Isolation of human antibodies to factor VIII. It has been claimed that human anti-VIII:C antibodies do not form stable complexes with factor VIII and this fact has hampered in the past the isolation of such antibodies. In this study the purification of human anti-VIII:C antibodies appearing in haemophiliac patients following replacment therapy has been achieved using two different systems. In a liquid phase system, purified human factor VIII was mixed with IgG from a haemophilic patient with a high titre antibody. Specific anti-VIII:C antibodies were recovered following filtration of the antigen-antibody complexes on Biogel A-5m, dissociation of complexes at pH 3.5 and final isolation by filtration on Sephadex G-200. In a solid phase system, the same IgG fraction was specifically bound to insolubilized human factor VIII. Purified anti-VIII:C antibodies were subsequently recovered by elution of antigen-antibody complexes with magnesium chloride. The results demonstrated that stable complexes from between anti-VIII:C antibodies and either the whole factor VIII molecule, or VIII:C dissociated by previous interaction with the antibodies. It is postulated that, in vivo, similar antigen-antibody complexes may form following replacement therapy in haemophilic patients with antibody.


Purification and characterization of an alpha-glucosidase from Saccharomyces carlsbergensis. alpha-Glucosidase (EC 3.2.1.20) was purified to homogeneity from logarithmically growing cells of Saccharomyces carlsbergensis. The purification involved the following steps: (a) ammonium sulfate fractionation; (b) Sephadex G-100 chromatography; (c) DEAE-cellulose chromatography; and (d) hydroxylapatite chromatography. This procedure gave a preparation judged to be greater than 98% pure by Na-DodSO4-polyacrylamide gel electrophoresis. The enzyme was shown to be a monomer of 63 000 daltons by gel filtration on Sephacryl S-200 under native conditions and by polyacrylamide gel electrophoresis under denaturing conditions. The Km values of the enzyme for the substrates maltose and p-nitrophenyl alpha-D-glucoside were found to be 1.66 X 10(-2) and 3.1 X 10(-4) M, respectively. The corresponding Vmax value for maltose was 44.8 X 10(-6) mol min(-1) mg(-1) and that for p-nitrophenyl alpha-D-glucoside was 134 X 10(-6) mol min-1 mg-1. The pH optimum for the purified enzyme was found to be between pH 6.7 and 6.8. The enzyme has an absolute anomeric specificity for alpha-glycosidic linkages and appears to recognize a glucosyl residue in alpha linkage on the nonreducing end of its substrate. For the strain used in this study, which carries the MAL 6 locus, only a single form of the enzyme was detected.


Proton correlation nuclear magnetic resonance study of anaerobic metabolism of Escherichia coli. Proton correlation nuclear magnetic resonance has been used to investigate anaerobic metabolism of glucose in Escherichia coli cells. The time course of the concentrations of six metabolites (ethanol, lactate, acetate, pyruvate, succinate, and formate) has been followed at the very early state of fermentation, and used to discuss dynamical aspects of the mixed-acid fermentation of glucose by E. coli.


Three hydroxylations incorporating molecular oxygen in the aerobic biosynthesis of ubiquinone in Escherichia coli. The biosynthetic origin of the oxygen atoms of ubiquinone 8 from aerobically grown Escherichia coli was studied by 18O labeling. An apparatus was developed which allowed the growth of cells under a defined atmosphere. Mass spectral analysis of ubiquinone 8 from cells grown under highly enriched 18O2 showed that three oxygen atoms of the quinone are derived from molecular oxygen. It was established that the molecular oxygen is incorporated into the two methoxyl groups (at C-5 and C-6) and one of the carbonyl positions of the ubiquinone molecule by demonstrating that only one of the incorporated oxygens will exchange with water under acidic conditions that specifically catalyze the exchange of carbonyl, but not methoxyl, oxygens. That the C-4 carbonyl oxygen is derived from molecular oxygen was shown by the incorporation of three atoms of 18O2 into ubiquinone 8 biosynthesized from added 4-hydroxybenzoic acid. Comparison of ubiquinone 8 and menaquinone 8 from E. coli grown under 18O2 confirmed that the labeled carbonyl oxygen of the [18O2]ubiquinone 8 is incorporated biosynthetically and not by chemical exchange in the cell. It is concluded that the three hydroxylation reactions involved in the pathway for the aerobic biosynthesis of ubiquinone are all catalyzed by monooxygenases. The implications of this study for the anaerobic biosynthesis of ubiquinone 8 in E coli are discussed.


Alternative hydroxylases for the aerobic and anaerobic biosynthesis of ubiquinone in Escherichia coli. The synthesis of ubiquinone under anaerobic conditions was examined in a variety of strains of Escherichia coli K12. All were shown to synthesize appreciable quantities of ubiquinone 8 when grown anaerobically on glycerol in the presence of fumarate. Under these conditions, ubiquinone 8 was in most cases the principal quinone formed, and levels in the range 50--70% of those obtained aerobically were observed. Studies with mutants blocked in the various reactions of the aerobic pathway for ubiquinone 8 synthesis established that under anaerobic conditions three alternative hydroxylation reactions not involving molecular oxygen are used to derive the C-4, -5, and -6 oxygens of ubiquinone 8. Thus, mutants blocked in either of the three hydroxylation reactions of the aerobic pathway (ubiB, ubiH, or ubiF) are each able to synthesize ubiquinone 8 anaerobically, whereas mutants lacking the octaprenyltransferase (ubiA), carboxy-lyase (ubiD), or methyltransferases (ubiE or ubiG) of the aerobic pathway remain blocked anaerobically. The demonstration that E. coli possesses a special mechanism for the anaerobic biosynthesis of ubiquinone suggests that this quinone may play an important role in anaerobic metabolism.


Inhibition of unsaturated fatty acid synthesis in escherichia coli by the antibiotic cerulenin. Low concentrations of cerulenin inhibit the growth of Escherichia coli by selectively blocking unsaturated fatty acid synthesis. This inhibition was relieved by unsaturated fatty acid supplements alone but not by saturated fatty acid supplements. The utilization of exogenous unsaturated fatty acids to sustain growth in the presence of cerulenin was confirmed by the analysis of bulk lipid composition. The effects of cerulenin on fatty acid synthesis were examined in vivo by pulse labeling with [14C]acetate and in vitro using [14C]malonyl-coenzyme A. In both cases, unsaturated fatty acid synthesis was inhibited by low concentrations of cerulenin with a stimulation of saturated fatty acid synthesis. Using mutant strains deficient in fatty acid synthesis, the effects of cerulenin on beta-ketoacyl-[acyl-carrier-protein] synthetases I and II were examined. Our results indicate that beta-ketoacyl-[acyl-carrier-protein] synthetase I is more sensitive to inhibition by cerulenin than beta-ketoacyl-[acyl-carrier-protein] synthetase II.


Characterization of the guanosine 5'-triphosphate 3'-diphosphate and guanosine 5'-diphosphate 3'-diphosphate degradation reaction catalyzed by a specific pyrophosphorylase from Escherichia coli. Guanosine 5'-triphosphate 3'-diphosphate (pppGpp) and guanosine 5'-diphosphate 3'-diphosphate (ppGpp) are specifically degraded by a manganese-dependent pyrophosphorylase present in spoT+ but not in spoT- strains of Escherichia coli, indicating that the enzyme is the spoT gene product. The enzyme catalyzes the release of pyrophosphate from the 3' position of ppGpp or pppGpp, yielding ppG and pppG, respectively; pppGpp could not be detected as an intermediate in the decay reaction. Degradation of (p)ppGpp is optimal in the presence of 200 to 300 mM potassium or sodium acetate, at a pH of 7.5 to 8 and a temperature of 37 degrees C.


