The organization, methods, and experience of a parenteral nutrition service. Malnutrition is an important component of prolonged illness. Parenteral nutrition can improve nutritional status, and its use is now common practice in certain disease states. In this paper the methods used by a parenteral nutrition service are described. The indications for, results from, and complications of, this form of therapy are also documented. This has been done on the basis of results obtained from a prospective survey of the use of parenteral nutrition in a major hospital over a one-year period.


The respiratory chain of Hydrogenomonas H16. The respiratory chain of Hydrogenomonas H16 is shown to consist of two pathways, one of which has as its oxidase and only cytochrome a high-potential b-type cytochrome which is concluded to be cytochrome o. The Km values for the oxygen of the two pathways are consistent with the high-potential b-type cytochrome functioning as a relatively high-affinity oxidase.


An enzyme-linked immunosorbent assay (ELISA) for detecting chicken anti-reovirus antibody at high sensitivity. An enzyme-linked immunosorbent assay (ELISA) with high sensitivity was developed for use in the avian system. Anti-reovirus antibody produced in chickens against the isolants S1133, UMI-203, Reo-25, and Winterfield were detected with this method at high titers (up to 1:14,000).


Chemotherapeutically acitve nitro compounds. 4. 5-Nitroimidazoles (Part III). More than 100 1-methyl-5-nitroimidazoles substituted in the 2-position via an oxymethyl group were synthesized and their structure-activity relationship toward various protozoa was investigated. Among the derivatives substituted with an aromatic radical there are most of the compounds which are highly effective against trichomonads; 9 preparations are superior to tinidazole and 29 are superior to metronidazole in mice infected with Trichomonas fetus, and 9 compounds exhibit a better effect than metronidazole in golden hamsters intrahepatically infected with Entamoeba histolytica. In the same series one dialkylamino-acetamide derivative shows excellent trypanocidal activity in the NMRI mouse, but this effect is limited to Trypanosoma brucei; 12 preparations developed a trypanocidal effect only after relatively high doses; their range of efficacy included Trypanosoma cruzi, among others, after repeated treatment. Of the carboxyl acid, carbamic acid and sulphonic acid esters synthesized only the already known group of carbamic acid esters possesses a pronounced antiprotozoal effect. Among the preparations substituted with a heterocyclic radical some of the pyridine derivatives proved to have distinct trichomonacidal activity. The influence of the type of substitution and the stability of the C-X bond in 2-substituted 5-nitroimidazoles of all compounds synthesized so far (I--III, 4th report) are discussed in 2 tables.


Medroxyprogesterone acetate: peripheral serum levels and its effects on pituitary and ovarian functions in the rhesus monkey. 17 alpha-Acetoxy-6 alpha-methylprogesterone (medroxy-progesterone acetate, MPA) in doses of 50 mg followed 4 weeks later by an additional 100 mg was i.m. injected in 5 adult normal female monkeys. Serum levels of MPA, progesterone, estradiol-17-beta and LH were determined every other day for a period of 8 weeks then weekly for 8 months. Peak values of serum MPA were detected 2 weeks postinjection followed by a gradual decline to undetectable levels after variable periods ranging from 125 to 225 days. The drug, in the doses used, caused inhibition of LH and estradiol surges and suppression of progesterone for more than 6 months. The return of menstrual cyclicity preceded the return of ovulatory cyclicity. Menstrual cyclicity occurred while the drug was still detectable in the blood. The probable site of action of the drug is discussed.


Intracytoplasmic type A particles from mammary tumours and leukaemias of strain ICRC mice. The ovarian-hormone-induced leukaemias of strain ICRC mice, with an abundance of intracytoplasmic type A particles in primary as well as transplanted lesions, were used to study morphological, biophysical, immunological and structural characteristics of type A particles. Mammary tumours of strains ICRC and C3H(Jax) were also used as sources for type A particles. The purified virions banded at the density of 1.20 g/ml in 12--60% linear sucrose-density gradient when subjected to spinning at 113,000 g for 4 h. The SDS-polyacrylamide-gel electrophoresis of type A particles from mammary tumours and leukaemias reproducibly resolved at least 8 polypeptides, 2 of these 54,000 and 24,000 dalton proteins, showing variable expression. Type A particles and B particles, despite the fact that each had a distinct polypeptide pattern, showed common antigens with different electrophoretic mobilities. Proteins of 24,000, 18,000 and 12,000 daltons from B particles were found to be antigenically related to those from type A particles. The bioassay studies carried out with purified A particles showed that 2/7 males of strain ICRC and 1/6 females of strain DBA-MTI developed leukaemias, as against none in the controls, when inoculated between the ages of 1-7 days. Spleen tumour and cervical tumour were seen in one female each of strain DBA-MTI.


Monocyte-induced increase in osmotic fragility of human red cells sensitized with anti-D alloantibodies. The mechanism by which human monocytes increase the osmotic fragility of red cells sensitized with Rhesus alloantibodies anti-D was studied in vitro. Both the increase in osmotic fragility and the lysis of red cells by monocytes were enhanced by cytochalasin B and were inhibited by hydrocortisone. These effects were similar to the effects of these agents on lysosomal enzyme release by monocytes. However, hydrocortisone was completely ineffective when added 1 h after mixing monocytes and sensitized red cells. This indicates that the damage responsible for the fragility increase and lysis is completed within 1 h and suggests that it is due to lysosomal enzymes released by the monocytes. Since for the full expression of the osmotic fragility increase and lysis an incubation time much longer than 1 h is required, it appears that the latter phenomena are the non-specific sequelae of damage inflicted upon the red cell by released lysosomal enzymes.


Some immunological investigations on antithrombin III 'Budapest'. Immunoelectrophoretic studies were carried out on plasma from a patient with an abnormal antithrombin III molecule (antithrombin III 'Budapest'). One-dimensional immunoelectrophoresis using a commercial antibody to antithrombin III (antibody to alpha2-AtIII) showed two precipitin peaks of alpha2-AtIII antigen. Two-dimensional immunoelectrophoresis showed two precipitin peaks, one with normal electrophoretic mobility, the other with increased electrophoretic mobility. It was shown that alpha2-AtIII antigen with normal mobility was identical to normal alpha2-AtIII, and that the alpha2-AtIII antigen with increased electrophoretic mobility was antigenically deficient, and appeared to be present in higher concentration than the normal alpha2-AtIII antigen. Although one-dimensional immunoelectrophoresis on plasma from the patient's son showed only one peak, two-dimensional immunoelectrophoresis revealed that the son also had two populations of alpha2-AtIII, one with normal mobility, the other with increased mobility. However, the alpha2-AtIII antigen with normal mobility appeared to be present in higher concentration than the alpha2-AtIII antigen with increased mobility. One- and two-dimensional immunoelectrophoresis using a different commercial antibody to alpha2-AtIII showed only one precipitin peak using the patient's plasma. The precipitin peak observed following two-deminsional immunoelectrophoresis was asymmetric and showed increased mobility.


An artificial 'haemophilic' plasma for one-stage factor-VIII assay. An 'artificial' plasma for one-stage factor-VIII assays is made by incubating human plasma with EDTA, to destroy factor VIII, and afterwards removing the anticoagulant by dialysis. Bovine factor V is then added to a given level. In the assay, contact activation is controlled by adding contact product. It was confirmed that factor-VIII activity was destroyed and that the EDTA was freely dialysable. The fibrinogen in the treated plasma clotted normally with thrombin. Likely variation in the factor-V activity was found not to be critical. The concentration of fibrinogen and other factors was adequate. Variation between batches was small. The artificial plasma yielded assay results closely comparable to haemophilic plasma in samples with factor-VIII activities in the range 0.01--20.0 iu/ml; the mean results in the artificial system were estimated to be 0.997 x those in haemophilic plasma, with a 95% confidence interval of 0.901--1.103. Biological variability in individual assays was smaller in the artificial system than when haemophilic plasma was used. Instability at the bench was more often detected in the artificial system than in haemophilic plasma assays, but the effect was eliminated from the results by obtaining duplicated readings in a balanced order.


Vasculature of the optic nerve in anencephaly. The optic nerves and globes obtained from 6 anencephalics were studied histologically and compared to normal specimens obtained from 4 stillborn infants as well as 1 case of septo-optic dysplasia. Special emphasis was placed on examination of the optic nerve, and it was found that an average of 48 vessels per high-power field were seen posterior to the lamina cribrosa in anencephalics. In contrast, control globes had an average of 12 vessels per high-power field. This suggested that an increase in the vasculature of the hypoplastic optic nerve is a characteristic feature of anencephalics.


Purification and some physicochemical properties of staphylococcal enterotoxin D. A method was developed for the isolation of staphylococcal enterotoxin D in highly purified form from cultures of Staphylococcus aureus strain 1151m. The method involves removal of the toxin from the culture supernatant fluid with the ion-exchange resin CG-50 followed by chromatography on carboxymethylcellulose (twice) and by gel filtration on Sephadex G-75 (twice). The purified toxin is homogeneous by polyacrylamide gel and sodium dodecyl sulfate-polyacrylamide gel electrophoresis and double gel diffusion tests. It is a simple, colorless, antigenic protein with an isoelectric point of 7.4 as determined by isoelectric focusing. Its molecular weight was determined to be 27 300 +/- 700 by molecular sieve chromatography on Sephadex G-100 and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Its serological activity is stable over a wide range of pH values (1.2--10.7). The enterotoxin consists of 236 amino acid residues and contains no free sulfhydryl groups. End-group analysis showed serine to be the NH2-terminal amino acid and lysine to be the COOH-terminal amino acid.


Properties of Escherichia coli 16S ribosomal ribonucleic acid treated with 4,5',8-trimethylpsoralen and light. 16S rRNA reacted with the furocoumarin 4,5',8-trimethylpsoralen (trioxsalen) and 360-nm light showed a number of chemical and physical differences from untreated RNA. After extensive irradiation, five molecules of trioxsalen were bound per molecule of RNA. The trioxsalen-treated RNA had an altered ultraviolet absorption spectrum and a distinctive fluorescence emission spectrum. The modified RNA was significantly more resistant to T1 ribonuclease digestion than was control RNA. Treated RNA, when mixed with purified ribosomal proteins, was not functional in the in vitro reconstitution of 30S subunits and yielded more slowly sedimenting particles which were inactive in protein synthesis assays. By contrast, 16S rRNA within the 30S subunit structure did not exhibit these changes when reacted with the same dose of trioxsalen and light, suggesting that the ribosomal proteins were effective in protecting the RNA from interaction with the drug.


