asthmatic to a short period of exercise. This powerful bronchodilator action of SCG and its equally potent inhibitory action on exercise-induced bronchoconstriction was achieved by administering the solution via an efficient nebuliser. In order to achieve maximum clinical effect the SCG must, therefore, be given in this form.


Role of residual insulin secretion in protecting against ketoacidosis in insulin-dependent diabetes. The role of preserved beta-cell function in preventing ketoacidosis in type I insulin-dependent diabetes was assessed in eight patients with and seven patients without residual beta-cell function as determined from C-peptide concentrations. After 12 hours of insulin fatty-acid, and glycerol concentrations were all significantly higher in patients without beta-cell function than in those with residual secretion. Mean blood glucose concentrations reached 17.2 +/- SE of mean 1.3 mmol/l (310 +/- 23 mg/100 ml) in the first group compared with 8.8 +/- 1.4 mmol/l (159 +/- 25 mg/100 ml) in the second (P less than 0.01), while 3-hydroxybutyrate concentrations rose to 5.5 +/- mmol/l (57 +/- 5 mg/100 ml) and 1.4 +/- 0.3 mmol/l (15 +/- 3 mg/100 ml) in the two groups respectively (P less than 0.01). Individual mean C-peptide concentrations showed a significant inverse correlation with the final blood glucose values (r = -0.91; P less than 0.02). These findings strongly suggest that even minimal residual insulin secretion is important for metabolic wellbeing in diabetes and may prevent the development of severe ketoacidosis when insulin delivery is inadequate.


Significance of early increase in stable and radioactive plasma calcium after parathyroidectomy in primary hyperparathyroidism. Early effects of parathyroid hormone (PTH) deficiency were studied in 12 patients with primary hyperparathyroidism due to single parathyroid adenoma by following the precise time course of changes in plasma calcium (Ca) and immunoreactive parathyroid hormone (IPTH) after parathyroid surgery and by prelabeling 2 patients with radiocalcium (Ca*). Surgical removal of the adenoma was immediately followed by a sudden increase in plasma Ca which preceded the usual fall. The increase in plasma Ca commenced simultaneously with the fall in iPTH and was accompanied by a parallel increase in specific activity (sp. act.) of plasma Ca*. Specific activity continued to rise for 2 h in both prelabeled patients, whereas blood calcium was already falling thereafter reaching a markedly low removal rate constant as long as plasma Ca decreased. When plasma Ca began to rise, sp. act. resumed a descending course. Our findings indicate that the initial hypercalemia depends on PTH withdrawal and results from a rapid flux into general extracellular fluid (ECF) of calcium coming from a compartment with higher sp. act., contained within the miscible pool, immediately followed by a reduction in calcium transfer from bone. These results suggest that acute PTH deficiency determines an outflow of calcium from bone cells and support the theory that PTH initiates its action by modifying their intracellular calcium content.


Comparison of the effects of 1,25-dihydroxycholecalciferol, prostaglandin E2, and osteoclast-activating factor with parathyroid hormone on the ultrastructure of osteoclasts in cultured long bones of fetal rats. The effects of 1,25-dihydroxy vitamin D3 [1,25(OH)2D3], prostaglandin (PGE2), and osteoclast-activating factor (OAF) on the size of osteoclasts, nuclei, ruffled borders, and clear zones in cultured long bones of fetal rats were quantitated. In addition, the number of osteoclasts in the bones was counted and the release of calcium from the bone into the culture medium was determined. These data were compared with the corresponding effects of parathyroid hormone (PTH). All agents tested increased the size of the ruffled borders significantly after 3 h, the size of the clear zones after 12 h, and the size of the cells after 12-24 h. No important differences in sizes were noticed between the agents tested or between the agents and PTH. The number of osteoclasts was increased after 24 h of treatment with PTH, but not after the other agents. Calcium release was significantly increased for all agents between 12 and 24 h. It is concluded that bone resorption by 1,25(OH)2D3, OAF, and PGE2 is mediated primarily by increased activity of existing osteoclasts similar to PTH activation.


A cat model for the evaluation of mechanisms of bone resorption: induction of bone loss by simulated immune complexes and inhibition by indomethacin. When simulated immune complexes (SIC) (heat-aggregated IgG) possessing many of the properties of true antigen-antibody complexes were injected via the root canal into the periapical tissues of cat maxillary cuspids, radiographically and histologically evident bone resorption occurred at these sites within 7 days. Bone loss was accompanied in all cases by inflammation of the surrounding collagenous connective tissues and was characterized by the presence of osteoclasts. Bone resorption, but not the accumulation of inflammatory cells, was blocked by the systemic administration of indomethacin, an inhibitor of prostaglandin synthetase. The most likely explanation is that SIC-activated mechanisms such as the complement cascade, prostaglandin synthesis, and neutrophil degranulation were responsible for the bone loss. The minor inflammation and bone loss that followed the repeated injections of BSA and of monomeric IgG can best be explained as a response to trauma. The data presented establish that the cat maxillary cuspid is a useful model in which to explore the mechanism underlying pathological bone resorption.