Phosphate content of Escherichia coli alkaline phosphatase isozymes. The effect of phosphate and zinc on the separation of isozymes. Alkaline phosphatase from Escherichia coli was isolated as two major isoenzyme forms that were separated by DEAE-cellulose chromatography. Each form contained 2 equiv of endogenous phosphate. The endogenous phosphate, although difficult to remove, readily exchanges with phosphate. The forms also were separable by polyacrylamide gel electrophoresis. Apoenzyme prepared from native enzyme by the removal of zinc (and phosphate) also contains electrophoretically distinct enzyme forms which are indistinguishable from the native forms on gel electrophoresis. The isozymes were also found to have similar affinities for inorganic phosphate and susceptibilities to inactivation by EDTA. These results are not consistent with the notion that the formation or separation of isoenzyme forms is dependent upon different amounts of bound phosphate. They are consistent with the suggestion that a difference in amino acid composition is the basis for the occurrence and separation of these isoenzymes.


Photoaffinity labeling of the ribosomal peptidyl transferase site with synthetic puromycin analogues. A photoaffinity labeling puromycin analogue, Nepsilon-(2-nitro-4-azidophenyl)-L-lysinyl puromycin aminonucleoside (NAP-Lys-Pan), was synthesized and used for investigation of the peptidyl transferase center of 70S riobsomes. Visible light irradiation of NAP-Lys-Pan led to covalent linkage of the analogue with Escherichia coli ribosomes. In a subsequent step, poly(uridylic acid) was employed to direct Ac[14C]Phe-tRNA to the P sites of the photolabeled ribosomes. Transpeptidation of Ac[14C]phenylalanine to the bound NAP-Lys-Pan resulted in selective incorporation of radioactive label into the peptidyl transferase A site. Dissociation of the ribosomes into subunits, and digestion of the RNA components, indicated that the radioactive label was incorporated into a protein fraction of the 50S subunit.


Transcription of histone-covered T7 DNA by Escherichia coli RNA polymerase. Purified core histones (H2A, H2B, H3, and H4) and bacteriophage T7 DNA have been reconstituted to form a nucleoprotein complex, and the properties of this complex as a template for transcription by Escherichia coli RNA polymerase have been studied. At low ionic strength, RNA chain elongation rates are slow, and the chains produced even after long incubation are short. At higher salt concentrations, chain-elongation rates approach those on naked DNA. Since the salt concentrations used are not in themselves sufficient to dissociate the histones from the DNA, some mechanism must exist that permits passage of the polymerase through histone-covered regions.


Communication between white cells and the abnormalities of this in leukemia. The production of white blood cells is mediated by cellular communication. Proliferation and differentiation of granulocytic and monocytic progenitor and immature cells have been studied in detail and are regulated by stimulatory and inhibitory molecules produced and released from the progeny of the progenitor cells. The resultant interactions between cells and cell-derived molecules suggest operable positive and negative feed-back mechanisms during normal hematopoiesis, and what one sees in the end is the net result of these interactions. The complexities of cellular regulation are partially unravelled by physical separation of the different populations and biochemical analysis of the regulatory molecules. Subpopulations of granulocyte-monocyte progenitors (CFU-c) exist and evidence suggests that they vary in responsiveness to different molecules. Stimulatory molecules are themselves chemically and physically heterogenous and, until recently, believed to have similar biological actions, but this concept must be re-evaluated. Different molecules may activate different subsets of progenitor cells, and there is now a role for substances which enhance the stimulatory interactions. Many studies on normal and leukemic cell responsiveness to stimulation must be re-examined in light of this recent information. Several inhibitory substances operate during normal hematopoiesis. Mature granulocytes, progeny of CFU-c, appear to elaborate at least two inhibitory activities. One activity influences immature, recognizable granulocytes and the other indirectly reduces progenitor cell proliferation by decreasing the production and release of molecules which stimulate CFU-c. In addition, mononuclear phagocytes produce and release E-type prostaglandins in direct response to elevated levels of the stimulatory molecules and E-type prostaglandins counteract increased stimulatory levels by decreasing the sensitivity of CFU-c to stimulators. Much of our present level of sophistication derives from in vitro experimentation and it is apparent that we are only beginning to understand these inter-relationships and their relevance to the in vivo situation. However, these in vitro studies have shed light on the interactions occurring during leukemia. Leukemic cells retain the capacity to respond to normal regulators and must therefore be considered dependent rather than autonomous neoplasms. Abnormalities do exist in leukemic cell interactions: the progenitor cells themselves may be defective and leukemic cells may respond to molecules which normal cells do not. The degree of sensitivity to stimulators and inhibitors will have to be carefully investigated to determine if and what differences may exist between normal and leukemic cells. Normal mature granulocyte derived inhibitory activity is quantitatively deficient in leukemic cells but another inhibitory activity which appears to be specifically present in cells from patients with leukemia and some cases of myelodysplasia is present...


Partial characterization of protein kinase-catalyzed phosphorylation of low molecular weight proteins in purified preparations of pigeon heart sarcolemma and sarcoplasmic reticulum. Pigeon heart microsomes contain three minor size protein kinase substrates of minimal molecular weights of 22 000, 15 000, and 11500, as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When the microsomes were partially loaded with calcium oxalate and subjected to rate zonal and isopycnic centrifugations in sucrose density gradient columns, the 22 000 and the 15 000 dalton proteins settled in the heaviest fraction, which was composed mainly of vesicles of sarcoplasmic reticular membranes; the 11 500 dalton protein was concentrated in the lightest fractions, which consisted chiefly of vesicles of sarcolemmal origin. During incubation of the membrane fractions with Mg [gamma-32P]ATP significant amounts of 32P were incorporated into all these proteins. Incorporation of 32P into the 15 000 dalton protein was moderately and 32P incorporation into the 22 000 dalton protein was markedly enhanced in the presence of exogenous soluble cyclic AMP-dependent protein kinase and cyclic AMP. The phosphorylation of the three proteins was virtually unaffected by Ca2+ concentrations up to 0.1 mM and by ethyleneglycol-bis-(beta-aminoethyl-ether)-N,N'-tetraacetic acid in the absence of added Ca2+. Phosphorylation of the 22 000 and the 11 500 dalton proteins occurred mainly at serine residues. In the 15 000 dalton protein threonine residues were the main site of endogenous phosphorylation. Nearly equal amounts of [32P]-phosphate were incorporated into threonine and serine residues of this protein, when phosphorylation was supported by exogenous cyclic AMP-dependent protein kinase and cyclic AMP. The 15 000 dalton protein could be removed from its membrane attachment by extraction with an acidic chloroform/methanol mixture. This step opens the way for the purification of this membrane-bound protein kinase substrate.