RNA replication: required intermediates and the dissociation of template, product, and Q beta replicase. Replication complexes containing only one molecule of Q beta replicase and one strand of midivariant RNA (MDV-1 RNA) template were prepared by incubating the replicase with an excess of MDV-1 (-) RNA. In the presence of excess minus strands, these monoenzyme replication complexes were shown to synthesize essentially pure MDV-1 (+) RNA in both the first and second cycles of replication. When an equivalent concentration of mutant MDV-1 (-) RNA was added to this reaction before completion of the first cycle of replication, only wild-type MDV-1 (+) RNA was produced in the first cycle, but both mutant and wild-type MDV-1 (+) RNA were produced in the second cycle of replication. These results demonstrate that a monoenzyme complex is competent to synthesize RNA and, therefore, that a multienzyme replication complex is not a necessary intermediate of replication. The data also imply that after the completion of chain elongation, the product strand is released from the replication complex and that the template and the replicase then dissociate.


Isolation of Euglena gracilis chloroplast 5S ribosomal RNA and mapping the 5S rRNA gene on chloroplast DNA. Ribosomal RNA (5S) from Euglena gracilis chloroplasts was isolated by preparative electrophoresis, labeled in vitro with 125I, and hybridized to restriction nuclease fragments from chloroplast DNA or cloned chloroplast DNA segments. Euglena chloroplast 5S rRNA is encoded in the chloroplast genome. The coding region of 5S rRNA has been positioned within the 5.6 kilobase pair (kbp) repeat which also codes for 16S and 23S rRNA. There are three 5S rRNA genes on the 130-kbp genome. The order of RNAs within a single repeat is 16S-23S-5S. The organization and size of the Euglena chloroplast ribosomal repeat is very similar to the ribosomal RNA operons of Escherichia coli.


Effects of tetracyclines on aldosterone- and insulin-mediated Na+ transport in the toad urinary bladder. The effect of oxytetracycline and demethylchlortetracycline on aldosterone- and insulin-mediated Na+ transport (short-circuit current) were examined in toad urinary bladders mounted in modified Ussing chambers. Oxytetracycline had little or no effect on either basal or aldosterone-mediated Na+ transport. In contrast, demethylchlortetracycline markedly inhibited both basal and aldosterone-mediated Na+ transport. Furthermore, demethylchlortetracycline inhibited the aldosterone response significantly out of proportion to its effects on basal Na+ transport. Neither of the drugs had an effect on insulin-mediated Na+ transport. Consequently, the natriuresis observed in certain patients treated with demethylchlortetracyline may be related to drug-induced renal resistance to the effects of aldosterone.


The use of 60Co-EDTA as an extracellular marker in frog skin. 60Co-EDTA was tested as an extracellular marker in isolated epithelia of frog skin. It was found to be non-toxic for frog skin and 0.1 mM EDTA proved to be enough to saturate any adsorption sites. Comparing with [14C]inulin, 60Co-EDTA marks a slightly greater extracellular space and this volume is constant between 15 min and 2 h. Furthermore it is reproducible and its use simplifies the methodology of the determination of water and electrolyte contents in the cells.


A potential role for phospholipids in the regulation of 3-hydroxy-3-methylglutaryl-coenzyme A reductase in cultured C-6 glial cells. Effects of N,N-dimethylethanolamine. The relation of the activity of the microsomal enzyme, 3-hydroxy-3-methylglutaryl coenzyme A reductase, to cellular phospholipid composition was studied in C-6 glial cells. Phospholipid composition was perturbed by growth of cells in the naturally occurring amino alcohol, N,N-dimethylethanolamine. After growth of C-6 glia in 5 mM N,N-dimethylethanolamine for 24 h, reductase activity was diminished by 50%. A similar diminution in cholesterol synthesis was observed. This effect was not accompanied by any parallel change in cell growth, DNA synthesis, protein synthesis, fatty acid synthetase activity, or microsomal NADPH-cytochrome c reductase activity. The inhibition of reductase activity by N,N-dimethylethanolamine was prevented by the addition of equimolar concentrations of choline to the culture medium and, also, could be reversed completely by removal of N,N-dimethylethanolamine from the culture medium. The effect of N,N-dimethylethanolamine on reductase was associated with the formation of phosphatidyl-N,N-dimethylethanolamine which accumulated primarily at the expense of phosphatidylcholine and, after 24 h, accounted for 27% of total phospholipid phosphorus. The data demonstrate that incorporation of N,N-dimethylethanolamine into the polar head group of cellular phospholipids has a major impact on the regulation of the reductase. These observations may have particular relevance for the mechanisms of regulation of this enzyme, the cellular adaptation to alterations in membrane lipid composition, and the regulation of cholesterol synthesis in the developing nervous system.


[Radioimmunoassay of carbonic anhydrase I and II in rats. Application to the central nervous system during ontogeny]. A specific radioimmunoassay method for rat erythrocyte carbonic anhydrases I and II was developed using a double antibody system. Its sensitivity was in the nanogram range for each of the two isozymes. The method has been applied to the assay of cerebral carbonic anhydrase. Only CAII has been found in brain extracts of perfused rats. Accordingly, the assay of CAI in cerebral tissue can be used to quantify erythrocyte contamination on condition that the ratio CAII/CAI in blood had been worked out. The developmental change in the soluble and the Triton X-100 solubilized brain CAII from birth to adult is reported.


Modifications of aldolase during in vivo aging of rabbit red cells. Crude hemolysates, partially purified aldolase and aldolase purified to homogeneity from reticulocytes and mature erythrocytes, were incubated with a specific antiserum raised against crystalline rabbit muscle aldolase. We show that the same aldolasic activity corresponds to a greater amount of antigen in older than in younger cells, in crude hemolysates as well as in the above mentioned preparations; that is to say, old-cell aldolase contains cross-reacting material (CRM). Properties of purified enzyme from reticulocytes and mature erythrocytes were compared to those of muscle crystalline aldolase: -- the molecular specific activity of purified aldolase from erythrocytes is lower than with crystalline muscle aldolase, i.e. CRM is maintained throughout the purification steps. -- the specific activity of red cell aldolase towards both substrates (FDP and F1P) is lower than that of crystalline muscle aldolase. However, the ratio of activity towards the two substrates FDP/F1P is decreased in erythrocytes and reticulocytes. -- no other difference was found: Michaelis constant towards FDP, thermodenaturation constant and C terminal extremities are identical as are the molecular weights.


On the nature of tetracycline resistance in Bacillus subtilis mediated by the plasmid pT 127. The nature of tetracycline resistance was studied in a strain of Bacillus subtilis carrying the plasmid pT 127 in comparison with the parental strain. The resistance has been shown to be inducible in both strains upon exposure to subinhibitory concentrations of tetracycline. No modification of the protein-synthesizing activity of the ribosomes or intracellular inactivation of the antibiotic was observed in both strains. Accumulation of labeled tetracycline in B. subtilis was found to be particularly low in the wild-type strain, compared to other bacterial species, with concentration gradients of only 2 to 3 fold. From the kinetics obtained it is likely that the permeation of the antibiotic does not correspond to an active process in B. subtilis. A fairly good correlation was established between the level of resistance obtained after induction or by the presence of the plasmid pT 127 and a decrease in the binding capacity of the cell for the antibiotic.


[Some physico-chemical properties of the compact chromosome of Ananbaena variabilis]. It was shown that total DNA of the blue-green algae A. variabilis can be isolated as a compact particle with the sedimentation constant of 2700--2900S by lysis of the cells with the TritonX-100--sodium deoxycholate mixture in 1.0 M NaCl. The structure isolated is sensitive to ribonuclease and the hydrodynamic shift. It was found that within a compact chromosome A. variabilis DNA is circular and negatively superhelicized. The density of superturns of the DNA in 0,2 M NaCl at 20 degrees C is 0,061, which corresponds to one negative turn per 165 pairs of DNA bases.


[Effect of pancreatic DNAse on DNA synthesis in Bacillus subtilis]. An addition of pancreatic DNAse to the cultural medium is found to stimulate DNA synthesis and proliferation of Bacillus subtilis cells. Pancreatic DNAse induces a single-stranded disruption of Bac. subtilis DNA, which may act as a mechanism of DNA synthesis increase and of the culture growth acceleration.


[Phycobiliproteins of blue-green, red and cryptophytic algae]. The present-day concepts on phycobiliproteins, the protein pigments of blue-green, red and cryptophyte algae are reviewed. The functions, distribution, localization, physico-chemical, spectral and immunochemical properties of phycobiliproteins are described. The properties of the polypeptide protein subunits and the composition and chemical structure of chromophores as well as their binding to the apoprotein molecules are discussed.


[Cyanide-resistant respiration of Pseudomonas aeruginosa bacteria]. The transition of the bacterial culture into the stationary growth phase is accompanied by an appearance of cyanide-resistant respiration. Chloramphenicol inhibits the development of cyanide-resistant respiration. The cyanide-resistant oxidase is localized in the bacterial membrane. Its appearance is not due to the quantitative and qualitative changes of flavins, non-heme iron, ubiquinone and cytochromes of the b and c types, but is accompanied by an increase in the copper content of the membrane preparations. Neither cyanide-sensitive, nor cyanide-resistant chains of the bacterial electron transfer contain cytochromes of the a type. The cyanide-resistant oxidase accepts electrons at the ubiquinone--cytochrome b level of the main respiratory chain. The cyanide-resistant respiration is not accompanied by a formation of hydrogen peroxide. Cytochrome o performs the function of cyanide-sensitive oxidase. The nature of cyanide-resistant oxidase still remains obscure.


Urinary metabolites of 2-ethyl-2-methylsuccinimide (ethosuximide) studied by combined gas chromatography mass spectrometry. Metabolites of 2-ethyl-2-methylsuccinimide (ethosuximide) have been studied by combined gas chromatography mass spectrometry in a urine sample from a patient treated for petit mal epilepsy with ethosuximide. A new metabolite, 2-carboxymethyl-2-methylsuccinimide, was identified in the urine. It is presumably formed by omega 1-hydroxylation of the ethyl sidechain of the drug followed by a further oxidation of the primary alcohol to a carboxyl group. A previously identified metabolite, 2-ethyl-3-hydroxy-2-methylsuccinimide, was shown in the present study to yield two gas chromatographic peaks (as the N-methylated compound), indicating the existence of a diastereoisomeric pair. Thus, the enzyme responsible for the ring hydroxylation is probably not stereospecific, or alternatively two enzymes with different stereospecificity may exist.