Comparison of the effects of a potent synthetic analog of bovine parathyroid hormone with native bPTH-(1-84) and synthetic bPTH-(1-34) on bone resorption and collagen synthesis. An analog of bobine PTH [nle-8, nle-18, tyr-34 bPTH-(1-34) amide, (PTH-Ana)] which is a potent stimulator of renal adenylate cyclase has been compared with the native hormone bPTH-(1-84) and the biologically active amino terminal portion, bPTH-(1-34), for its effects on bone resorption and bone collagen synthesis in organ culture. All three compounds stimulated the release of previously incorporated 45Ca from cultured fetal rat long bone shafts with similar dose-response curves at 10(-9) to 3 X 10(-8) M. All three compounds inhibited bone collagen synthesis as measured by incorporation of proline into collagenase digestible protein, whereas incorporation into noncollagen protein was not inhibited. The effects were dose related and decreases in percent collagen synthesis were significant at 10(-9) M. Thus PTH-Ana appears to have the same effects on bone resorption and collagen synthesis as bPTH-(1-84) and (1-34) and is likely to be a valid probe for investigating PTH receptors in bone as well as in kidney.


Early osteogenesis in compact bone isografts: a quantitative study of contributions of the different graft cells. Isografts of cortical bone were tranplanted subcutaneously in the rat and the rate of osteogenesis 12 to 14 days later was assessed by measurement of 85Sr uptake and by histology. Some grafts were implanted complete whereas others had had one or more of their cellular components (viz. periosteum, endosteum, osteocytes, marrow) removed by mechanical or enzymatic pretreatment. From an analysis of the differences in osteogenesis between grafts devoid of different combinations of cellular components, the contribution of each component to osteogenesis was determined. The results indicate that the endosteal lining cells and marrow stroma together produce more than half of the new bone, the periosteal cells contribute about 30%, the osteocytes possibly make a small (10%) contribution, and the free, hemopoietic cells of the marrow make no significant contribution. Evidence about the relative contributions to osteogenesis of graft and host cells is reviewed and the possible osteogenetic role of bone marrow is discussed.


Ectopic calcification in lambs from feeding the plant Cestrum diurnum. Hypercalcemia and ectopic calcification were induced in 5 lambs by supplementing the diet with the dried leaves of the plant Cestrum diurnum, for 8 to 9 weeks. Lambs developed mineralization of blood vessels, heart, kidneys, and lungs. These tissues were examined by light and electron microscopy. In the vascular tissue there was calcification of elastic fibers in the hyperplastic intima and the media, along with mineralization of mitochondria of aortic smooth muscle cells. Myocardial cells and their mitochondria were mineralized. In the kidney, there was calcification of the epithelium of the distal convoluted tubules and collecting tubules, Bowman's capsule, and the mesangial cells of the glomeruli. In the lung, there was mineralization of the alveolar septal walls and the bronchi and bronchioles. Feeding of the calcinogenic plant to lambs caused extensive soft tissue calcification. Results of the study indicated that degeneration was the early soft tissue lesion in this plant toxicity.


Cathepsin D: ultra-immunohistochemical localization in dentinogenesis. Cathepsin D was purified from rat liver using a new affinity chromatographic method, based on the coupling to the specific inhibitor pepstatin. This preparation was used for the production of specific antibodies from rabbit. The purified IgG fraction was conjugated to horseradish peroxidase in a two-step coupling procedure and used for electron microscopic immunohistochemistry of the odontoblast-predentine region of the rat incisor. Precipitates, indicating the presence of cathepsin D, were seen in the odontoblast, odontoblast process, and in the extracellular unmineralized matrix, the predentine. The observations are discussed in relation to proteoglycan degradation at the mineralization front simultaneous with crystal formation, and in relation to the function of lysosomal enzymes in the turnover of connective tissue.