Quinones as hydrogen carriers for a late step in anaerobic heme biosynthesis in Escherichia coli. A late step in anaerobic heme synthesis, the oxidation of protoprophyrinogen with fumarate as electron acceptor, was studied in extracts and particles of Escherichia coli mutants deficient in quinones or cytochromes. Mutants specifically deficient in menaquinone did not couple protoporphyrinogen oxidation to fumarate reduction, whereas mutants containing menaquinone but deficient in either ubiquinone or cytochromes exhibited this activity. These findings indicate that this coupled reaction is dependent upon menaquinone as hydrogen carrier but independent of ubiquinone and cytochromes. Other characteristics of this coupled reaction were also studied. The activity was located exclusively in the membrane fraction of cell-free extracts. Coproporphyrinogen III could not replace protoporphyrinogen as substrate. Methylene blue, triphenyl tetrazolium and nitrate, but not nitrite, could replace fumarate as anaerobic hydrogen acceptor. These findings have implications for the mechanism and regulation of microbial heme and chlorophyll synthesis and for the physiology of cytochrome synthesis in anaerobic microorganisms.


Plasma membrane vesicles isolated from epimastigote forms of Trypanosoma cruzi. Membrane vesicles can be obtained from epimastigote forms of Trypansoma cruzi by incubating cells with either cross-linking reagents or acid pH. Acetate, phtalate or citrate, at pH 4.0, but not at higher pH values, were able to induce plasma membrane vesiculation. Vesicles have been purified by sucrose density centrifugation and their membrane origin was demonstrated by the following criteria: (a) Vesicles are 5--10 times richer in protein-bound iodine when they are prepared from cells previously labeled with 131I by the lactoperoxidase catalyzed reaction. (b) Electron microscopy of vesiculating cells shows physical continuity between cell plasma membrane and vesicle membrane. (c) Antibodies prepared against purified vesicles are able to agglutinate epimastigote forms of T. cruzi with sera dilutions up to 1 : 256 to 1 : 512. (d) Freeze-fracture studies of the purified vesicles have shown images of faces P and E compatible with known images of the intact cell plasma membrane. Typical preparations of acetate vesicles present the following characteristics: total carbohydrate : protein=1.5--2.0; orcinol : protein-0.07 and absence of diphenylamine reaction. Vesicles contain 0.2--0.5% and 0.3--1.0% of the total homogenate protein and carbohydrate, respectively. The presence of 10 major protein bands and 30--50-fold enrichment of the four sugar-containing macromolecules present in epimastigote forms of T. cruzi have been demonstrated in these preparations.


Purification of an adenylyl cyclase-containing plasma membrane fraction from Trypanosoma cruzi. A fraction containing plasma membrane fragments has been purified from epimastigote forms of Trypanosoma cruzi. Cells were broken by sonic vibration under well defined conditions and membranes were isolated by differential centrifugation and equilibrium centrifugation in sucrose gradients. The co-purification (approximately 10-fold) of adenylyl cyclase and plasma membrane-bound radioactive iodine is highly suggestive of the localization of this enzyme in the plasma membrane of T. cruzi. Determination of succinate cytochrome c reductase and glucose-6-phosphatase activities, as well as of total amounts of DNA and RNA in the purified fraction, indicates a negligible contamination from other cellular organelles. The co-purification of acid phosphatase activity with bound labeled iodine and adenylyl cyclase was taken as circumstantial evidence that part of this enzyme also belongs to the plasma membrane of T. cruzi. Conventional electron miscroscopy and freeze-fracture images of this fraction are consistent with a highly enriched plasma membrane preparation.


Hydrolysis of low molecular weight isomaltosaccharides by a p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase from a thermophile, Bacillus thermoglucosidius KP 1006. A p-nitrophenyl-alpha-D-glucopyranoside-hydrolyzing alpha-glucosidase of a thermophile, Bacillus thermoglucosidius KP 1006, was purified to an electrophoretically-homogeneous state. Its molecular weight was estimated as 60 000 by gel electrophoresis. The molecular activity (ko) and the Km value at 60 degrees C and pH 6.8 for p-nitrophenyl-alpha-D-glucopyranoside were 233 s-1 and 0.24 mM, respectively. The enzyme cleft the non-reducing terminal alpha-1,6-glucosidic bonds of isomaltose, panose, isomaltotriose, isomaltotetraose, and isomaltopentaose. The ko values were 72.4, 194, 208, 233 and 167 s-1, and the Km values were 3.3, 9.5, 11, 13 and 21 mM, respectively. Each isomaltosaccharide was hydrolyzed to glucose by the cleavage of single glucose units from its nonreducing end. The present study suggests that the enzyme is an oligo-1,6-glucosidase (dextrin 6-alpha-glucanohydrolase, EC 3.2.1.10) and an exo-glucosidase.


Starch metabolism in Pseudomonas stutzeri. I. Studies on maltotetraose-forming amylase. The extracellular maltotetraose-forming amylase of Pseudomonas stutzeri was purified to homogeneity by a combination of affinity and hydroxyapatite chromatography. Sodium dodecyl sulfate-gel electrophoresis indicated that the oligomeric enzyme contains two different subunits with molecular weights of 48 000 and 58 000. Cross-linking studies using dimethyl suberimidate have demonstrated that the native enzyme consists of dimers. Seven isozymes of the amylase have been identified after polyacrylamide gel electrophoresis and amylose-digestion zymograms. The amylase of Ps. stutzeri is known to produce maltotetraose from linear and branched alpha-glucans by an exomechanism. The relatively high conversion rate of starch (75% hydrolysis), and the hydrolysis of cross-linked blue starch by this amylase indicate that the enzyme can cleave its substrates also by an endomechanism. Further strong evidence for an endomechanism was obtained from the action of the amylase on maltotetraose units which are located within the pullulan molecule. Dextran, pullulan, and maltotetraose are compeititve inhibitors. EDTA caused reversible inactivation. Amylase activity could be restored by addition of Ca2+. Heavy metals are inhibitory.


Hypertension self-control with a portable feedback unit or meditation-relaxation. Thirty borderline essential hypertensives were randomly assigned to a portable constant-cuff blood pressure feedback technique or meditation-relaxation. Each technique was taught in the laboratory, then practiced twice daily at home for four weeks. Subjects mailed daily records of their progress. Seven feedback and ten meditation-relaxation subjects completed the program. Both techniques produced significant systolic and diastolic reductions within practice sessions and diastolic reductions over weeks of training. Neither technique improved reductions nor reduced initial systolic pressure levels over the four weeks. Differences between biofeedback and meditation-relaxation in within-session pressure reductions were not significant.


Biofeedback-assisted relaxation training for primary dysmenorrhea: a case study. Primary dysmenorrhea is a familiar complaint to medical practitioners. Recently, behavior therapy has been shown to be an effective treatment for the symptoms of dysmenorrhea. The present case study offers biofeedback-assisted relaxation treatment as an effective alternative treatment. The Menstrual Symptom Questionnaire was used to classify dysmenorrhea as spasmodic or congestive. This classification provides homogeneous groups of patients. The patient in this study had an 18-year history of primary dysmenorrhea that was resistant to hormonal and analgesic treatment. After two months of baseline observation, she was given eight sessions of skin-temperature biofeedback and autogenic training. She reported significant reduction of pain and discomfort with the use of biofeedback-assisted relaxation. Desensitization using visual imagery, an important component of previous therapies, was not used. Further examination of the efficacy of biofeedback-assisted relaxation training for the treatment of both congestive and spasmodic dysmenorrhea is suggested.