Studies on composites of collagen and a synthetic polymer. Second report - mode of reaction of a laminar composite with living tissue, and results of long-term implantation. We have been studying composites of synthetic polymers and proctase-treated bovine collagen (telopeptide-poor collagen). In a previous paper, it is described that one of our composites appears to have a high tissue affinity, because it was found under a scanning electron microscope, well bonding the living tissue 6 weeks after implantation in the rabbit subcutaneous tissue. In the present study, the composites were also implanted in the subcutaneous tissue in rabbits to observe the mode of its bonding to the living tissue, and at the same time the bonding force was measured at various intervals during a period of 1.5 years, to observe the degree and duration of the effect of the composition, and further to study the optimal conditions for preparation of such a composite. As a method of composition, ultraviolet radiation and glutaraldehyde treatment were also comparatively examined, besides gamma-radiation that had been employed in a previous study. As the results, it is revealed that the composite bonds to the living tissue in such a manner that a portion of the collagen part is digested and absorbed to be replaced by the invading connective tissue and a part of collagen near the surface of the synthetic polymers had escaped digestion to combine with the rabbit's own collagen fiber. From these results we like to insist that the mode of reaction of our composites may be similar to real organization and be different in natur from mere encapsulation which was shown by the artificial materials hitherto in use to be considered to have a good tissue affinity. Therefor we think now the encapsulation is pseudoorganization. It is also shown that the bonding force of the composite to the living tissue and its duration depend chiefly on the degree of swelling, that is, the degree of intermolecular cross-linking, of the composed collagen part. Out of the gamma-radiated composites, those radiated 1--3 Mrads proved desirable in both tissue affinity and duration of tissue bonding, with a potent bonding force remaining effective even 1.5 years later, while the composite radiated with 5 Mrads or more proved poorer in these properties because of destruction of the collagen part, vastly varying in the properties. By ultraviolet radiation, it was easy to control the degree of swelling of collagen part with the radiation dose, however, this method gave a lower bonding force and a shorter duration of the force than gamma-radiation. The glutaraldehyde method allowed the collagen to remain over a long time, but gave the composite less tissue affinity than the other two methods.


Identification of amino sugars from bacterial lipopolysaccharides by gas chromatography electron impact and chemical ionization mass spectrometry. Amino sugars isolated from lipopolysaccharides of Brucella suis, Brucella abortus and Neisseria gonorrhoeae colony types 1 and 4 were identified using gas chromatography electron impact and chemical ionization mass spectrometry. Lipopolysaccharides were obtained by aqueous ether or aqueous phenol extraction. Isolated lipopolysaccharides were hydrolyzed in 1% acetic acid followed by hydrolysis of the polysaccharide moiety in 2 NHCl for 6 h at 100 degrees C. Amino sugars were first isolated by elution from Dowex 50 H+ and then N-acetylated, followed by trimethylsilylation. Trimethylsilyl ethers of 2-acetamido-2-deoxysugars; N-acetylglucosamine, N-acetylmannosamine, N-acetylgalactosamine, and a 2-acetamido-2.6-dideoxysugar, N-acetylquinovosamine, were identified by their fragmentation patterns. In the electron impact mode, N-acetylglucosamine and N-acetyl-galactosamine were distinguished from one another by comparing peak intensities at m/e 233 and 305. However, N-acetylglucosamine and N-acetylmannosamine could not be differentiated by electron impact mass spectrometry. In the chemical ionization mode, N-acetylglucosamine and N-acetylmannosamine both with base peaks at m/e 494, could be distinguished from N-acetylgalactosamine and N-acetylquinovosamine by their base peaks at m/e 420 and 332, respectively. N-Acetylglucosamine and N-acetylmannosamine were differentiated from one another by comparing peak intensities at m/e 330, 404, 420, and 510 [MH]+. This is the first report of chemical ionization mass spectrometry applied to the identification of amino sugars in bacterial lipopolysaccharides and shows that some 2-amino-2-deoxysugars can be differentiated by both electron impact and chemical ionization mass spectrometry.


Enzyme immobilization by radiation-induced polymerization of 2-hydroxyethyl methacrylate at low temperatures. Enzyme immobilization by radiation-induced polymerization of hydrophilic glass-forming monomers, such as 2-hydroxyethyl methacrylate, was studied. Enzyme radiation damage could be sufficiently retarded at low temperatures. The immobilized enzyme activity yield was markedly higher at low temperature than at higher temperature polymerization. At low temperatures the polymerized composite had a porous structure owing to ice crystallization which depends on the monomer concentration. It was deduced that the enzyme was partially trapped on the polymer surface, partially isolated in the pore, and partially occluded inside the polymer matrix. A decrease in activity caused by enzyme leakage was observed with repeated use in enzyme reactions where the composites had a large porosity. The activity yield showed a maximum at certain optimum porosities, i.e., at optimum monomer concentrations. Continuous enzyme reaction was preferably carried out using immobilized enzyme columns.


Fractionation of the proteolytic and amylolytic complex enzyme system of streptomyces aureofaciens and some properties of fractions. The Streptomyces aureofaciens extracellular proteolytic system was split into four fractions by carboxymethylcellulose (CMC) column chromatography giving three purely caseinolytic fractions and one fraction active toward both starch and casein. The first caseinolytic and amylolytic fraction was further fractionated by DEAE-Sephadex A-50 chromatography into one purely amylolytic fraction and another showing both activities, was refractioned into four new fractions by DEAE-cellulose chromatography. These fractions were found to be heterogeneous by polyacrylamide gel electrophoresis, three of them acted on both starch and casein and a fourth was only caseinolytic. The second CMC fraction was further purified by CMC rechromatography to an homogeneous fraction that hydrolyzes carboxypeptidase A(EC 3.4.2.1) synthetic substrates and solubilizes elastin. It had only one polypeptide chain with a molecular weight of about 28000 daltons, a high thermal stability in the presence of calcium ions, a pH optimum of about 6.8, and a maximal caseinolytic activity at about 50 degrees C.


[Ultrastructural aspects of preglioma].

[Ultrastructural aspects of preglioma]. Investigation of submicroscopic changes in the astrocytes after the intracerebral injection of the carcinogen 9,10-dimethyl-1-2-benzanthracene suggested that the so-called substantial period of preglioma began from the 45th day of the experiment, when structurally atypical astrocytes appeared against the background of distrophic changes. The changes in the submicroscopic organization of the astrocytes during chemical carcinogenesis can be divided into three stages: 1) intracellular hyperplasia due to traumatic injury of the brain; 2) dystrophic changes connected with the vascular tissue disturbances and immediate action of the carcinogen on the tissue; 3) atypical ultrastructure reconstruction of the astrocytes.


In vitro inhibition of digestive enzymes by heavy metals and their reversal by chelating agent: Part I. Mercuric chloride intoxication. The effect of mercury on alkaline phosphatase, lipase, aminotripeptidase and glycylglycine dipeptidase in the liver and digestive tract of Channa punctatus is investigated in vitro. Mercury inhibits the activities of all these enzymes and the degree of inhibition increases with the increase in the concentration of the metal. Addition of EDTA, a chelating agent, restored the mercury inhibited enzyme activity and the degree of restoration was related to the concentration of the chelating agent.


In vitro inhibition of digestive enzymes by heavy metals and their reversal by chelating agent: Part II. Lead nitrate intoxication. The effect of lead on alkaline phosphatase, lipase, aminotripeptidase and glycylglycine dipeptidase in the liver and digestive tract of Channa punctatus is investigated in vitro. Mercury inhibits the activities of all these enzymes and the degree of inhibition increased with the increase in the concentration of the metal. Addition of EDTA, a chelating agent, restored the mercury inhibited enzyme activity and the degree of restoration was related to the concentration of the chelating agent.


The fixed horizontal neutron therapy beam at Edinburgh: dosimetry and radiation protection. A compact cyclotron producing 15 MeV deuterons has been installed at the Western General Hospital, Edinburgh, to extend the Medical Research Council's clinical trials of fast neutrons. Two treatment rooms are available one of which has an isocentric unit and the other a fixed horizontal beam which is the subject of this paper. A radiation protection survey has demonstrated safe levels of radiation throughout the building although there is some activation of the fixed horizontal beam cone resulting in doses to radiographers of 10 mrem per week. The calibration of neutron dose is discussed and the measurements of dose distribution described. Isodose and depth-dose data for both the neutron and photon components of the field are presented.


Enzymes involved in the degradation of thyrotropin releasing hormone (TRH) and luteinizing hormone releasing hormone (LH-RH) in bovine brain. As part of an investigation of neuropeptide inactivation mechanisms, we have resolved an enzymatic activity in bovine brain which catalyzes the deamidation of thyrotropin releasing hormone (TRH) and the hydrolysis of the Pro9--Gly10--NH2 bond of luteinizing hormone releasing hormone (LH-RH) from a second LH-RH degrading activity which does not degrade TRH. The former activity is similar, if not identical to, the post-proline cleaving enzyme in kidney as it is active toward the post-proline cleaving enzyme substrate CbzGly--Pro--Leu--Gly and inhibited by CbzPro--Phe and diisopropylfluorophosphate. In addition, products derived from the degradation of TRH and LH-RH by this activity show a specific cleavage on the carboxyl side of a proline residue. The latter activity has not yet been characterized with respect to its site of cleavage of the LH-RH molecule due to the presence of other contaminating peptidases.


Localization of prostaglandin synthetase in chicken epiphyseal cartilage. Prostaglandin synthetase activity in high-speed particulate fractions of chick epiphyseal cartilage has been characterized with respect to cofactor requirements, pH optimum, buffer-ion effects, types of prostaglandins formed, and the distribution of prostaglandin synthetase activity in zones of the epiphyseal plate. Direct homogenization of cartilage was found to be more efficacious than releasing chondrocytes by enzymatic digestion for preparation of prostaglandin synthetase, a homogenization time of 4 min yielding maximal activity. The optimal incubation medium contained 50 mM Tris buffer (pH 7.5), 2.5 mM epinephrine, 1 micronM hemoglobin, 3.25 mM glutathione, 200 microgram/ml enzyme protein, and 5 micronM substrate. Glutathione was effective only if present during homogenization. Rates of PGE2 biosynthesis were linear up to 15 min and then rapidly declined, indicative of self-deactivation. The low levels of PGF2alpha formed, and their decrease after 20 min incubation, suggests the possible presence of degradative enzymes. Prostaglandin synthetase was inhibited by aspirin, indomethacin, and vitamin E, but not vitamin K1. Cation concentrations in the physiological range had only modest effects on prostaglandin biosynthesis, and then only if present during tissue homogenization. In the presence of phosphate buffer, Ca2+ was somewhat inhibitory. Since in the absence of phosphate Ca2+ had no deleterious effect, it is probably that the inhibitory effect was caused by precipitation of calcium phosphate. Hypertrophic and calcified cartilage exhibited significantly higher prostaglandin synthetase activity than the proliferating and maturing zones. The increased synthesis of prostaglandins in the low layers of the growth plate may indicate a role of these factors in chondrocyte differentiation and/or calcification.