Intracellular divalent cations and neuronal excitability. Itracellular injections of Mg into cat spinal motoneurones have a depolarizing action, associated with a fall in input conductance, and depression of the postspike hyperpolarizing after-potential (a.h.p.) as well as its underlying conductance increase. There is also an increase in excitability, sometimes leading to outright discharge, and a change in the current-firing relation: the normal primary range is largely abolished and the firing appears to have the characteristics of the normal secondary range. Intracellular effects of Mg are thus mainly opposite to those of Ca, possibly owing to competition at sites where Ca activates K channels. Intracellular injections of Mn also tend to depress the a.h.p. but have relatively little effect on resting potential and conductance, or action potentials. Co also depresses the a.h.p. but has a more pronounced depolarizing action, and produces particularly strong depression of action potentials. By contrast intracellular Sr tends to raise the membrane conductance and has a mild hyperpolarizing effect. During the injection of Sr, a.h.p's are depressed but this is followed by a rebound of increased a.h.p. amplitude and conductance. Unlike the other divalent cations tested, Sr strongly depressed excitatory postsynaptic potentials. In most respects Sr appears to behave like Ca.


Immunohistochemical localization of thyrotropin-releasing hormone (TRH) in skin of Rana pipiens. Using immunofluorescent techniques thyrotropin releasing hormone (TRH) is demonstrated in skin of Rana pipiens and R. catesbeiana. The immunofluorescent-TRH is localized in all cell layers of the epidermis and in the epithelium lining the various cutaneous glands, but not in the dermal layer.


Current concepts in the immediate management of acute spinal cord injuries. The management of acute spinal cord injuries has changed considerably during the past 10 years owing to new information about the pathophysiology of cord trauma and new diagnostic and treatment methods. It is now known that the cord suffers not only from the immediate physical effects of trauma, but also from secondary pathologic processes, such as ischemia and edema, which are treatable in the first few hours after injury. New neuroradiologic and neurophysiological techniques, such as the recording of the somatosensory evoked potential, increase the accuracy of diagnosis and prognosis in the acute phase. Current immediate treatment includes the administration of steroids and mannitol, with careful attention to respiratory and cardiovascular homeostasis, to overcome post-traumatic ischemia and edema, and immobilization of the spine with devices such as the halo. New surgical procedures are used in selected cases to improve neurologic recovery, to provide rigid immobilization of the spine or to allow earlier mobilization of the patient. The care of spinal cord injuries in the acute phase is facilitated by multidisciplinary units.


In vitro induced pinocytotic activity by a juvenile hormone analogue in oocytes of Drosophila melanogaster. Pinocytotic activity has been analyzed in Drosophila oocytes following either in vivo or in vitro exposure to horseradish peroxidase. The enzyme tracer gains access to the yolk spheres only when supplied to the oocyte in vivo. In oocytes cultured in vitro, peroxidase remains restricted to the residual coated vesicles and to the tubular profiles formed in excess in the cortical ooplasm. In an attempt to induce peroxidase uptake by oocytes cultured in vitro, various incubations were tested. Among these, hemolymph from both sexes is capable of promoting peroxidase uptake up to a level comparable to that detectable in vivo. On the other hand, fat body extracts fail to promote such cellular activity. Finally, the juvenile hormone analogue ZR-515 is shown to be the only factor required to promote pinocytotic activity under the experimental conditions tested. The observations are interpreted to indicate that vitellogenin has no inductive role on pinocytosis but simply acts by adhering to the forming coated vesicles which in turn are produced by the oolemma in response to the action of juvenile hormone.


Water compartments in living, glycerinated and fixed skeletal muscles of the frog. The kinetics of water replacement with heavy water (deuterium oxide) in the gastrocnemius and sartorius muscles of the frog under isotonic conditions, studied both gravimetrically and by infrared photometry, reveals three water compartments: (i) non-exchangeable (approximately 80 ml/kg fresh weight), (ii) slowly exchanging (approximately 500 ml/kg fresh weight), (iii) rapid exchanging--extracellular (approximately 200 ml/kg fresh weight). Exposure to both glycerol and glutaraldehyde increases the permeability coefficients and the amount of rapid exchanging water; glutaraldehyde also increases the amount of non-exchangeable water. Approximately 90% of the water is kept in the tissue only by weak intermolecular forces, the energies of which amount to 1 kcal/mol. The amount of non-exchangeable water is equivalent to about six continuous adsorption layers covering the myofilaments. Approximately 70% of the tissue water appears to be replaced by glutaraldehyde during standard fixation.


Alpha-amanitin-resistant D. melanogaster with an altered RNA polymerase II. Following EMS mutagenesis we recovered a mutant of D. melanogaster that grows at concentrations of alpha-amanitin lethal to wild-type. To our knowledge this mutant represents the first example of an amanitin-resistant eucaryotic organism. The amanitin resistance of the mutant (AmaC4) is due to an alteration in its DNA-dependent RNA polymerase II, which is approximately 250 times less sensitive to inhibition by amanitin than the wild-type polymerase II whether tested in nuclei, in partially-fractionated extracts or as a highly purified enzyme. While the wild-type enzyme activity is inhibited 50% by 2.1 x 10(-8) M alpha-amanitin, inhibition of 50% of the AmaC4 RNA polymerase II activity requires a toxin concentration of 5.6 x 10(-6) M. The mutation responsible for the amanitin resistance of AmaC4 is on the X chromosome near the vermillion locus.