Relative rates of repair of single-strand breaks and postirradiation DNA degradation in normal and induced cells of Escherichia coli. Labeled DNA from irradiated Excherichia coli cells has been studied on an alkaline sucrose gradient without acid precipitation of the DNA. This enables the observation of both DNA repair and DNA degradation. The use of a predose of ultraviolet light (UV) causes induction of an inhibitor of postirradiation DNA degradation in lex+ strains. The effect of this induction on both the repair of single-strand breaks and DNA degradation has been followed in strains WU3610 (uvr+) and WU3610-89 (uvr-). The repair process is more rapid than the degradation, and when degradation is inhibited more repair is apparent. Cells that are lex- (Bs-1 and AB2474) cannot be induced for inhibition of degradation. Nevertheless, by observation at short times repair can be seen clearly. This repaired DNA is degraded, suggesting that the signal for DNA degradation is not a single-strand break.


Rate-determining processes in the transport of Pr3+ ions by the ionophore A23187 across phospholipid vesicular membranes. A 1H-MR and theoretical study. Rate constants and activation parameters have been determined for the transport of Pr3+ ions by the ionophore A23187 across dipalmitoyl phosphatidylcholine vesicular membranes. The novel method described depends on the measurement of changes in chemical shift of the 1H-NMR choline head-group signals as Pr3+ is transported from outside to inside the vesicles. The determined rates are directly proportional to A23187 concentration, suggesting that the rate-determining step involves the species [Pr(A23187)](2+). A theoretical analysis of the initial stages of Pr3+ transport leads to the conclusion that diffusion over the image potential barrier is the rate-determing step. Calculation of the form and height of this barrier for the non-equilibrium state gives results which agree well with the experimental activation energy and also correctly predict a two-fold reduction in rate of transport when 7 mol % decane is present in the bilayer.


Procedure for the simultaneous large-scale isolation of pullulanase and 1,4-alpha-glucan phosphorylase from Klebsiella pneumoniae involving liquid-liquid separations. A procedure for the simultaneous large-scale isolation of pullulanase and 1,4-alpha-glucan phosphorylase from Klebsiella pneumoniae is described. The pullulanase is solubilized from the cell wall by cholate treatment; cells and cell debris are removed by partition in a poly(ethylene glycol) (PEG)-dextran two-phase system and from the upper (PEG) phase of this system the pullulanase is isolated by ultrafiltration and precipitation with N-cetyl,N-,N-,N-trimethyl ammonium bromide to a purity of about 80% with a yield of 70%. The preparations are free of alpha-amylase activity. The cell containing dextran-rich phase is passed through a Manton-Gaulin homogenizer. Then the phosphorylase is separated from the cell debris by partition in a second PEG-dextran system. From the top phase of this system the phosphorylase is isolated by distribution in a PEG-salt two-phase system followed by batch adsorption on carboxymethyl-Sephadex in a yield of 55%, a purity of around 90%, and nearly free of glycosyltransferase activity. All steps in the isolation of the two enzymes can be performed easily in a large scale.


[Effect of leukocyte pyrogen on the phagocytic properties of macrophages in tissue culture]. The effect of rabbit leukocytic pyrogen (LP) on the phagocytic activity of the peritoneal macrophages of albino mice against shigellae was investigated. The dose-effect dependence was revealed: high LP doses depressed the phagocytosis, and low ones were insufficiently effective; addition of average LP doses against the kanamycin background stimulated both the absorption phase and that of shigella digestion. Phagocytosis stimulation with LP was also accompanied by an increase of the acid phosphatase lysosomal enzyme activity in macrophages; the RNA content was not changed.


Hodgkin's disease and anergy.

Hodgkin's disease and anergy. The clinical, pathologic, and immunologic features unique to Hodgkin's disease can be explained by the following hypothesis. A viral transformation of a lymph node cell leads to proliferation of tumor cells and the generation of an immune response consisting of lymphokine production, B cell activation and concomitant suppression of further T cell activation, but ineffective cellular cytotoxicity against the tumor cells. The result of this interaction would be chronic infiltration around the transformed cells, increased immunoglobulin synthesis, and anergy. Failure to destroy the target cells would result in chronicity of these features and progressive disease.


Effects of sulphinpyrazone and aspirin on prostaglandin I2 (prostacyclin) synthesis by endothelial cells. Synthesis of prostaglandin I2, (PGI2, prostacyclin) by vascular endothelium (assayed by the ability of cultured endothelial cells to inhibit platelet aggregation) was inhibited by aspirin. At 100 mumol/l aspirin completely blocked measurable PGI2 production, but endothelial cells had substantially recovered their ability to synthesize PGI2 24 h after removal of the drug. In contrast, the effect of 1 mmol/l aspirin was still evident 24 h after drug withdrawal. Sulphinpyrazone also inhibited PGI2 synthesis, but was about 100 fold less potent than aspirin, and the effect of the drug was lost within 24 h of its addition, even when endothelial cells were left in contact with the drug during this period.


Comparison of the actions of octopamine and catecholamines on single neurones of the rat cerebral cortex. 1. The technique of microelectrophoresis was used to compare the actions of octopamine, noradrenaline and dopamine on single cortical neurones of the rat. 2. Octopamine both excited and depressed neurones of the cortex. Frequently cells depressed by noradrenaline were excited by octopamine; occasionally the converse was true. The time courses of action of the two amines also differed. Dopamine-elicited excitations were observed, but also were not correlated with octopamine-elicited effects. 3. When octopamine and noradrenaline both caused depressant effects, octopamine frequently was of less apparent potency than noradrenaline. When these amines were excitatory, octopamine appeared at least as and sometimes more potent than noradrenaline. 4. Octopamine was only weakly effective on cortical neurones identified by antidromic stimulation of the pyramidal tract, or synaptically excited by stimulation of the ventrobasal thalamus. 5. alpha-Flupenthixol and propranolol were without effect on octopamine-elicited changes in firing rate at doses which were effective in blocking the actions of dopamine and noradrenaline respectively. Metoclopramide did not block the actions of any of the three agonists, but had strong effects of its own. 6. The results suggest that receptors sensitive to octopamine, and which appear to be pharmacologically distinct from those previously categorized as noradrenaline and dopamine receptors, may exist on central neurones of the rat.


A controlled study to evaluate directed masturbation in the management of primary orgasmic failure in women. This paper presents the results of a prospective controlled study evaluating a programme of directed masturbation against a combined sensate focus and supportive psychotherapeutic approach in the management of female primary orgasmic failure. Of the 20 patients who followed the masturbation programme 90 per cent gained orgasmic capacity compared with 53 per cent of 15 patients who were treated conventionally. Eighty-five per cent of the patients treated by the masturbation programme and 47 per cent of the control group of patients became coitally orgasmic on at least 75 per cent of coital occasions. The difference is statistically significant at the 5 per cent level. The results suggest that directed masturbation is an effective adjunct in the management of primary female orgasmic failure.


Mumps prophylaxis in the light of a new test for antibody. A radial haemolysis test was used to investigate immunity to mumps. Antibody was found in 92 (42%) out of 220 children aged up to 5 years, 124 (78%) out of 159 children aged 6--10 years, 192 (86%) out of 222 children aged 11 years, 138 (92%) out of 150 children aged 15 years, and 280 (95%) out of 296 women attending an antenatal clinic. A group of 307 cadets aged 16--18 years were also tested and interviewed: 133 (95%) out of 140 who said that they had had mumps and 108 (87%) out of 124 who said that they had not had mumps were found to have antibody. The results suggest that tests for immunity to mumps by radial haemolysis would permit more rational use of mumps-specific immunoglobulin and attenuated mumps vaccine.