Characterization of an octopamine-sensitive adenylate cyclase from insect brain (Mamestra configurata Wlk.). An adenylate cyclase present in the brain of the moth Mamestra configurata Wlk. that is stimulated selectively by low (micromolar) concentrations of octopamine has been characterized with respect to several properties. The optimum pH, optimum ATP:Mg2+ ratio, the concentration of ATP required for half-maximal and maximal reaction velocity, metal ion specificity, effect of NaF, and effects of GTP and 5'-guanylylimidodiphosphate were in general similar to those of catecholamine-sensitive adenylate cyclases from various regions of mammalian brain. However, ethylene glycol bis-(beta-aminoethyl ether)-N,N-tetraacetic acid (EGTA), a calcium chelator, stimulated both basal and octopamine-sensitive enzyme activity in the insect brain, whereas in mammalian brain EGTA is usually observed to inhibit basal activity but not catecholamine-stimulated activity. Adenylate cyclase activity of the 47,000 g particulate fraction of the insect brain was almost undetectable in the absence of added GTP. Addition of saturating concentrations (100 micrometer) of GTP to the particles restored about 30% of the basal and octopamine-sensitive enzyme activity present in the homogenate. Addition of 100,000 g supernatant to the particles doubled both basal and octopamine-sensitive enzyme activity in the presence of saturating concentrations of GTP, indicating that in addition to GTP, a cytosolic factor(s) is necessary for enhanced adenylate cyclase activity.


A rapid method for the purification of antibody--enzyme conjugates. A method is described for a rapid and efficient separation of enzyme-labelled antibodies from the free enzyme following the coupling reaction. A single passage of the reaction mixture on a protein A--Sepharose CL-4B column gave a sharp separation of the free enzyme from the conjugate.


Strategic withdrawal from cigarette smoking. Cigarette smoking can be viewed mainly as a psychological habit with accompanying pharmacological and social satisfactions and reinforcements. It is mainly a learned behavior, occurring in response to direct and indirect social pressures. Though each person's smoking habit is individualized, studies show that smokers find at least six psychological satisfactions in the habit: stimulation, handling, relaxation, tension reduction, craving and habit. To quit smoking, the individual must undergo physical and psychological withdrawal. Physical withdrawal symptoms last about a week but it may take months or years to restructure a life without cigarettes. Quitting is a long-term learning process involving a great deal of unconscious, constructive and painful conflict. One can stop smoking if the problem is attacked on a broad front, if one "burns one's bridges" behind him/her and if suitable satisfying alternatives and involvements are discovered. Physical exercise in the form of aerobics is proposed as an especially effective alternative behavior.


Transport of Neisseria meningitidis cultures: growth-supporting media and freezing as an alternative. The abilities of Transgrow (TG), Thayer-Martin (TM), and New York City (NYC) solid media to maintain the viability of 12 strains of Neisseria meningitidis under various controlled conditions were assessed. The effects of charcoal impregnation of swabs, temperature, and an enriched CO2 atmosphere were examined with holding for up to 21 days. Recovery from samples held at 35 degrees C was, in almost all instances, greater than at 22 or 4 degrees C. A strong requirement for added CO2 was demonstrated, especially at lower temperatures. No positive effect could be attributed to the use of charcoal-impregnated swabs. NYC and TM media were the best overall, with the former permitting recovery from more than 75% of all samples held on slants for 20 days at 4 degrees C in an atmosphere of 5% CO2 in air. Freezing, with holding on dry ice, was a useful alternative to the use of growth-supporting media. This latter method eliminated the requirement for an enriched CO2 atmosphere.


Induction of reaginic (IgE) gonococcal antibodies in the rat by a common antigen of Neisseria gonorrhoeae. An antigen (ZAB) common to Neisseria gonorrhoeae was prepared by stepwise elution of a crude gonococcal antigen (ZA) from columns of diethylaminoethyl cellulose employing 0.02 M phosphate buffers, pH 7.6, containing increasing concentrations of sodium chloride. Rats immunized with ZAB produced reaginic (IgE) antibody which cross-reacted with ZA prepared from eight gonococcal strains by the passive cutaneous anaphylaxis (PCA) test. Heating of the sera at 56 degrees C for 4 h destroyed the PCA activity. The PCA activity of the anti-ZAB rat serum was removed after absorption with ZAB antigen or with rabbit anti-rat IgE but not after absorption with gonococcal lipopolysaccharide or with heat-killed or formalinized gonococci. Treatment of ZAB with trypsin or heating at 100 degrees C for 30 min destroyed or reduced the antigenic activity respectively. Further purification of ZAB by filtration through Sephadex G-100 gave a preparation (ZAB2) which contained the common antigen as shown by the cross-reactivity of anti-ZAB2 rat serum with seven stains of N. gonorrhoeae. Fraction ZAB2 contained material which had a molecular weight less than 13,700 and was associated with the presence of material absorbing at 260 nm. The results of this study indicate that a low molecular weight antigen, which appears to be protein in nature and associated with nuclei acid, is common to the gonococcus and is the main antigenic component inducing reaginic (IgE) antibody in the rat.


Hydroxylase regulation in Candida tropicalis grown on alkanes. Candida tropicalis synthesizes a hydroxylase (3 to 5 nmol of product formed per minute per milligram of protein) and a cytochrome P-450 (0.10 to 0.13 nmol per milligram of protein) during growth on n-tetradecane. A three- to four-fold increase in the level of NADPH cytochrome c reductase is also observed in those cells as compared to the level of cells grown on glycerol. The most efficient inducers of the hydroxylase and of cytochrome P-450 are straight-chain alkanes having at least 10 carbon atoms. Alkenes and higher alcohols are also good inducers. There is little or no growth on ramified hydrocarbons such as pristane and on long-chain aldehydes and fatty acids. The partial inhibition of growth on decane is probably due to the denaturation of the microsomal electron carrier systems by the fatty acid formed by hydroxylation of the decane in the yeast.


The importance of the "bay region" diol-epoxide in 7,12-dimethylbenz[a]anthracene skin tumor initiation and mutagenesis. The skin tumor-initiating and V79 mutagenic activities of various derivatives of 7,12-dimethylbenz[a]anthracene (DMBA) were investigated to determine what possible cellular metabolite(s) may be responsible for its carcinogenicity and/or mutagenicity. 1-,2-,3-,4- and 5-hydroxyDMBA were found to be essentially inactive as skin tumor initiators whereas 9- and 10-hydroxyDMBA had weak activity. The (+/-)-trans DMBA 8,9- and 5,6-dihydrodiols were also essentially inactive as skin tumor initiators and (+/-)-DMBA 8beta,9alpha-diol-10alpha-11alpha-epoxide had weak skin tumor initiating activity. All of the above tested derivatives of DMBA were essentially inactive as mutagens in the cell-mediated or direct V79 mutagenesis systems. A methyl or fluoro addition to the 1, 2 or 5 positions almost completely blocked the skin tumor initiating and V79 mutagenic activities of DMBA, whereas a fluoro addition to position 11 did not. From our data we suggest that a 'bay region' diol-epoxide may be important in DMBA carcinogenicity and mutagenicity.


DNA repair in Syrian hamster embryo cells treated with 7,12-dimethylbenz[a]anthracene and its weakly carcinogenic 5-fluoro analog. The postreplication repair capacity of Syrian hamster embryo cells in culture was determined after treatment with the potent carcinogen 7,12-dimethylbenz[a]anthracene (DMBA) and its weakly carcinogenic analog 5-F-DMBA. The size and amount of daughter DNA sedimenting as high-molecular-weight DNA were found to be less in the DMBA treated cells than in the 5-F-DMBA-treated cells. This difference probably depends upon the types of adducts entering DNA replication.


Comparative metabolism and DNA binding of 7,12-dimethylbenz[a]anthracene and its weakly carcinogenic 5-fluoro analog. Cultured Syrian hamster embryo cells readily convert both the carcinogenic polycyclic aromatic hydrocarbon, 7,12-dimethylbenz[a]anthracene (DMBA), and its weakly carcinogenic analogue, 5-fluoro-7,12-dimethylbenz[a]anthracene (5F-DMBA), to water soluble metabolites. DMBA, however, binds to the hamster cell DNA at least 2.8-3.0 times more extensively than 5F-DMBA. Likewise, 5F-DMBA is converted to water soluble metabolites by liver microsomal preparations at a rate equal to DMBA, but the latter compound binds to DNA 2.6-3.2 times more effectively.


Changes in microtubule packing during the stretching of an extensible microtubule bundle in the ciliate Nassula. The cytopharyngeal sheath in the ciliate Nassula is a long hollow tube-shaped microtubule bundle that forms part of a large feeding organelle called the cytopharyngeal basket. During the initial stages of ingestion of algal filaments by Nassula the sheath is stretched, becomes approximately elliptical in cross-section, and its external cross-sectional perimeter increased by a factor of about two. The mean circumferential centre-to-centre spacing of radially oriented rows of sheath tubules increases from 57 to 137 nm during stretching but sheath thickness and the radial spacing of sheath tubules do not change appreciably. It is suggested that extensible circumferentially oriented intertubule links and relatively inextensible radial links may define the anisometric mechanical properties of this particular microtubule bundle which are related to its cytoskeletal role. The possibility that extensible links resist stretching elastically and provide the restoring forces for return of the sheath to its former shape and dimensions after stretching is considered.


[Retrograde inhibitory effect of continuous reinstillation of digestive juice in temporary artificial and pathological enterostomies on the secretory digestive flow]. The reinstillation of the digestive juice in the lower end of an enterostomy has a retrograde inhibitory effect on the upper digestive secretions (average inhibition of 30,2%) particularly on the intestinal juice during the digestive syndromes associated with one or several interruptions of the continuity of the bovel. It should be noted, however, that the observation has not been made on normal men, but rather on severely ill patients treated in the intensive care unit of a gastro-intestinal disease department.


[Stimulation of G-6-P independent glycogen synthase activity in the human placenta]. In immature human placentas, the activity of the I (glucose-6-phosphate-independent) form of glycogen synthase is significantly increased by insulin, glucose and by both compounds associated. In full-term placentas, the same kind of results has been found in each organ studied; due to the great variability observed in synthetase I control activities, the stimulation is significant only in the presence of insulin.


[New data on the biology of gametocytes of Plasmodium yoelii yoelli gathered from morphological characteristics indicating their age]. A morphological study has been made of the gametocytes of P. yoelii (a parasite of Rodents belonging to the "vivax" group). The authors distinguish 4 types chronologically ordered. Moreover, morphology enables one to predict their infectivity. The morphological analysis represents a tool leading to new data on the biology of gametocytes: loss of infectivity as soon as the parasitaemia becomes high; infectivity restricted to the young stages; concentration of young stages in the blood ingested by the Mosquitoes; short periodicity and short lived infectivity.