Precursor-specific nucleotide sequences can govern RNA folding. An immediate precursor of 5S ribosomal RNA (rRNA) from Bacillus subtilis has 21 and 42 nucleotide precursor-specific segments associated with its 5' and 3' termini, respectively. On the basis of its nucleotide sequence, predicted secondary structure and location in the rRNA transcriptional unit, the 3' precursor element apparently functions during the termination of transcription. A portion of the 5' precursor element is shown to facilitate the native folding of the mature domain of the precursor. Precursor 5S rRNA molecules which lack the 5' terminal 8-9 nucleotides of the 5' precursor elements were fabricated. These abbreviated constructs assume a non-native conformation, as revealed by their behavior during polyacrylamide gel electrophoresis. The aberrant conformation is evidently forced upon the abbreviated constructs by the residual 5' precursor sequence, since its removal by the maturation endonuclease RNAase M5 precipitates the reordering of the mature domain into its native conformation. Inspection of the nucleotide sequence of the 5S precursor suggested the nature of the conformational aberration, and gel electrophoresis analyses of limited nuclease digests of end-labeled precursors in the native and aberrant conformations are consistent with the derived model. We conclude taht the 5' terminal six nucleotides in the intact 5S precursor assist in the folding of the mature domain by forming a base-paired duplex with neighboring nucleotides, thereby preventing that adjacent sequence from engendering the abnormal conformation. The involvement of precursor-specific sequences and conformational dynamics in RNA function are discussed.


Expression of ribosomal DNA insertions in Drosophila melanogaster. Approximately half of the ribosomal genes on the X chromosome of Drosophila melanogaster are interrupted by an insertion of type 1. Nuclear RNA from D. melanogaster embryos was transferred to DBM paper and hybridized with cloned type 1 insertion sequences. With a DNA fragment derived specifically from large insertions, transcripts were detected between 5 and 10 kb. These insertion transcripts represent less than one RNA molecule per nucleus, which is more than three orders of magnitude below the concentration of nascent rRNA chains, as determined by kinetics of hybridization. With a DNA fragment derived from the right end of large insertions which is also complementary to short insertions, more discrete RNA bands appeared with sizes between 1 and 8.5 kb, representing altogether about 13 RNA molecules per nucleus. Insertion transcripts large enough to be potential precursors to 28S rRNA represent less than one molecule per nucleus. It was shown by sandwich hybridization that at least some of the insertion transcripts are derived from rDNA. No significant difference was found between insertion transcripts in RNA extracted from ovaries, embryos, larvae, pupae or adult flies. Unless a mechanism other than splicing is involved, ribosomal genes with insertions cannot contribute significantly to the synthesis of 28S rRNA. A cytoplasmic RNA approximately 1 kb long, which is complementary to a short insertion and to ribosomal gene sequences flanking both sides of the insertion, was found. The abundance of this short unspliced RNA is about 50 molecules per embryo cell.


The clustered and scrambled arrangement of moderately repetitive elements in Drosophila DNA. An examination of cloned Drosophila DNA has revealed large clusters of densely spaced, short (less than or equal to 1 kb), moderately repetitive elements. Different clusters have many of the same repetitive elements, but these elements are arranged differently in each cluster. It is improbable that this clustered arrangement can be detected by conventional reassociation kinetic and electron microscopic techniques, but it can be detected and features of its fine structure can be determined by a two-dimensional version of Southern's blotting technique. The genomic organization of these clustered repetitive elements was investigated by hybridizing restriction fragments of cloned DNA to polytene chromosomes, to filter-bound recombinant DNA clones and to Southern blots of total Drosophila DNA. These studies demonstrated that clusters occur in euchromatic regions of the chromosomes and that at least one of the clusters has the same repetitive element organization in cloned and in chromosomal DNA. These studies also demonstrated that copies of the elements from one cluster are scattered in at least 1000 chromosomal regions. These regions appear to have differing concentrations of repetitive DNA, but together they account for a large fraction of Drosophila's moderately repetitive DNA. Aside from indicating the genomic organization of cluster elements, this work has identified cluster elements throughout a 9 kb region neighboring one of the heat shock genes, throughout the intron of the major rDNA repeat and within the apparently transposable element, 412.