Physiotherapy and intermittent positive-pressure ventilation of chronic bronchitis. Patients aged over 45 admitted to hospital with exacerbations of chronic bronchitis alone or in association with cor pulmonale, pneumonia, or respiratory failure were placed in one of three groups (men with hypoxia, men without hypoxia, and women). Patients within these groups were then randomly allocated to receive either standard drug treatment alone or standard drug treatment plus intermittent positive-pressure ventilation (IPPV). No significant differences occurred between the controls and patients receiving physiotherapy and IPPV in any group. We conclude that when a patient with chronic bronchitis and respiratory failure is deteriorating emphasis should be on correct diagnosis, fluid electrolyte balance, and nutrition together with oxygen treatment when necessary, rather than on additional physiotherapy.


Use of neurosurgery for psychological disorder in British Isles during 1974--6. All 44 neurosurgical units in the British Isles replied to a postal questionnaire asking about their use of neurosurgery during 1974--6 for functional mental illness. A total of 431 operations was reported, representing a yearly rate of 3.4 operations per million population aged over 15. The numbers of operations declined from 158 in 1974 to 119 in 1976. Four units did two-thirds of the operations. Stereotactic methods for locating the site for the lesion were used in two-thirds of procedures. Mood disorders, anxiety states, and obsessive-compulsive neurosis were the conditions most commonly treated.


Synthesis of dopamine and octopamine in the crustacean stomatogastric nervous system. The spiny lobster stomatogastric nervous system synthesizes dopamine and octopamine in vitro from exogenous [3H]tyrosine. Each amine accumulates with a specific distribution among 9 separately analyzed regions within the system. Synthesis of other catecholamines was not observed. [3H]Dopamine is found in nerves, ganglia, and identified commissural ganglion cell bodies in which catecholamine histofluorescence has been demonstrated. The biosynthetic and histochemical data together indicate that dopaminergic cells send axons from the commissural ganglia to the stomatogastric ganglion neuropil along the same pathway followed by fibers that activate the pylroic motor network. The results support the hypothesis that dopamine mediates activation of the pyloric system in vivo, as observed in vitro. [3H]Octopamine accumulates primarily in the commissural and stomatogastric ganglia, where it may modulate neuronal activity, but octopaminergic cells and release sites within the stomatogastric system have not been identified.


Protection of the brain from hypoxia: a review. A functional classification of hypoxia of the brain has been presented and some of its significant aspects have been discussed. Mechanisms of protection from hypoxia of the brain were reviewed under the headings of prevention, hyperventilation, hypothermia and protection by barbiturates. In prevention of hypoxia of the brain, avoidance of factors producing a fall in cerebral perfusing pressure was emphasized. Hyperventilation is not advised unless one can readily measure regional cerebral blood flow. In the operating room, normocarbia or slight hypocarbia is recommended. Animal studies indicate a protective role of barbiturates in ischaemic hypoxia of the brain. However, it should be emphasized that, at present, hypothermia is the only established means of protection against hypoxia of the brain in man, when it is induced prior to the hypoxic insult. The evidence for protection by barbiturates has been found only in experimental animals. If one can extrapolate the results of studies in animals to man, then potential benefits would be expected in clinical stroke, cardiac arrest, in operations on the carotid artery and in head injury.


Comparison of three methods for identifying nonfermenting gram-negative rods. Seventy-six strains of nonfermenting gram-negative rods were tested on the Analytab Products, Inc. (API) system and on conventional media. In addition, 51 strains were tested on the Oxi-Ferm system. When the identification results were compared, the API agreed with the conventional system on 41% of the isolates and Oxi-Ferm agreed with the conventional system on 72% of the isolates. API had the greatest difficulty in identifying Pseudomonas aeruginosa. API and Oxi-Ferm both had difficulty identifying P. cepacia. Oxi-Ferm had more individual discrepant biochemical reactions than did API when compared to the conventional media, but still maintained a higher percentage agreement with the conventional system.


Simultaneous production of two types of beta-lactamase in Escherichia coli and Providencia stuartii. The production of beta-lactamase has been studied in two strains isolated from clinical samples: Providencia stuartii MULB 501 and Escherichia coli MULB 130. These strains were selected for their high resistance level to penicillins and cephalosporins. Determination and identification of beta-lactamase activity were achieved by combining several up-to-date methods including (i) neutralization by anti-beta-lactamase sera, (ii) purification by affinity chromatography on cephalosporin C linked Sepharose 4B, (iii) determination of substrate specificity and kinetic values (Km and Vmax) by a computerized microacidimetric method, and (iv) isoelectric focusing. The results clearly demonstrate that in these two strains there is a simultaneous production of different beta-lactamases: the first one is similar to the TEM penicillinase and the second one shares a typical cephalosporinase profile. This double beta-lactamase production is a relatively rare phenomenon.


Inhibitors of DNA synthesis cause excessive DNA synthesis in dnaA mutants of Escherichia coli K12. The relative rate of net DNA synthesis was stimulated when cells of dnaA mutants of Escherichia coli K12 were grown in the presence of low concentrations of DNA synthesis inhibitors. This led to a supernormal DNA/cell mass ratio. The excessive DNA was similar to the normal chromosomal DNA in size and stability in vivo. However, the cells did not divide but turned into long filaments. Excessive DNA synthesis in the presence of inhibitors of DNA synthesis was observed in the cultures of two independent dnaA mutants of E. coli, but dnaB and dnaC mutants behaved like the wild type in this respect.


A new technique for the preparation of permanent reference cultures of fungi. Fungal cultures were grown by a new microslide method and by the traditional slide-culture method. Sporulation results were similar by both methods. The microslide method has the advantages of increased safety to the user and ease of storage.


Temperature-induced differences in growth rate and fatty acid composition in two strains of Escherichia coli 15T-. The saturated/unsaturated fatty acid ratio of Escherichia coli 15T- decreases almost threefold as growth temperature decreases from 43 to 27 degrees C, wheras the ratio of a fast-growing mutant derived from 15T- changes only half as much. Strain 15T- experiences a 2.4-fold change in doubling time across this temperature range, but doubling time in the mutant changes 3.3-fold.


Chemotherapy of advanced measurable colon and rectal carcinoma with oral 5-fluorouracil, alone or in combination with cyclophosphamide or 6-thioguanine, with intravenous 5-fluorouracil or beta-2'-deoxythioguanosine or with oral 3(4-methyl-cyclohexyl)-1(2-chlorethyl)-1-nitrosourea: a Phase II-III study of the Eastern Cooperative Oncology Group (EST 4273). In a randomized multi-institutional trial of the Eastern Cooperative Oncology Group, 316 patients with advanced measurable colorectal adenocarcinoma were treated with a weekly schedule of 5-fluorouracil given orally and intravenously with oral-5-fluorouracil in combination with cyclophosphamide or 6-thioguanine, or with oral Methyl CCNU administered once every eight weeks. On failure or progression, 133 protocol patients crossed-over to a secondary therapy, while 116 other patients previously treated with 5-fluorouracil off protocol were randomized to treatment with Methyl CCNU or B-2'-deoxythioguanosine. Response rates among patients who had received no prior chemotherapy were 18% to oral 5-FU, 15% to intravenous 5-FU and to MeCCNU, 12% to 5-FU and 6-thioguanine and 5% to cyclophosphamide and 5-FU, with little activity (3% response rate) in crossover or previously treated patients. Treatment with 5-FU, particularly oral 5-FU was associated with the least drug-related toxicity. Hematologic toxicity was greatest with Methyl CCNU, but was no more frequent in previously treated than in untreated patients. A tendency toward cumulative bone marrow depression was noted. 5-FU was effective only in ambulatory patients, whereas responses among non-ambulatory patients were seen only in the group treated with Methyl-CCNU.