[Isolated rat hepatocytes. Simultaneous study of variations in sialic acid content of glycoconjugated membranes and asialotransferrin uptake]. Hepatocytes isolated from streptozotocin treated Rats bind less asialotransferrin than hepatocytes isolated from normal rats. This decrease is parallel with a decrease in the sialic acid content. Insulin therapy restored simultaneously membrane sialic acid content and asialotransferrin binding capacity.


[Stimulation of proliferation of rat spleen cells, in vitro, by a nonspecific acute inflammatory exudate. Modulation of cell response to phytohemagglutinin (PHA)]. The effect of an acute non-specific inflammatory exudate with mitogenic activity on macrophages in culture has been tested on the spontaneous and PHA-induced DNA synthesis of spleen cells in vitro. Stimulatory effect of this exudate was observed on spontaneous DNA synthesis which was detectable over a range of 1 : 4 to 1 : 4,000 concentrations. After optimal PHA stimulation, an inhibition of mitogen-induced DNA synthesis was observed when the cells were exposed to the highest concentrations (up to 1 : 128) of the exudate. Thereafter, the phenomenon could be reversed and the stimulation was maximal at a concentration of 1 : 2,000. When a sub-optimal dose of PHA was used, the simulatory effect was more pronounced and detected from 1 : 8 up to 1 : 4,000 concentrations.


[In vitro biosynthesis of beta-lipotropin in brain tissue]. We have recently developed methods to identify biosynthetized beta-endorphin and beta-lipotropin (beta-LPH) following incubation of Rat pars intermedia with radioactive amino acids. We used the same approach for rat brain tissue. In the striatum we found a peptide similar to beta-LPH while its identification in hypothalamus was less positive. This is the first demonstration of such biosynthesis and it could well be an important step in determining the biosynthetic patterns of cerebral endorphins and enkephalins.


[Demonstration of N-acetylglucosaminyl-transferases in the outer membrane of liver mitochondria]. Outer mitochondrial membranes catalyze the transfer of N-acetyl-glucosamine into polyprenic and proteinic endogenous acceptors. Different effectors (nucleotides, EGTA) are used, suggesting that there are two separate metabolic ways.


[Antiarrhythmic properties of 3-(2-hydroxy-3 isopropylamino-propoxy)-3 phenyl-2 isoindolinone-1, (RS, SR) in the dog]. In the Dog, 3-(2-hydroxy-3 isopropylamino-proxy)-2-phenyl-1-isoindolinone (RS, SR) possesses an anti-arrhythmic activity similar to that of quinidine but at dose levels 2 to 6 times lower than in the case of the latter compound. Furthermore, in contrast to quinidine, at the dose levels where the antiarrhythmic activity is well observed, the compound is devoid of hypotensive activity and of depressive action on cardiac contractility. The first clinical studies of this compound have shown its usefulness in the treatment of ventricular and supraventricular arrhythmias.


[Hemagglutination by simian adenovirus 7]. Simian adenovirus 7, either complete virus or its capsid subunits, agglutinates Rat (Sprague-Dowley) red blood cells in the presence of heterotypical antiserum. Haemagglutination takes place at 4 degrees C and room temperature. The antigen could not be eluted and its haemagglutinin properties are heat-stable. The reaction is specific. It is inhibited by homologous antiserum only. This property and its characteristics permit a camparison of this strongly oncogenic adenovirus to the human adenovirus of subgroup III of Rosen.


[Description of an unusual enzymic activity cleaving the third component of complement]. A large enzymatic complex cleaving C3 (C3'ase) in the absence of magnesium (EDTA) has been partially characterized in two patients. On gel filtration EDTA-C3'ase was found in an 800 000 fraction containing antigenic C4, IgA and IgG in addition to alpha-2-macroglobulin and IgM normally present. EDTA-C3'ase was specifically neutralized by antibodies to IgG, IgA and C4. The characteristics of this unusual enzymatic complex are compatible with that of a C4b2a complex stabilized by its specific binding to an auto-antibody of the IgG and/or IgA class.


[Phospholipid antigens acquired by the young forms of Schistosoma mansoni]. Immunofluorescence studies provide evidence of cardiolipin fixation at the schistosomulum's surface, following incubation with liposomes (cardiolipin-lecithin or cardiolipin-lecithin added with cholesterol). Fixation occurs at 37 degrees C as well as at 0 degree C whether proteins were present or not. Several washes do not remove cardiolipin fixation.


[Lipid thesaurismosis rheumatism. Arthropathies in alpha galactosidase A deficiency (Fabry's disease). Ultrastructural study of the synovial membrane demonstrating microcrystals in the mitochondria of synoviocytes]. Articular manifestations may be the onset of genetic alpha galactosidase deficiency (Fabry's disease). Ultrastructural study shows typical osmiophilic lamellar inclusions of trihexosylceramides in synoviocytes, capillaries and adipocytes. Furthermore microcrystals identical to those seen in Gaucher's disease and type II hyperlipoproteinemia were observed in mitochondria and free in cytoplasm. These data suggest a microcrystalline pathogenesis of these arthropathies, as in gout and chondrocalcinosis, and what we have generally called crystallopathic arthropathies.


Isolated gonococcal pulmonary valve endocarditis: diagnosis by echocardiography. The perplexing clinical course of a 23-year-old black male with isolated gonococcal pulmonary valvular endocarditis is presented. M-mode echocardiography provided the first clue to the presence of pulmonary valvular vegetations and the proper diagnosis. Since Neisseria gonorrhea appears to have a particular affinity for the pulmonary valve, the presence of isolated pulmonary valvular endocarditis should raise the strong possibility that Neisseria gonorrhea is the offending organism. This case report of pulmonary valvular vegetations detected by echocardiography strongly emphasizes that all four cardiac valves must be visualized in order to rule out the presence of echocardiographically detectable valvular vegetations.


Substrate-labeled fluorescent immunoassay for phenytoin in human serum. A homogeneous substrate-labeled fluorescent immunoassay has been applied to the measurement of phenytoin concentrations in human serum. We coupled a fluorogenic enzyme substrate, galactosyl-umbelliferone, covalently to a derivative of phenytoin. Under assay conditions, this drug-substrate conjugate was nonfluorescent but became fluorescent upon hydrolysis catalyzed by bacterial beta-galactosidase. When antibody to phenytoin is bound to the drug-substrate conjugate, it is inactive as an enzyme substrate. Addition of phenytoin to competitive-binding reactions relieves the inactivation, and the resulting fluorescence is proportional to the phenytoin concentration. We validated the fluorescent immunoassay by comparing values for phenytoin obtained with this technique to those obtained by gas chromatography and by enzyme immunoassay (EMIT). All three methods correlated well. The major metabolite of phenytoin, 5-(p-hydroxyphenyl)-5-phenylhydantoin, and other drugs at concentrations expected in serum had no effect on the assay. The fluorescent immunoassay is rapid and simple to perform and requires only 2 microL of serum sample per test.


Interference of iodine-125 ligands in radioimmunoassay: evidence implicating thyroxine-binding globulin. Although there is abundant published evidence that radioiodinated antigens interfere in digoxin radioimmunoassays, other radioimmunoassays are similarly affected. We investigated the relationship of radioiodinated antigen structure to its function in the immunoassay. Carrier-free 125I-labeled iodotyrosine and iodohistamine derivatives were incubated with human serum, and the bound and free fractions were separated. We demonstrated that diiodotyrosyl analogs bind avidly to serum proteins. Because protein binding could be reduced with competitors that inhibit thyroxine-binding globulin, such as 1,8-anilinonaphthalene sulfonate and thyroxine, thyroxine-binding globulin was clearly implicated. Diiodotyrosyl compounds also bound to solutions of purified thyroxine-binding globulin, and this binding was inhibited by the same two competitors. We postulate that thyroxine-binding globulin is the major source of the heretofore unexplained interference of radioiodinated haptens. We present recommendations for eliminating or minimizing such interference.


Immunoelectrophoretic studies on human small intestinal brush border proteins. A quantitative study of brush border enzymes from single small intestinal biopsies. A method for measuring brush border membrane enzymes from small intestinal biopsies by crossed immunoelectrophoresis is presented. The use of a brush border specific antiserum made isolation of the brush border membrane before analysis unnecessary. This prevented loss of material which, together with inactivation of enzymes, was a limiting factor in previous studies of brush border enzymes from peroral biopsies. In 58 biopsies from patients without gastrointestinal disorders a close correlation between antigenic activity and corresponding enzymatic activity was shown for the following enzymes: sucrase-isomaltase (EC 3.2.1.48-EC 3.2.1.10), lactase-phlorizin hydrolase (EC 3.2.1.23-EC 3.2.1.62), microvillus aminopeptidase (microsomal, EC 3.4.11.2) and dipeptidyl peptidase IV (EC 3.4.14.X). The immunoelectrophoretic patterns of intestinal mucosa near the ligament of Treitz, and in jejunum and ileum were established. The method presented is thought to be of value in further studies of the molecular basis of brush border diseases.


Carbonic anhydrase isoenzyme B in erythrocytes of subjects with chronic acidosis. A specific and quantitative immunological method for determination of human erythrocyte carbonic anhydrase isoenzyme B has been used to ascertain the contents of this enzyme in the erythrocytes of healthy persons and of subjects with chronic metabolic and respiratory acidosis. The investigations have shown significant increase of carbonic anhydrase type B in the erythrocytes of patients suffering from renal failure with chronic acidosis, and in patients with chronic obstructive lung disease and chronic respiratory acidosis. The erythrocytes of acidotic uremic patients have a significantly higher content of erythrocyte carbonic anhydrase isoenzyme B than do the erythrocytes of uremic subjects without chronic acidosis. In chronic obstructive lung disease, the content of this enzyme in erythrocytes was significantly higher in the hypercapnic patients than in the normocapnic ones. In renal failure, significant correlation was found between carbonic anhydrase isoenzyme B and standard bicarbonate. In chronic obstructive lung disease no significant correlation was found between carbonic anhydrase and pCO2.


Effect of zinc supplementation on plasma levels of vitamin A and retinol-binding protein in malnourished children. The relationship between plasma levels of vitamin A and zinc was studied in 45 children suffering from vitamin A deficiency and 20 children with protein-energy malnutrition. Thirty apparently normal children of the same age group were also studied for comparison. The mean levels of plasma vitamin A, retinol-binding protein and zinc were significantly lower in vitamin A-deficient children and in children with PEM, as compared to controls. Supplementation with 40 mg zinc daily for 5--10 days resulted in a significant increase in plasma vitamin A and RBP levels in children with PEM but not in the vitamin A-deficient group. There was, however, no correlation between plasma levels of vitamin A and zinc. The data suggest that in children with PEM, apart from deficiencies of protein and vitamin A, zinc deficiency may also contribute to the lowering of plasma vitamin A levels. They also suggest that in vitamin A-deficient children, without protein-energy malnutrition, zinc deficiency does not seem to have a role.