Genetic and molecular analysis of the 87A7 and 87C1 heat-inducible loci of D. melanogaster. Two different types of heat-inducible sequences are found at the cytogenetic loci 87A7 and 87C1 of D. melanogaster. One of these codes for the 70,000 dalton heat shock protein (hsp 70) and is found at both loci. The other type of sequence (alpha beta) codes for an RNA of unknown function and is found only at 87C1. We have completed a study of the organization of the two loci, using deficiencies that delete one or other locus, and have estimated the number of the hsp 70 genes at each locus. Thus in at least three strains of files there are a total of five coding sequences, three at 87C1 and two at 87A7. Restriction mapping of the coding regions at the two loci reveals that each of the two cytogenetic loci has its own characteristic coding sequence. The overall organization of the two loci appears to differ considerably. The alpha beta and hsp 70 heat-induced sequences at 87C1 are closely linked and are contained within two Eco RI restriction fragments.


Studies of cloned sequences from four Drosophila heat shock loci. DNA cloned from the D. melanogaster (Oregon R) heat shock loci at 63BC and 95D codes for the 83,000 and the 68,000 dalton heat shock proteins, respectively. Both coding sequences occur once per haploid genome. Sequences complementary to messenger RNA for the 70,000 dalton heat shock protein are represented five times, twice at 87A and three times at 87 C. The copies at 87A differ characteristically from those at 87C in an interval of a few hundred bp near the 5' end of the messenger sequence, and the corresponding two classes of hsp 70 messenger RNA are found on polysomes after heat shock. Within this differential region, there is about 15% divergence between messenger sequences cloned from the two loci, while in the rest of the messenger region examined the homology is much closer although still imperfect. Unexpectedly, considerable homology is found between the sequence for the 68,000 dalton heat shock protein at 95D and the sequences for the 70,000 dalton protein at 87A and 87C, and between these sequences and a site in 87D. Messenger RNA molecules of 2.4, 2.55 and 3.05 kb code for the 68,000, 70,000 and 83,000 dalton heat shock proteins and hybridize to apparently uninterrupted DNA sequences of 2.1, 2.25 and 2.6 kb, respectively.


Clinical studies on cerebral schistosomiasis japonica in the Philippines. The author examined the cerebral schistosomiasis japonica (CSJ) in the Philippines which is one of the areas heavily infected with S. japonicum. Seventy-five subjects were selected randomly from 307 patients with CSJ, who showed neurological symptoms such as convulsions, paroxysmal disturbance of consciousness and hemiparesis. The mean age of the subjects was 33. Of the 71 patients who had paroxysmal disease, 54 had convulsions, in 33 of which it was of the Jacksonian type, and 24 had psychomotor seizures and 1 autonomic seizures. Thus, 58 patients or 82% of the paroxysmal disease group showed a sign of the localized lesion of the brain. Fifty-one patients (72%) of this group had attacks more than once a month, and the onset of the paroxysmal disease was later than 20 years old in 49 (69%). EEGs were judged as abnormal in 24 (32% of total subjects), borderline in 13 (17%) and normal in 38 (51%). The characteristic abnormal or borderline findings of EEG were random and paroxysmal slow waves with asymmetry. Discussion was made in reference to the strong suspicion that the cerebral symptoms of the subjects, the paroxysmal diseases in particular, were a syndrome associated with Schistosoma japonicum and to the difference between CSJ in Japan and that in the Philippines.


Proposal for a program in particle-beam radiation therapy in the the United States. A report from the Committee for Radiation Oncology Studies (CROS) and its Particle Subcommittee. The Program for Particle Therapy proposes utilization of hospital-based particle generators in a nationwide program to evaluate, through meaningful clinical trials, particle radiation therapy and the impact its utilization can have in cancer care. The scientific rationale for use of particle therapy compared to conventional radiation in the effort to achieve uncomplicated local control of cancer, to heal, cure and palliate the patient, indicates the advantages of particle therapy consist of either or both a) enhanced biological effect and b) physical properties leading to improvement in dose distribution. It has been estimated that in tho control local-regional cancer. Any new modality enabling the therapist to increase dose to tumor, while sparing critical normal tissue, can enhance local control and benefit systemic therapy. Limited clinical trials to date warrant further definitive clinical study of particle beams. Physical and biologic considerations of fast-neutron beams have been essentially completed; equipment design, availability, and predicted reliability are good; and the medical community has indicated support of further study. A major clinical investigation can be implemented to provide the scientific basis for judging clinical merit of use of high LET radiations. Concurrently, the first phase of work can be started with protons, negative pions, and heavy ions. It is anticipated that clinical results will accrue much more rapidly with hospital-based units for clinical trials; this Program proposes this transfer of particle technology from the laboratory to such hospital-based facilities in two phases, over a 10-year period.