Cyclophosphamide (NSC 26271) versus the combination of adriamycin (NSC 123127), 5-fluorouracil (NSC 19893), and cyclophosphamide in the treatment of metastatic prostatic cancer: a randomized trial. Twenty-seven patients with a diagnosis of metastatic adenocarcinoma of the prostate were treated in a randomized, prospective trial with either Cyclophosphamide or a combination of Adriamycin, 5-Fluorouracil, and Cyclophosphamide. Doses were either Cyclophosphamide alone (800-1200 mg/m2 iv q 3 weeks) or Cyclophosphamide (150-200 mg/m2 po Day 3-6) plus 5-FU (400-500 mg/m2 iv Day 1, 8) plus Adriamycin (30-50 mg/m2 iv Day 1) given as a 4 week treatment cycle. Patients with compromised bone marrow reserve initially received the lower dose level. Objectively stable disease as defined by a modification of the National Prostatic Cancer Project criteria was seen in 53% of the 15 Cyclophosphamide treated patients and in 50% of the 12 combination treated patients. Survival was not significantly different in the two arms. However, the survival of patients responding to Cyclophosphamide was significantly longer than that of patients responding to the combination (median 18.6 months versus 8.1 months, p less than 0.05). Gastrointestinal and hematologic toxicity was moderate with both regimens. Therefore, in the present study, Cyclophosphamide alone was as effective as the combination of Cyclophosphamide, 5-FU and Adriamycin for patients with disseminated prostatic carcinoma. The moderate hematologic toxicity noted with both regimens suggests further evaluation of drug combinations utilizing higher dosages of active agents in this disease.


Radiation therapy in lymphomatoid granulomatosis. Two female patients with lymphomatoid granulomatosis have received radiation therapy for local, progressive, symptomatic lesions. One patient was a 56-year-old woman with concomitant brain and pulmonary lesions which progressed on cyclophosphamide and prednisone therapy. The second patient was a 62-year-old woman with a supraorbital mass which appeared and progressed on prednisone therapy. Both patients exhibited rapid response to radiation therapy, in terms of relief of symptoms and objective evidence of subsidence of local disease. A discussion of the nature of lymphomatoid granulomatosis and its treatment is presented with a review of the literature. One hundred sixty-five cases have now been described including the two in this study, and in only one previous case was radiation therapy utilized to treat local disease, prior to the two cases described herein.


Osteosarcoma associated with Thorotrast administration: report of two cases and literature review. Two cases of osteosarcoma are described arising in patients who had undergone angiography with Thorotrast during childhood. In both patients, there was radiographic and pathologic evidence of radioactive thorium dioxide deposition in bone as well as throughout the reticuloendothelial system. Thorotrast deposits were demonstrated in the immediate vicinity of the primary tumors by both histology and autoradiography. Previous cases of osteosarcoma associated with Thorotrast administration from the literature are cited, and possible causal relationships are discussed between thorium retention in bone and neoplastic transformation by chronic radiation.


Primary malignant melanoma of the vagina: study of four autopsy cases with ultrastructural findings. Four cases of primary malignant melanoma of the vagina in women aged 23, 44, 51 and 65 years are presented. In these 4 cases, thorough clinical and postmortem examinations ruled out the possibility of a primary melanoma elsewhere. The primary tumors showed exophytic growth with superficial ulceration. Three of the melanomas arose from the middle third of the vagina and one from the upper third. Melanin was visible in sections stained with hematoxylin and eosin in 3 of the tumors. In the other one, the first biopsy failed to reveal melanin. However, the second biopsy performed following irradiation showed abundant melanin pigment. Electron microscopic examination of 3 tumors revealed premelanosomes and melanosomes in the tumor cells, thus confirming the diagnosis. Two neoplasms showed atypical histologic features, and only the presence of melanin enabled us to make diagnosis of malignant melanoma. One melanoma was associated with an adjacent widespread intraepithelial component of superficial spreading type indicating its probable mode of origin. All 4 patients died of widespread metastases within 13 months after initial treatment. These 4 cases, in which clinical diagnosis was confirmed by thorough autopsy, strongly indicate that malignant melanoma can arise directly from the vagina.


Primary carcinoid tumor of the testis: case report, ultrastructure and review of the literature. A case of primary pure carcinoid tumor of the testis which occurred in a 71-year-old male is reported. The patient was treated by radical orchiectomy and remains well and symptom free 10 months after operation. Histologically as well as ultrastructurally the tumor showed typical appearances of carcinoid tumor of midgut derivation. 23 cases of carcinoid tumors of the testis were discovered in the literature. Of these 17 were primary testicular carcinoids, and 6 were metastatic to the testis. Of the 17 cases of primary carcinoid tumors, 14 were pure carcinoids and only 3 were associated with teratoma. None of the primary testicular carcinoids were associated with metastases and the prognosis after orchiectomy was excellent, thus indicating that no further therapy is necessary. The prognosis of patients with carcinoid metastatic to the testis is poor. In view of this it is very important to determine whether the tumor is primary or metastatic.


Meningeal mesenchymal chondrosarcoma: report of 8 cases with review of the literature. This paper reviews 8 personally examined cases of primary meningeal mesenchymal chondrosarcoma and 4 similar cases previously reported by others. The clinicopathologic features of these extraosseous intracranial and intraspinal examples are similar to those of other extraskeletal mesenchymal chondrosarcomas. The tumor occurred most often in the second and third decades, showed a moderate tendency to local recurrence (5 of 12 cases) and occasionally metastasized to the lungs (1 case). Both intracranial and intraspinal tumors occurred with equal frequency, but the former, probably due to the later onset of symptoms, had the worse prognosis. Microscopically, they are composed of primitive undifferentiated mesenchymal cells and frequently well-defined islands of hyaline cartilage. There is an apparent correlation between the frequency of mitotic figures and the likelihood of recurrence and metastasis. Electron microscopic study of one example revealed morphologic features similar to those previously described by others and supports the conclusion that the neoplastic cells represent primitive precartilaginous mesenchyme displaying focal cartilaginous differentiation.


Soft tissue metastasis of a chemodectoma: a case report and review of the literature. A case of a single soft tissue metastases (abdominal wall) occurring 15 years after removal of chemodectoma of the right carotid bifurcation is described. The tumor had apparently been completely removed with excision of portions of the common, external and internal carotid arteries up to the base of the skull. Seven years later osteolytic and osteoblastic lesions were noted in the cervical and thoracic vertebrae which were radiated with 3500 rad. The 19 previously reported cases of chemodectomas with distant metastases are reviewed.


Tracheal carcinoids.

Tracheal carcinoids. Three cases of carcinoid tumor arising in the trachea are reported and contrasted with carcinoids arising in bronchi and carcinoids in general. Only eleven other documented examples of tracheal carcinoids are found in the English literature. The true prevalence of carcinoids primary in the trachea cannot be accurately determined from the literature because of imprecise nomenclature or because of the failure to distinguish this tumor from carcinoids primary in the bronchus. Presenting symptoms are hemoptysis, dyspnea and wheezing, often persisting for many years before the correct diagnosis is made. The treatment of choice is surgical resection of the involved segment of trachea and primary reconstruction. The prognosis is generally good. The tumor metastasized in one of our three cases and in none of the eleven cases in the English literature.