Absence of oroticaciduria in adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency. Orotic acid excretion was normal when tested by three methods in adenosine deaminase deficiency and purine nucleoside phosphorylase deficiency. These results do not support the speculation, based on the oroticaciduria observed by others, that the immunodeficiency in these disorders results from the inhibition of pyrimidine biosynthesis. An alternative hypothesis is discussed.


Circulating carcinoembryonic antigen immune complexes in sera of patients with carcinomata of the gastrointestinal tract. The sera of patients with histologically proven carcinomata of the gastrointestinal tract were fractionated by gel filtration and the fractions assayed for the presence of free carcinoembryonic antigen (CEA) binding immunoglobulins and CEA immune complexes by radioimmuno-double-diffusion, using 125I-CEA as a marker. In eleven out of thirteen cases with disease recurrence, the presence of CEA-IgM complexes was observed, and in three out of thirteen cases the presence of CEA-IgG complexes, could be demonstrated. Free CEA-binding immunoglobulins could not be detected.


Opioids: similarity between evaluations of subjective effects and animal self-administration results. Abuse potential studies of 33 morphine-like analgesics were compared in humans and monkeys. The results of intravenous self-administration studies in rhesus monkeys were correlated with measures of morphine-like signs, symptoms, and subjective effects in ex-addicts. Each set of data was assigned to a position in a 3 x 3 contingency table dependent upon whether the results were yes, no, or equivocal. Of the 33 drugs, 29 were given identical classifications in both the human and animal test procedures. This good concordance between the human and animal results further validates each procedure and suggests the possibility that both the human and animal procedures are measuring a common underlying pharmacological property which relates to abuse potential of drugs.


Biochemical and clinical effects of ethane-1-hydroxy-1,1-diphosphonate in calcium nephrolithiasis. 1. The short- and longer-term effects of ethane-1-hydroxy-1,1-diphosphonate (EHDP), an inhibitor of crystal growth and potential preventive agent against urinary tract stones in man, have been studied. 2. Measurement of urinary excretion of EHDP was used to define the best dosage regimen. When 4.4 mmol of EHDP mmol of EHDP was given in four divided doses the murinary concentration of EHDP achieved was high enough (10-5 mol/1) to inhibit the crystallization of calcium crystals throughout the day. 3. Nine patients with recurrent calcium stones were given this dose of EHDP daily for 12 months and seven were then studied for a further 12 months under placebo. During treatment with EHDP, inhibitory activity in urine towards precipitation of calcium phosphate was restored from low values to greatly above normal. This could be accounted for by the inhibitory effect of EHDP itself, coupled with an increase in urinary inorganic pyrophosphate. After stopping EHDP the excretion of EHDP rapidly fell to undetectable levels but the excretion of pryophosphate remained elevated throughout the 12 months of placebo treatment. EHDP also induced a rise in plasma phosphate and an increase in the urinary excretion of oxalic acid and uric acid, but these changes were all fully reversible when EHDP was stopped. 4. The average rate of stone formation per patient per year decreased from 2.4 to 0.2 during treatment with EHDP and remained low during the following 24 months. However, the dose needed for this effect is known to affect bone turnover and mineralization.


Immunogenecity of hapten-protein conjugates in rabbits and monkeys preimmunized against the carrier protein. The effect of preimmunization with carrier protein on the subsequent response to immunization with hapten-protein conjugate was investigated. Rabbits immunized against bovine serum albumin (BSA) showed a delay in the production of anti-progesterone antibodies upon immunization with progesterone-11-BSA. After a booster injection with P-11-BSA, however, they achieved antiprogesterone levels comparable to those in animals immunized with P-11-BSA only. Similarly, rhesus monkeys preimmunized with tetanus toxoid (TT) showed a delay in the development of anti-hCG titers when immunized with beta-hCG-TT. Here again, after booster immunizations they achieved anti-hCG levels comparable to those of the control animals.


Dislodgment of pacemaker electrode simulating focal motor seizure. The unusual finding of dislocation of the electrode of a pacemaker in a patient caused repetitive twitching of the right leg. Initial confusion with focal motor seizure resulted in the administration of antiepileptic medication. Malposition of the electrode was confirmed by appropriate roentgenographic studies, and subsequent repositioning terminated the episode.


Ascending aorta-supraceliac abdominal aorta bypass: successful removal of an infected graft in the descending thoracic aorta. An infected graft and a mycotic pseudoaneurysm were successfully resected by employing an ascending aortasupraceliac abdominal aorta bypass graft in a 19-year-old man. He had formerly undergone graft replacement surgery for traumatic aneurysm of the descending thoracic aorta, with the aid of a temporary external bypass graft. After this first operation, the patient had suffered from septicemia due to Psudomonas aeruginosa, which resulted in formation of mycotic pseudoaneurysms at the distal anastomotic site of the prosthetic graft and at both stumps of the formerly employed external bypass graft.


[The action of nitroglycerine on digitalis induced ST depression in patients with coronary disease (author's transl)]. The influence of therapeutic digitalisation on ST depression due to myocardial ischemia was investigated in 11 patients, average age 53.6 years, with coronary heart disease, compared with the effectiveness of nitroglycerine. Therapeutic digitalis led to an average increase of ischaemic ST depression from -0.53 to -0.73 mV. The mean pulmonary arterial and pulmonary capillary pressure decreased slightly, the frequency of pectanginous attacks increased. Independent of the digitalis effect nitroglycerine had an opposing action on these parameters. In decompensated patients with coronary heart disease (n = 4) both digitalis and nitroglycerine produced a shift of the left ventricular function curve as an expression of improved cardiac action. This could not be observed in patients with compensated ventricular function (n = 7). In sufficient ventricular function digitalis led to a further increase of myocardial ischaemic ST depression. In ventricular insufficiency no uniform behaviour was apparent. ST depression induced by digitalis could be reversibly influenced by nitroglycerine.


[Cross allergy between sulphonamide diuretics, probenecid, sulphamethoxazole and sulphonyl-urea compounds (author's transl)]. Typical signs and symptoms of type III allergy occurred in a 55-year-old diabetic during diabetes and diuretic treatment with glisoxepid, glibenclamide, furosemide and probenecid. Symptoms of this type of allergy unintentionally recurred with every subsequent therapeutic administration of each one of these drugs. The possibility of a cross allergy between sulphonamide diuretics, chemotherapeutic sulphonamides and sulphonyl-urea compounds has only rarely been described.


[Treatment of decompensated valvular disease with nitroglycerin (author's transl)]. The effects of parenteral nitroglycerin after acute and continuous infusion were investigated in 12 patients with mitral and (or) aortic valvular disease (stage IV of the New York Heart Association) and severe therapy-resistant pulmonary congestion. Intravenous injection of 1 mg led to immediate and marked decrease of right atrial mean pressure, and pulmonary artery and pulmonary capillary mean pressures, whereas mean arterial blood pressure, stroke volume index, cardiac frequency, and cardiac index remained unchanged. With a dosage of 3-10 mg/h the pressure lowering of the right circulation could be sustained. Pressure lowering of the right circulation abolished pulmonary congestion and led to marked reduction of shortness of breath. The principle of venous pooling can thus not only be used successfully in cases of increased pulmonary capillary pressure due to primary myocardial insufficiency, but also in cases with pulmonary congestion due to decompensated valvular disease.


[Nitroglycerine in chronic obstructive pulmonary disease (author's transl)]. The effect of 0.8 mg nitroglycerine on haemodynamics and pulmonary functions was examined in 12 patients with chronic obstructive pulmonary disease. Resting heart rate was increased from 72 to 82/min by nitroglycerine, but at rest and on exercise there was only a slight reduction in systolic blood pressure, the diastolic level remaining essentially unchanged. There was a decrease in cardiac index of 16 and 15% at rest and on exercise, respectively. A highly insignificant reduction in mean pulmonary artery pressure occurred both at rest and on exercise. Respiratory variations of pulmonary artery pressure markedly decreased after nitroglycerine and correlated well with airway resistance, which fell by 28%. Pulmonary vascular resistance fell from 188 to 149 dyn.s.cm-5. Vital capacity increased after nitroglycerine as a sign of reduction of pulmonary emphysema. Partial pressure of arterial oxygen and carbon dioxide showed no definite changes at unchanged respiratory rates. Thus nitroglycerine produces a marked improvement in haemodynamics and pulmonary function without any increase in ventilatory distribution abnormalities. These results support the use of nitroglycerine as one of several means of treating chronic obstructive pulmonary disease.


[Effect of a combination of 3 pesticides on the sensitization of guinea pigs to ovalbumin]. The author treated guinea pigs with a combination of three pesticides: First group -- with cyneb in a dose of 12,5 mg/kg of body weight (threshold value) +/- sevine in a dose of Ip 5 mg/kg (twice threshold value) +/- tribufon in a dose of 5.0 mg/kg (threshold value); second group -- cyneb 125 mg/kg (ten times of threshold value) +/- sevine 1.5 mg/kg (twice the threshold value) +/- tribufon 5.0 mg/kg (threshold value). Two groups of animals were used as controls: first group-- treated with 2,0 mg/kg of cyclophosphamide and second group -- with 3% of starch emulsion, with which other substances were given. The indicated substances were administered in the animals by a probe (a catheter) daily, in the morning before feeding, for a period of six months. Then sensibilization was made by ovalbumin in a solution of 10 gm% according the scheme: first injection of 2 ml of ovalbumin -- 1 ml subcutaneously in the inguinal region and 1 ml intraperitonealy. A challenging dose of 2 ml was administered inraperitonealy after 20 days and the reaction of the experimental animals was examined for a period of 90 minutes after the administration of the challenging dose. There was no difference in the reaction of the guinea pigs, treated with a combination of three pesticides in comparison with animals, given the same pesticides in the same dosage, but singly.


Generalized epilepsy with bilateral synchronous spike and wave discharge. New findings concerning its physiological mechanisms. A hypothesis for the mechanism of generalized spike and wave discharge in human generalized epilepsy is proposed in the light of findings obtained in feline generalized penicillin epilepsy. It is postulated that generalized bilaterally synchronous spike and wave discharge depends upon a diffuse and relatively mild state of cortical hyperexcitability which increases the responsiveness of cortical neurons. Afferent thalamo-cortical volleys normally involved in the genesis of spindles and recruiting responses are most likely to precipitate spike and wave discharges under these conditions. The spike and wave pattern probably results from the activation of a recurrent intracortical inhibitory pathway which becomes activated when cortical neurons discharge in greater number and more repetitively than is normally the case. During spike and wave discharges a large number of neurons oscillate between short periods of excitation, corresponding to the spike, and longer periods of inhibition, corresponding to the slow wave component of the spike and wave complex. This disrupts the normal transactional processes of cortical neurons which are presumably responsible for mental activity, particularly for the close integration of perception, cognition and voluntary motor responsiveness. The degree of this interference varies greatly and in mild absence seizure it is not justified to speak of "loss of consciousness". The fundamental disturbance in absence seizures brought about by the generalized cortical spike and wave discharges is therefore better regarded as a "clouding of the mind". Loss of consciousness can be said to occur only when the interference with mental activity becomes particularly intense. Loss of consciousness in absence seizures can therefore not be used as an argument in favor of primary involvement of higher brain-stem mechanisms.