[Effect of creatine and other phosphorylable amino compounds on the ileal absorption of Ca-45 in the rat]. Some carbohydrates which increase calcium absorption were phosphate acceptors. When administrated to adult Rat in ileal ligated loop, phosphorylable amino compounds such as creatine, L-aspartic and L-glutamic acids also increased calcium absorption; other effective compounds such as D- and L-lysine and D-alanine might be involved in reactions of phorphorylation. L-alanine, L- and D-valine and asparagine were ineffective in enhancing calcium absorption and were not phosphorylable. Injection of creatine into ileal loop induced the formation of its phosphorylated derivative. Absorption of the amino compound was not correlated with the fact that they were effective on calcium absorption.


[Effects of temperature on K, Na and water movements in isolated hepatocytes]. Isolated hepatocytes incubated at 1 degrees C loss K, gain Na and water. Rewarming results in a transient increase in K and Na permeabilities. These are insensitive to quinine, suggesting that they are not caused by an intracellular Ca accumulation.


[Demonstration of dopaminergic receptors in human pituitary adenomas secreting prolactin]. 3H Domperidone binding on cellular membranes from human prolactin adenomas demonstrates the presence of two dopaminergic binding sites. The mean value of the dissociation constant (Kd) for five adenomas is of 0.29 +/- 0.14 nM for the first site and of 4.19 +/- 1.56 nM for the second site. The maximal number of binding sites (Bmax) varies from one adenoma to another. The binding is completely displaced at 30 nM of tritiated Domperidone by apomorohine, a specific dopaminergic agonist.


[Isolation and composition of plasma membranes of bovine glomerulus]. Plasma membranes of Beef kidney glomeruli were prepared by ultracentrifugation on a discontinuous sucrose density gradient. They were characterized by specific marker enzyme activities and electron microscopy morphological studies. Amino acid composition revealed a high amount of hydrophobic residues. Carbohydrate amount was found to be as high as 8.9 p. cent of the dry weight.


[In vitro differentiation of primitive molar tissue from mouse embryo]. The developing molar tissues of the lower jaw of 13, 14 and 15 day Mouse embryos were removed and cultured in vitro. Complete molar dentition differentiated.


[The "hitch-hiking" effect of a selected gene and gamete combination (linkage disequilibrium): the example of 2 closely linked loci in the swine, Hal (halothane sensitivity) and PHI (phosphohexose isomerase)]. The observation of two lines of pigs selected differently for four generations confirms a recent theoretical work (Thomson, 1977) showing that a possible source of linkage disequilibrium may be the hitch-hiking effect of a selected locus (Hal) on another closely linked neutral locus (PHI).


[Heme structure in cooperative and noncooperative hemoglobins: study by resonant Raman diffusion]. The modifications of heme sites of hemoglobin, which should occur upon apoprotein alterations (responsible for variations of oxygen affinity), have been examined by Resonnant Raman scattering. The oxygenated (R) and deoxygenated (T) shape of apoprotein do not modify the heme states. The spectral differences between these forms are essentially due to the presence or the absence of the sixth ligand.


[Differential interaction of 2,4,6-trinitrobenzenesulfonic (TNBS) and 2,4-dinitrobenzenesulfonic (DNBS) acids on mouse lymphocytes]. Modifications of lymphoid cells with dinitro-2,4 phenylsulfonic acid (DNBS) or trinitro-2,4,6 phenylsulfonic acid (TNBS) have been studied. TNBS action always produces an electrophoretic mobility increase in relation with the amount of amino-groups, according to the cell type. DNBS action produces an electrophoretic mobility increase for B cells of spleen and a decrease for T cells of spleen and thymic cells. The hydrophobic fluorescent probe 1-anilino-8-naphthalene sulphonate used to study conformational changes of cells revealed a slight fluorescence decrease for TNP-modified cells and an important fluorescence increase for DNP-modified cells.


[Antigenicity of ribosomal proteins from a malignant mouse cell line. Preliminary results]. An original immunological two-dimensional technique with two different gels permits the protein analysis of 80 S cytoplasmic ribosomal murine particles and of their 60 S and 40 S subunits. The diagrams isolate and characterize a definite number of specific polypeptidic structures from each ribosomal particle.


[Fucosidosis and blood group substances in the urine]. Glycopeptide and oligosaccharide fractions obtained from fucosidosis urine contains more Lea activity (4-8 fold) than control urine. Both fucosidosis fractions also contained Leb and H activities, no A activity in contrast to the salivary and erythrocyte phenotypes (A, Le a+ b-). The amount of Leb activity is lower in both fractions (1) and (2) than that of Leb control children (4-16 fold decrease). Blood group A activity was not detected at any concentration used (less than or equal to 50 fold-concentrated urine) whereas A activity was found in (A, Le a- b+) control urine. On the contrary, both fucosidosis fractions contained H activity, whereas A (Le a- b+) control children fractions had none (less than or equal to 50-fold concentrated urine). H and Leb activity might originate from Lea precursor apart from the "non-secretory" type of the patient.