Mutagenicity and recombinogenicity of daunomycin in Saccharomyces cerevisiae. The antineoplastic antibiotic daunomycin was tested for mutagenic and recombinogenic effects in haploid and diploid strains of Saccharomyces cerevisiae. This antibiotic failed to induce mutations to lysine and histidine independence in the haploid strain, XV 185-14C, carrying the auxotrophic markers lysl-1 and hisl-7. On the contrary, daunomycin did induce aberrant colonies, including reciprocal products of mitotic crossing over in the diploid strain D7. The results suggest that while daunomycin may be recombinogenic, its mutagenic effects, if any, are either highly specific or negligible. The significance of these results in evaluating the mutagenic/carcinogenic potential of antitumor agents is discussed.


Randomized trial of levamisole in patients with squamous cancer of the head and neck: a preliminary report. This is a preliminary report of a randomized double-blind study comparing levamisole with placebo as surgical adjuvant treatment of patients with squamous cancer of the head and neck. The study groups were comparable according to the prognostic factors of type (primary or recurrent), site (oral cavity, pharynx, or larynx), and stage of disease and were also similar in pretreatment immune function and in radiation exposure. Of 65 evaluable patients, 31 received levamisole and 34 received placebo. Although there was no difference in the distribution of the time to recurrence of the overall treatment group, there was a difference of borderline significance in favor of levamisole in the primary disease patients (P less than 0.06). Further analysis of subgroups in the primary disease category showed decreased recurrence rates in patients receiving levamisole who had cancer of the oral cavity (P less than 0.01) or who had stage II disease (P less than 0.02). Considering site and stage simultaneously, the only significant difference was in stage II patients with oral cancer. The above results, although encouraging, must be viewed with caution due to small sample sizes and the relatively short followup of most patients.


Final results of a multicenter placebo-controlled levamisole study of resectable lung cancer. Two hundred and eleven patients with resectable lung cancer have been treated in a double-blind clinical investigation with either levamisole (50 mg three times a day) or a placebo given in 3-day courses starting 3 days before the operation and repeated every 2 weeks. The study was terminated after all patients had been in the trial for 2 years. The dose of levamisole used in this study appears to be sufficient only for patients weighing less than or equal to 70 kg and, therefore, future studies should make use of an individually adapted dose. Such a treatment is expected to prolong both the disease-free interval and the survival time without causing major side effects in a significant number of patients. This result is achieved primarily by an inhibition of hematogenous dissemination and occurs especially in those patients who had more advanced tumors at the time of the resection.


Effect of levamisole as a surgical adjuvant therapy for malignant melanoma. The preliminary results of a trial in progress are reported. Two hundred patients have been enrolled in a randomized, double-blind, placebo-controlled trial of levamisole in malignant melanoma. A median followup of 20 months has been reached. Endpoints of the study include disease-free interval, interval free of visceral disease, and survival. Overall, there is a trend in favor of the levamisole group with regard to all three endpoints, but the difference is not significant. Significant differences between the control and levamisole groups are found at some, but not all, time intervals when the analysis includes only patients with stage II disease (positive lymph nodes at the initiation of the study). Although these preliminary results are encouraging, at the present time the overall curve for patients with stage II disease is not statistically significantly different when analyzed by the method of Mantel.


Preliminary report of the use of levamisole in the treatment of bladder cancer. Sixty-two patients with transitional cell carcinoma have been admitted to a double-blind randomized control study with levamisole as an immune adjuvant, in addition to standard therapy for noninvasive and invasive bladder cancer. Levamisole has been shown to be easily administered and is well-tolerated, especially when compared to other immune adjuvants such as bacillus Calmette-Guerin or Corynebacterium parvum. To date, there is no significant difference in the disease-free interval in the levamisole-treated group compared to the placebo group. Initial dinitrochlorobenzene (DNCB) reactivity may be an important prognostic indicator with regard to tumor recurrence. Tumor recurrence seems to be rare in those patients who are initially DNCB-positive. Total monocyte count, T lymphocyte, FC receptor cells, and PHA response showed no improvement with levamisole therapy. Monocyte chemotaxis remains the only immune function study to improve with levamisole, but the clinical significance of this test is yet to be explained.


Correlations of clinical and laboratory effects of treatment with levamisole in autoimmune disease. Studies in which levamisole has been used with success in the treatment of rheumatoid arthritis and other autoimmune diseases may provide information useful in cancer treatment with regard to optimal dosage regimens and methods for monitoring laboratory effects. Using the response of peripheral blood lymphocytes to phytohemagglutinin mitogen measured by the uptake of tritiated thymidine in 3-day culture, patients with rheumatoid arthritis were found to have a significantly deficient response compared to normal subjects. Levamisole treatment reversed this deficiency and normalized lymphocyte mitogen responses. These studies showed the benefit of a dose of 150 mg of levamisole compared to lower doses and also showed equal maintenance of immunocompetency with a dose of 150 mg/week compared to daily levamisole. In addition, compared to poor clinical responders, good clinical responders were shown to have a greater enhancement in mitogen response, both during the treatment period and when levamisole was added in vitro to lymphocyte culture prior to drug treatment. It was concluded that laboratory studies of lymphocyte mitogen response may contribute towards better management of patients receiving levamisole therapy.


Adverse reactions to levamisole.

Adverse reactions to levamisole. The adverse reactions mentioned in 267 reports on levamisole are analyzed. Significant adverse reactions are agranulocytosis, skin rash, and febrile illness. They occur in a small subpopulation of patients only, predominantly those with rheumatoid arthritis. Other side effects rarely necessitate discontinuation of treatment. Levamisole did not appear to cause renal or liver toxicity. By means of a questionnaire, cases of blood dyscrasia were analyzed in detail. Agranulocytosis (less than 20% neutrophils) occurs most frequently in patients with rheumatic diseases, in women, and in HLA B27 genotypes. It is spontaneously reversible when treatment is discontinued. The bone marrow is not damaged. Thrombocytopenia has a course similar to that of agranulocytosis but it is very rare. Leukopenia (less than 3000 cells/mm3 but greater than 20% neutrophils) seems to differ from agranulocytosis in a number of ways and is not an indication to discontinue treatment. It is concluded that, if used under the close supervision of a physician, for those clinical situations in which levamisole has proven efficacy, the potential benefit to the patient outweighs the hazards and justifies its use.


Future direction of synthetic polyanions (pyran copolymer). Pyran copolymer (maleic anhydride-divinyl ether) has consistently reproducible molecular weight-related biologic effects associated with toxic, immunologic antitumor, and antiviral effects. Fortunately, the antitumor action occurs with the least toxic lower molecular weight fraction. Immunoadjuvant effects with this fraction would be critical to its development. Studies of polymers should include evaluation of effects on splenomegaly, splenic esterase changes, lipolysis, reverse transcriptase, nucleases, calcium flux, cyclic nucleotides, and complement and clotting elements.


Postinfarction exercise capacity after lidoflazine treatment or physical training. 44 male postinfarction volunteers were divided into 4 groups and submitted to performance tests. Groups I and II consisted of 24 patients, 12 of whom followed a 2-month physical training program, while the other 12 served as controls. Groups III and IV (10 patients each) were included in a 10-month double-blind crossover study with lidoflazine 240 mg/day vs. placebo. The cardiovascular adaptation of the patients treated for 5 months with lidoflazine had two features in common with that observed after 2 months of physical training, namely an increase in maximal exercise capacity and, during submaximal exercise tests, a decrease in heart rate compensated for by an increase in stroke volume. In contrast to physical training, treatment with lidoflazine did not improve the peripheral oxygen consumption by the muscles.