Evoked potential studies in petit mal epilepsy. Visual information processing in relation to spike and wave discharges. The evoked potential has been useful in studying characteristics of the epileptic nervous system. To investigate whether an alteration in sensory processing is possibly related to behavioral unresponsiveness observed to precede spike and wave bursts, average visual evoked potentials (VEPs) were studied during a 4.0 sec preburst interval in 6 patients with petit mal epilepsy. Individual responses, evoked by photic flashes, were averaged as a function of flash-to-burst interval. In 3 subjects with classical bursts of spike and wave, the average VEP was degraded 0.5 sec or less before burst onset. In 3 subjects with atypical electroencephalographic seizure discharge, VEP degradation was not seen. Some possible interpretations concerning the significance of the VEP alterations were discussed.


Movement epilepsy in the monkey with an experimental motor focus. In the monkey with an alumina focus in the primary motor area, proprioceptive afferents are crucial in triggering or modulating seizures. The reversible exclusion of these afferents by curarization results, depending on the level of activity of the focus, either in regularization of spontaneous seizure frequency and duration (recorded from the cortex), or in their subtotal suppression. In the last case, muscular stimulation is still able to elicit seizures. The triggering modality is highly specific: if it is possible to induce seizures with acoustic or cutaneous stimuli in normal conditions, this is no longer possible under curare. It may then be assumed that exteroceptive stimulation is effective through the startle reaction or flexion reflex that they elicit; in other words, the actual trigger would take origin in the indirect involvement of proprioceptive afferents. Physiological mechanisms and relations with human clinical features are discussed.


Involvement of the cerebellum in the initiation of fast ballistic movement in the monkey. Monkeys were trained to perform a fast ballistic flexion or extension of the elbow in response to a sound cue. The reaction time was about 250 msec in the normal monkey and remained so after deafferentation of the operant arm. In a cerebellectomized animal, the reaction time was consistently longer, being, on the average, 400 msec. This was associated with an equal retardation in the onset of neural changes in the motor cortex. These results are interpreted as evidence for the involvement of the cerebellum in the initiation of some fast ballistic movements.


Cortex, basal ganglia and cerebellum in motor control. For voluntary movement, function generators are necessary that are located in the brain-stem, cerebellum and basal ganglia. Following lesions of these generators, voluntary movements are impaired while stimulus-dependent movements are still possible. This discussion also applies to speech production. The motor function generators cooperate with the whole cerebral cortex since both tactical adaptation to the environment and strategic guidance by the motivation system contribute to voluntary action. The motor cortex plays an epicritical role, adding advanced tactile and proprioceptive guidance for those movements that need this kind of regulation, especially the fine finger movements which depend entirely on the motor cortex. The complexity of the cerebral potentials preceding voluntary movement corresponds to the activity of several motor subsystems acting in concert.


Muscular dystrophy contrasted with denervation: different mechanisms underlying spontaneous fibrillations. The data reviewed in this paper indicate that spontaneous fibrillations do not involve a cholinergic mechanism since non-depolarizing anticholinesterase drugs such as Mestinon fail to increase spontaneous fibrillations in denervated muscle. Fibrillation potentials are related to the changes in electrical properties of the membrane of denervated muscle fibres which lead to the appearance of spontaneous subthreshold depolarizations, sometimes triggering a propagated potential. Fibrillations seem to appear in cycles and this may depend on the depression of spontaneous depolarization by muscle activity itself. Fibrillations are also an important feature of Duchenne muscular dystrophy and polymyositis, but they have not been found in Landouzy-Dejerine muscular dystrophy. These "myopathic" fibrillations probably arise from subthreshold depolarizations in the membrane of muscle fibre segments which have been functionally or anatomically isolated from the end-plate by a pathological lesion (Fig. 4). Experimental demonstration of spontaneous fibrillations in baboon biceps muscles after extrajunctional myotomies indicates that such an isolated muscle fibre segment can indeed develop and sustain spontaneous fibrillation activities. Studies of motor unit potentials in myopathies by "coherent" electromyography disclose linked potentials after the main potential in Duchenne dystrophy, but not in Landouzy-Dejerine muscular dystrophy. The linked potentials are signs of collateral innervation by sprouts of the motor axons. The fact that linked potentials occur in Duchenne dystrophy, including in obviously dystrophic motor units (Fig. 5), shows that such motor axons are quite healthy and able to sprout efficiently. The muscle fibres thus innervated collaterally are probably the ones which fibrillated and were deprived of trophic motor control as a result of myopathic lesions of the type considered in Fig.4. This correlation receives support from the finding that both spontaneous fibrillations and linked potentials are lacking in Landouzy-Dejerine muscular dystrophy, which obviously presents a different type of muscle lesion.


Tests of the predictive value of EEG recording from within the brain in the partial epilepsies. The EEG, as recorded from electrodes in deep structures of the brain in cases of intractable seizures, has been surveyed to assess its predictive value in the partial epilepsies. In addition to scalp EEGs and depth EEGs in control situations, ictal recordings of tape-recorded seizures and the interictal enhancement of EEG signs by benzodiazepines have been analyzed. All results have been quantified by computer analysis. The predictive value of the EEGs has been assessed in 2 ways: (1) by comparison with the pathology found after surgery and (2) by the long-term clinical status of the patient postoperatively. Twelve cases with lesions found in limbic structures and 6 with cortical involvement are reported. In the latter cases secondary firing of the hippocampus, with its low threshold for seizures, frequently followed, though more rapidly than could be timed by visual inspection alone. Bilateral signs both in scalp EEGs and in depth were a common finding in the cases with limbic lesions. In those patients with marked clinical improvement, follow-up computerized studies of pre- and post-operative scalp EEGs revealed normalization in both hemispheres, leading to the proposal that a focus of defective tissue may impair neuronal function in widely distant regions of the brain.


Plasma levels of vitellogenin in Chrysemys picta during the annual gonadal cycle: measurement by specific radioimmunoassay. A RIA for turtle (Chrysemys picta) vitellogenin is described. After dimethylformamide precipitation of vitellogenin from the plasma of estrogen-treated female turtles, antibodies were developed in rabbits. The dimethylformamide precipitate was further purified by o-triethylaminoethyl cellulose column chromatography; the vitellogenin component eluted as a single peak. This material was used for iodination by a mild chloramine method. Antibodies to turtle vitellogenin did not cross-react with plasma from male turtles or vitellogenic females of other vertebrate groups, including lizards and snakes. Limited cross-reactivity exists among the chelonians, however. Using a 1:5000 dilution of antiserum, the limit of detection was 15 ng, and the midrange was 320 +/- 45 ng. For an antiserum dilution of 1:1000, these figures were 30 and 600 +/- 37 ng, respectively. Using this assay, the seasonal pattern of plasma vitellogenin in the turtle has been described, and preliminary studies on in vitro hepatic vitellogenesis have been performed.


Thyrotropin-releasing hormone stimulates release of arginine vasopressin and oxytocin in vivo. The effects of TRH upon neurohypophyseal hormone release were studied in conscious rabbits. Intravenous infusion of 250 nm/kg TRH had no significant effect on either arginine vasopressin (AVP) or oxytocin (OT) release, but a 5-fold greater dose led to significant increases in plasma levels of both AVP and OT and behavioral arousal. Intraventricular injection of 3 nm TRH produced significant elevations of both plasma AVP and OT, with even greater effects on behavior than after iv infusion. The maximal hormone response to intraventricular injection was observed considerably earlier than that for iv injection and the response occurred after an almost 1000-fold lower dose of TRH. Neither artificial cerebrospinal fluid vehicle nor the inactive analogue D-tyrosine2 TRH (p-Glu-d-Tyr-Proamide) had any effect on neurohypophyseal hormone release or on behavior. MK-771 [L-N-(2-oxopiperidin-6-YL-carbonyl)-L-histidyl-L-thiazolidine-4-carboxamide], a TRH analog with enhanced central nervous system effects, had effects on AVP and OT release comparable to equimolar doses of TRH. TRH stimulates release of both AVP and OT after both intraventricular and iv injection, and these effects may be independent of behavioral activation.


Seasonal variation and the influence of body temperature on plasma concentrations and binding of thyroxine and triiodothyronine in the woodchuck. Woodchuck plasma was collected during four seasons of the year and assayed for total and dialyzable (free) T4 and T3 and for rT3. Plasma concentrations of total and free T4 and T3 were higher in the spring (T4, 5.4 +/- 0.6 microgram/dl; free T4, 3.0 +/- 0.4 ng/dl; T3, 202 +/- 22 ng/dl; free T3, 0.51 +/- 0.04 ng/dl) and lower in the prehibernatory fattening period in summer (T4, 2.3 +/- 1.0 microgram/dl; free T4, 1.2 +/- 0.5 ng/dl; T3, 45 +/- 27 ng/dl; free T3, 0.16 +/- 0.10 ng/dl) and fall (T4, 3.2 +/- 1.0 microgram/dl; free T4, 1.3 +/- 0.2 ng/dl; T3, 130 +/- 12 ng/dl; free T3, 0.25 +/- 0.02 ng/dl). In spite of the extremely high concentrations of T3 in the winter (437 +/- 32 ng/dl), free T3 concentrations (0.034 +/- 0.003 ng/dl), when measured at the appropriate temperature for hibernation, were significantly lower than those found at other seasons of the year. Plasma binding of T3 was lower during the summer and increased again to approximately double the spring value during the winter. rT3 was at or below the sensitivity of the method (6 ng/dl) at all seasons. It is suggested that the wide seasonal variations in thyroid hormone concentrations and altered plasma protein binding may represent important adaptations influencing the metabolic rate and the process of hibernation in the woodchuck.


Prolactin concentrations in the monkey fetus during the last third of gestation. PRL concentrations were measured in cord plasma obtained at hysterotomy from 26 rhesus monkey fetuses between 111--170 days gestational age (GA). Mean PRL concentrations increased significantly from 23.7 +/- 10.1 (X +/- SE) ng/ml at 121--130 days GA to 126.9 +/- 16.9 ng/ml at 161--170 days GA. A similar significant increase in PRL with age also was observed in samples obtained from 16 fetuses chronically catheterized in utero between 130--155 days GA. Mean PRL levels were 34 +/- 3.2 ng/ml at 131--140 days GA and rose to 82 +/- 9.7 at 150--155 days GA. No difference in PRL concentrations was found between cord blood samples and fetal peripheral blood samples at the ages studied. Maternal PRL levels did not change in samples obtained from chronically catheterized, chair-restrained mothers between 130--155 days GA. A tendency toward an increase in maternal PRL with advancing gestational age was observed in samples collected after hysterotomy. These data indicate that the fetal rhesus monkey demonstrates an increase in plasma PRL similar to that in the human, suggesting a possible physiological role for this hormone in the primate fetus late in gestation.