[Conformational coupling between structural units. A decisive step in the functional structure formation]. Proteins built up of several structural domains are compared with oligomeric proteins made up of several subunits. On this basis the functional consequences of a conformational coupling between domains are analyzed. From the data obtained on the elastase molecule it is proposed that the last step in protein folding insures the optimal coupling between domains and the energy of interaction is utilized for catalysis.


[Reduction of isoprenaline induced tachycardia in pregnant rats. Role of a serum factor]. The in vivo isoprenaline (50 microgram/kg) induced tachycardia (beta 1 adrenergic stimulant effect) was decreased in pregnant (20th day) Rats compared to non pregnant Rats. The in vitro positive chronotropic effect of isoprenaline (10 ng/ml) on cultured Rat heart cells was abolished by pregnant Rat serum whereas progesterone (0.10 to 5 microgram/ml) or oestradiol (0.1 to 25 microgram/ml) were ineffective. The decreased beta 1 adrenergic responsiveness in pregnant Rats could be related to a seric factor, different from these two hormones.


[Immunochemical study of protein anomalies in platelets of patients with Glanzmann's thrombasthenia and Bernard-Soulier syndrome]. Crossed immunoelectrophoresis of "Triton X-100" solubilized proteins from (125I) iodinated normal and abnormal (Bernard-Soulier and thrombasthenic) platelets was performed using rabbit antibodies raised against whole, normal platelets. In Bernard-Soulier platelets neither of the two surface oriented forms of GP I, glycocalicin and glycoprotein Ib (band 13) were detected. In two patients with type I thrombasthenia, the most prominent precipitate (No. 16) and platelet fibrinogen (No 24) were not detected whereas in one patient with type II thromblasthenia, fibrinogen was normally detected and the amount of protein 16 was around 15% of the normal level. The alloantibody IgG L..., able to induce a "thrombasthenic-like" reactivity in normal human platelets, has been shown to be directed against protein 16.


[Different primary structure of 2 variants of Friend virus p 30 polypeptide separated by isoelectric focusing]. Variants of p 30 (iso-p 30s), the 30,000 dalton major polypeptide of murine C-type retraviruses, have been characterized in all virus strains by isoelectric focusing. Several of these iso-p 30s have been found to coexist in a given virus strain. In the present study, two iso-p 30s, characteristic of the Friend-Rauscher subgroup, separated by preparative isoelectric focusing of p 30 in thin layers of polydextran gel, were subjected to tryptic peptide mapping and aminoacid analysis. The two iso-p 30s, with isoelectric points (pI) of 6.5 and 7.1, were found to possess a partially different primary structure, with about 50% homology. The results are discussed.


[Enzymatic activity of peritoneal macrophages of Balb C mice after enteral or parenteral stimulation with bacterial extracts]. A study of peritoneal macrophage activation has been conducted by simultaneous investigation in eleven enzymes in Balb/c Mice stimulated with bacterial extracts, by oral and parenteral route. The results showed important changes in Macrophage number and activity. On the other hand this activation depended upon the route of administration, antigen nature, and time interval between stimulation and peritoneal harvesting.


[Demonstration of measles virus antigens in osteoclasts in Paget's disease of bone]. The first results of histo-immunological studies on biopsies in Paget's bone disease strongly favour the presence of antigenic material of viral origin in osteoclasts. Measles virus may play a role in the etiology of Paget's bone disease.


[Isoprenaline, modifications of the turnover of ATP in the perfused rat heart]. 5 hrs after a single sub-cutaneous injection of the drug in a dose of 5 mg/kg, the ATP concentration was reduced by 15%, the incorporation of 14C adenine was augmented by 20% and the turnover of ATP was accelerated: the reduction of specific radioactivity attained 16%, 75 mn after the period of marquage, instead of 7% in the control hearts. When the drug was added to the perfusion fluid, there was no supplementary reduction of the ATP concentration and the action on the nucleotide turnover only existed if the ATP level was reduced by a pretreatment: the reduction of specific radioactivity then attained 24% after 75 min. of perfusion.


[Unmasking of the fast sodium channel in less than 4 day old embryonic chicken heart by inhibitors of sodium inactivation]. Embryonic Chick hearts aged less than 4 days are not always sensitive to tetrodotoxin, an inhibitor of fast sodium channel. It is shown that in the most frequent cases, in which tetrodotoxin sensitivity is apparently absent, this sensitivity can be demonstrated after previous treatment by veratridine or by toxin II of androctonus australis Hector Scorpion venom. It is concluded that the fast tetrodotoxin-sensitive sodium channel is regularly present in the heart of Chick embryos aged 2 and 3 days, but most often in a permanently inactivated state.