Serum prolactin responses to TRH in recurrent breast cancer patients. The response of serum prolactin (PRL) to thyrotropin-releasing hormone (TRH) was evaluated by radioimmunoassay in 6 normal women and 44 breast cancer cases. They were divided into the following 5 groups: group 1:6 normal women; group 2:10 preoperative patients with early breast cancer; group 3:13 preoperative patients with advanced cancer; group 4:13 postoperative patients with no recurrence of cancer for more than 2 years; group 5:8 postoperative patients with cancer recurrence. The maximum increment of serum PRL levels following the administration of TRH was significantly higher in groups 2, 3 and 5 than in groups 1 and 4. These results indicate that patients with recurrent breast cancer have a higher PRL response to TRH than those without recurrence of cancer.


Location and encapsidation of the coat protein cistron of tobacco mosaic virus. A bidirectional elongation of the nucleoprotein rod. The coat protein cistron of tobacco mosaic virus has been located on the viral RNA starting between 975 and 1050 nucleotides from the 3'-hydroxyl end. This locates its 5' end close to the origin for virus assembly, where the first protein disk interacts with RNA. It also means that the coat protein mRNA must have a short 5'-untranslated tail and a long (over 500 nucleotides) 3' one. The recovery of characteristic oligonucleotides in nuclease-protected rods during the growth from RNA and a protein disk preparation shows that elongation of the nucleated rods proceeds independently in both directions though, on average, much more rapidly along the longer 5' tail than the shorter 3' tail. Protected RNA of length equal to that in the complete virion is first seen within 6 min, showing that the most rapidly elongated particles are substantially complete by this time.


Acyl-coenzyme-A synthetase I from Candida lipolytica. Purification, properties and immunochemical studies. Acyl-coenzyme-A synthetase I from Candida lipolytica has been purified to homogeneity as evidenced by polyacrylamide gel electrophoresis in the presence and absence of dodecylsulfate as well as by Ouchterlony double-diffusion analysis. The purification procedure involves resolution of cellular particles with Triton X-100 and chromatography on phosphocellulose, Blue-Sepharose and Sephadex G-100. The purified enzyme exhibits a specific activity of 20--24 U/mg protein at 25 degree C, which is about 100-fold higher than those of long-chain acyl-CoA synthetases hitherto reported. The molecular weight of the enzyme has been estimated by polyacrylamide gel electrophoresis in the presence of dodecylsulfate to be approximately 84 000. The enzyme is specific for fatty acids with 14--18 carbon atoms regardless of the degree of unsaturation. Studies with the use of specific antibody to acyl-CoA synthetase I have indicated that this enzyme is immunochemically distinguishable from acyl-CoA synthetase II.


The enzyme-bound copper of dopamine beta-monooxygenase. Reaction with copper chelators, preparation of the apoprotein, and kinetics of the reconstitution by added copper. The enzyme-bound copper of dopamine beta-monooxygenase reacted rapidly with the chelator bathocuproine disulfonate; the reaction in the presence of ascorbate was completed in 2 min at 25 degrees C with 1mM chelator. This reaction and also the reaction with EDTA could be used to prepare the apoenzyme, which in both cases was completely reactivated in less than 10 s. The reactivation data gave apparent Michaelis constants for copper 0.03 -- 0.2 micron. Trace amounts of copper in buffers and assay mixtures gave significant reactivation without added copper, unless they had been treated with a chelating resin. Titrations using the different chelation rates of free and enzyme-bound copper indicated that four copper atoms are bound per enzyme molecule of four subunits. The native enzyme was more stable against thermal inactivation than the apoenzyme, but this stability was only partially restored by addition of copper to the apoenzyme.


Iron absorption in patients with polycythemia vera: a comparative study using the whole-body counter and the ferrous sulfate absorption test. The absorption of iron given per os to ten polycythemic patients was examined by two methods: first by giving radioactive iron (59Fe) and utilizing a whole-body counter, and second by peroral administration of ferrous sulfate. A correlation of 80% was achieved by the two methods. A comparison was made between the patterns of the ferrous sulfate absorption in the polycythemic patients and in five patients with low iron serum due to bleeding hemorrhoids. No difference was observed between these two groups, with the exception of the initial values which were lower in the polycythemic patients. On the other hand, the absorption of iron was higher in polycythemic patients, indicating that serum iron level is not the only factor regulating iron absorption.


A whole body counter with an invariant response for whole body analysis. A scanning 6-detector whole body counter is described. Its characteristics include good sensitivity, high energy resolution, good counting efficiency, geometric uniformity and localisation information of incorporated radioactivity using collimators. Problems such as band-width, calibration for air and phantom and measurements on patients, are discussed. The whole body counter is compared to other systems.


Evaluation of linear profile scans; mathematical treatment of line-spread functions. The profile measurement capability of a scanning multidetector whole-body counter is described using line spread functions. Measurements in air and water phantoms, as a function of energy, are discussed in relationship to the relevant physical processes. A computer program for evaluating bodyprofile scans and an attempt to improve the resolution are reported.


Sanfilippo type C disease: clinical findings in four patients with a new variant of mucopolysaccharidosis III. A new genetic variant of the Sanfilippo syndrome due to deficiency of acetyl CoA: alpha-glucosaminide N-acetyltransferase, was recently demonstrated in four patients. The clinical findings of these patients are reported here. Differential diagnosis from other types of the Sanfilippo syndrome on clinical and routine laboratory criteria is difficult and enzyme assay is necessary to reach the diagnosis. Since two of the patients reported are females and consanguinity was present in one case, autosomal recessive inheritance is most probable.


Serologic distinction between the rabbit kappa L chain allotype b4 and an allele b4. Rabbit kappa light (L) chain allotypes are controlled by the allelic genes b4, b5, b6, b9 and a recently described allele b4v. The product of the b4v gene differs from that of b4 by at least two amino acid substitutions in the constant region. A serologic distinction between the b4 and b4v allotyes has now been detected by a radioimmune assay using one of six anti-b4 sera tested. The distinction was enhanced by fractionation of this antiserum on an immunoadsorbent containing cottontail rabbit IgG. IgG with L chains of the b4v type are deficient in their ability to inhibit binding of b4 IgG to the antibody preparation. L chains isolated from b4 and b4v IgG samples could not be distinguished by this assay indicating that interaction with an H chain is required for expression of the L chain determinants that differentiate b4 from b4v.


Exclusion of VHa and VHy loci expression on individual B cells from normal and VH allotype-suppressed rabbits. The distribution of two heavy chain subgroups, VHa and VHy, on rabbit peripheral blood lymphocytes was examined by double membrane immunofluorescence. Fluroescent anti-a1 and anti-y33 were found to react with separate B cell populations; no doubly stained cells were observed. Further evidence for the independent expression of genes controlling the VHa and VHy subgroups were obtained by neonatal suppression of a 2 or y33 in a2y33/a3y- heterozygous rabbits. Suppression of VHa did not affect the expression of VHy, nor did the suppression of VHy affect the expression of VHa. The expression of a single VH gene per B cell is in marked contrast to the simultaneous expression of multiple CH genes.


Interaction of anti-staphylococcal protein A antisera with Fc receptor-bearing human normal lymphocytes. Staphylococcal protein A (SpA) is known to bind the Fc region of IgG of most mammalians and to possess biologic activity both in vivo and in vitro, where it acts as a lymphocyte polyclonal mitogen. Its binding to the Fc gamma portion bears many features of the antibody-antigen interaction, such as the dissociation constant, lattice formation, and complement activation. Moreover, SpA seems to compete with membrane Fc receptors for IgG so that the possibility of an interaction with the same CH domain(s) of IgG can be considered. In the present study, evidence is given that anti-SpA antisera obtained from chickens and rabbits are able to inhibit EA rosette formation by normal human lymphocytes and that they are able to recognize, with immunofluorescent staining, a subpopulation of normal human peripheral blood lymphocytes (PBL) that closely resembles that of EA rosette-forming cells (RFC). Moreover, the depletion of EA RFC by means of a single gradient centrifugation is accomplished by the parallel depletion of PBL stainable by anti-SpA antisera. The relevance of these results in the hypothesis of a similarity between the combining sites of SpA and membrane Fc receptor(s) for IgG is discussed.


Anti-V region framework antibodies affect the ligand binding of VL dimer. The effect of antibodies to the light chain variable region (VL) of protein MOPC-315 (alpha, lambda 2), on the binding of hapten by VL315 dimer or Fv315 (VL + VH) was studied by equilibrium dialysis. Anti-VL did not change the binding properties of Fv but affected the binding properties of VL dimer. At pH 5, the binding properties of VL in the presence or absence of anti-VL were the same, whereas at pH 8, anti-VL reduced the number of ligands bound to VL from two to one. It has previously been shown that VL dimer binds one ligand at pH 5 and two ligands at pH 8, and that VL conformation at pH 5 is tighter. Hence, our results suggest that anti-VL tightens the conformation of VL dimer at pH 8.0 such that it can bind only one ligend. Since Fv is not affected by anti-VL, the results indicate that a combining site made of two identical chains (VL dimer) can undergo a conformational change upon interaction with its antibody. Such conformational change can indirectly affect the binding properties.


Effects of apomorphine upon local cerebral blood flow. The effects of apomorphine (0.1 mg/kg) upon local cerebral blood flow were examined in anaesthetized baboons. Proportionately similar increases in local cerebral blood flow were observed following apomorphine in the caudate nucleus (40%), cerebral cortex (38%) and cerebellar cortex (26%), despite the marked differences in the density of the dopaminergic innervation to these structures.


Visual cortex ablation and thresholds for successively presented stimuli in rhesus monkeys: II. Hue. Rhesus monkeys were trained to discriminate successively presented hues. The smallest difference they could reliably detect was determined before and after either inferotemporal ablation, or a lesion intended to remove as much as possible of prestriate area V4 (Zeki, 1973). As a group, the animals with lesions of V4 showed good but not perfect retention of their preoperative performance, and their thresholds were unaltered. The inferotemporal group showed no retention of the simplest successive task, red versus green, but after relearning their thresholds too were unaltered. It appears that animals without inferotemporal cortex can form precise internal representations of hues, and that the basis of the inferotemporal learning impairment may depend upon the nature of the stimuli to be discriminated.