[Correlation between the regional blood volume and epileptic seizures in Papio papio]. The method of labelling red cells with technetium-99m was used to measured regional blood volume auring different types of epileptic seizures induced in the Baboon Papio papio. During seizures the cerebral blood volume increases and there is simultaneously a decrease of blood volume in nasal and hepatic regions, and a transitory increase of blood volume in the forepaws.


[Serotoninergic innervation of the paraventricular organ in chick embryo and chick; histofluorescence and autoradiography study optic and electron microscopy]. Fluorescence microscopy (Falck and Hillarp technique) of 11th day old chick embryo shows aminergic neurons and processes in the paraventricular organ area. Administration of tritiated amines permits us to observe uptake and storage of serotonin only as early as the 10th day; this suggests a very early hypothalamic serotoninergic nervous system differentiation.


[Protein kinase activity in Escherichia coli]. When growing E. coli in a minimal medium, at least four proteins from the soluble fraction and one ribosome-associated protein are found phosphorylated at the level of their threonine and serine residues.


[Morphological study of paramyxovirus Yucaipa]. Paramyxovirus Yucaipa exhibits a dramatic polymorphism. One can observe round particles, 250 nm in diameter, and numerous filamentous forms, some of which are 1800 nm long.


[Propionic acidemia and sodium dipropylacetate]. After administering sodium dipropylacetate to normal rats and biotin-deficient rats, we observed a slight hyperglycinemia, significant constant hyperglycinuria, but no accumulation of propionic acid in the blood or significant urinary elimination. Hepatic propionyl-CoA carboxylase activity, in vivo and in vitro, did not change, neither did sodium dipropylacetate inhibit propionyl-CoA carboxylase nor induce propionic acidemia resulting from a deficiency in this enzyme.


[Immunostimulating and adjuvant activities of a low molecular weight lipopeptide]. From crude extracts of a Streptomyces strain exhibiting immunopotentiating effects, a tetrapeptide was isolated and its structure established as L Ala leads to D isoGlu leads to L, L Dap comes from Gly. This peptide was devoid of biological activity but its chemical coupling with lauric acid gave a substance endowed with adjuvant and immunostimulating properties. This substance and the corresponding synthetic lauroyltetrapeptide were as active in this respect as the muramyl-dipeptide, thus far considered as the minimal adjuvant-active structure of bacterial cell walls: the presence of a sugar moiety is therefore not a prerequisite for immunopotentiating activities.


[Discovery and purification of a virus in gland hyperplasia of insects. Study of Merodon equistris F. (Diptera, Syrphidae)]. ADNA virus was purified from the salivary gland affected by a large proliferation in the Diptera Merodon equestris. Structure, nucleic acid as well as development and intracellular cycle of the virus have been characterized.


[Purification of the vascular plasminogen activator]. We have isolated a purified preparation of the vascular plasminogen activator. Plasma obtained from post occlusion venous blood has been chromatographed with low and high ionic strength buffers. The purified protein has a relative molecular weight of 71,000.


[Regulation of macronuclear DNA content during the cell cycle of the ciliate Tetrahymena paravorax (microstome form)]. Results of autoradiographic studies of 3H-thymidine incorporation support a model of macronuclear DNA content regulation involving an action upon the extremes of DNA content: elimination of S phase in cells with large DNA content, additional S phase in cells with DNA content. The frequency of each of these phenomena is about 20%: the inequality of frequencies obtained with the two types of sister cells (proters and opisthes) is relatable to the asymetry of cytodieresis observed in this ciliate.


[Effect of ovariectomy on circulating calcitonin levels in the rat]. Ovariectomy in 3 month-old Rats is followed by a fall in plasma CT level and a decreased secretion of CT after a calcium load. These results show that ovarian hormones may play a role in the direct or indirect control of the secretion of CT in the adult female.


[Effect of cell proliferation on the condensation of the X chromosome in culture medium of the individual karyotype 49 XXXXX]. The influence of cell proliferation on the condensation of the X chromosome was observed in vitro in human fibroblasts with 49 XXXXX karyotype. The frequency of cells with four Barr-bodies is low during the logarithmic growth phase and increases to 80% when the cells are becoming confluent or, independently of cell contact, when cell growth is arrested in a medium with low serum content. The condensation of th X chromosomes is reversible when the cells start growing again in medium with a higher serum content.


[Kinetics of the penetration of benzo(a)pyrene and benzo(k)fluoranthene into cells]. Microspectrofluorometric methods have been used to study the kinetics of the increase of fluorescence due to the intake of Benzo(k)Fluoranthène and Benzo(a)Pyrène in single living L cells. A computer program based on the methods of the least square fit has allowed the determination of the kinetic parameters. The results show that pinocytosis cannot be the unique mechanism of intake.