Prolapsing right atrial myxoma.

Prolapsing right atrial myxoma. Echocardiography was of great value in establishing the correct diagnosis in a patient with symptoms of congestive heart failure due to a prolapsing right atrial myxoma. The movement of the tumor into the right ventricle through the atrioventricular orifice during diastole was identified by the echocardiogram and confirmed by angiography and direct observations during surgery. The indicator dilution curve also showed abnormal hemodynamic features simulating tricuspid incompetence, in relation to the movement of the tumor.


Immunohistochemical localisation of ecdysteroids in the follicular epithelium of locust oocytes. Frozen sections of growing terminal follicles of the locust ovary were incubated with an ecdysteroid-specific rabbit antibody and the bound antibody visualised by the use of FITC-labelled goat-anti-rabbit antiserum. A bright fluorescence was seen in the cytoplasm of the follicle cells in terminal follicles with a length between 4.0 and 6.0 mm with a maximum intensity at 5.5 mm, indicating the presence of ecdysteroids in these cells in this particular developmental stage.


Immunocytochemical localisation of prolactin and growth hormone in the ovine pituitary. A morphological and quantitative study. Using the peroxidase-antiperoxidase immunocytochemical staining technique, prolactin and growth hormone cells have been identified and described in the ovine pituitary. The image analysing computer, Quantimet 720, was used to assess accurately the size range of the secretory granules in these cell types. The area size distributions of the prolactin and growth hormone granules are similar. An increased proportion of larger granules was observed in the prolactin cells post-partum. Serial sections stained alternately for prolactin or growth hormone confirmed that the cells contain either prolactin or growth hormone but not both.


Light- and electron microscopic localization of vasopressin or a vasopressin-like substance in the neurons of the rat suprachiasmatic nucleus. In the suprachiasmatic nucleus of the rat light microscopic immunostaining for vasopressin reveals a distribution pattern of the immunoreactive material different from that known for the supraoptic nucleus. Among non-stained neurons positive-reacting perikarya display a cap- or tip-like labeling. The area of the suprachiasmatic nucleus is marked by delicate vasopressin-positive fibers. At the ultrastructural level the reaction product, after incubation with anti-vasopressin, is localized in small elementary granules unevenly distributed over the cytoplasm. Groups of axons containing specifically labeled granules contact non-reacting fibers.


Immunoreactive neurotensin and somatostatin in the chicken thymus. A chemical and histochemical study. Two distinct populations of endocrine cells in the chicken thymus display neurotensin and somatostatin immunoreactivity, respectively. Both cell types are few in number at hatching but proliferate rapidly during the first week. The neurotensin cells are 'Grimelius-positive' and 'Hellerström-Hellman-negative'. The somatostatin cells are 'Grimelius-negative' and 'Hellerström-Hellman-positive'. Both cell populations are non-argentaffin. The somatostatin-like material extracted from chicken thymus behaves immunochemically and chromatographically similar to synthetic ovine somatostatin, while the neurotensin-like material, from the thymus as well as from the gut, differs from synthetic bovine neurotensin in that it appears larger in size and more basic.


Cold-induced insulin release in vitro: evidence for exocytosis. Exposure of isolated pancreatic islets (mouse or rat) to low temperature (2 degrees C) evoked a threefold increase in insulin release irrespective of the glucose concentration in the incobation medium. Cold-induced release was transient and rewarming to 37 degrees C restored the sensitivity of B-cells to gluocose stimulation. In islets cooled to 2 degrees C, exocytotic profiles could easily be detected both by thin-section and freeze-fracture electron microscopy. As revealed by the freeze-fracture technique, the number of exocytotic profiles per membrane area was increased three- to fourfold as compared to islet cells incubated at 20 degrees C. This was paralleled by intracellular fusion of secretory vesicles. Cold-induced insulin release was not affected by theophylline, cytochalasin B, omission of extracellular Ca++ or D600. Replacement of extracellular Na+ with choline or sucrose suppressed the increase in insulin release and in frequency of exocytotic profiles recorded after exposure to 2 degrees C. It is suggested that a redistribution of Ca++ from intracellular stores, possibly mediated by an increase in intracellular Na+ triggers exocytosis of insulin granules upon exposure to cold.


Use of immunocytochemical staining of somatostatin for correlative light and electron microscopic investigation of D cells in the pancreatic islet of Xiphophorus helleri H. (Teleostei). Somatostatin-containing cells have been demonstrated by immunocytochemistry in semithin sections of the pancreatic islet of the teleost fish, Xiphophorus helleri. These cells were shown by correlative light and electron microscopy to be identical with D cells previously defined in this species by the silver impregnation method of Hellman and Hellerström.


In vivo and in vitro studies on the appearance of LHRH neurons in the hypothalamus of perinatal rats. Ontogenetic development of LHRH-containing neurons was studied by fluorescence and enzyme immunohistochemistry in rats. In in vitro studies, the tissues of the septal-chiasmatic and mediobasal hypothalamic areas of fetal rats on day 16.5 or 18.5 of gestation were trypsinized separately for dissociation of the neural cells, and cultured for several days. Immunopositive reaction against LHRH was first detected in nerve cells derived from both areas of the hypothalamus of the fetuses on days 16.5 and 18.5 of gestation, after 8 and 6 days culture, respectively. The cells were small, and seemed to be bipolar in morphology indicating an axon and arborized dendrites. Immunopositive material occurred in the cell soma as well as in the cellular processes. In in vivo studies, immunopositive material, possibly deposited in nerve fibers, appeared first in OVLT and simultaneously in the external layer of the median eminence of fetuses on day 20.5 of gestation. The immunoreactive fibers increased in number of both parts with development, especially after birth in the median eminence. No immunopositive material was detected within any neural cell bodies nor in the cytoplasm of any ependymal cells.


Identification by immunofluorescence of prolactin- and somatotropin-producing cells in the pituitary gland of the Mexican axolotl (Ambystoma mexicanum, Shaw). The indirect immunofluorescence procedure was used to identify prolactin (LTH)-and somatotropin (STH)-producing cells in the pituitary of the Mexican axolotl. Histological staining techniques were employed to corroborate immunocytological results. The LTH cells are large, orange-staining cells (acidophils 1) distributed in the posterior two-thirds of the pars distalis. The STH cells are small, erythrosinophilic elements (acidophils 2) principally concentrated in the dorsal part of the pars distalis.


The functional and structural border between the CSF- and blood-milieu in the circumventricular organs (organum vasculosum laminae terminalis, subfornical organ, area postrema) of the rat. The present study continues a previous investigation on the median eminence (EM) (Krisch et al., 1978). In rats with high levels of neurohormones (LHRH, vasopressin) a limited immunohistochemical labeling of perivascular tanycyte processes can be observed surrounding capillaries in the marginal region of the organum vasculosum laminae terminalis (OVLT) and in the inner part of the subfornical organ (SFO). This labeling extends from the perivascular space a short distance along the tanycyte processes. By conventional electron microscopy and by freeze-etching, tight junctions are demonstrated at a distance from the capillary lumen which corresponds to the borderline of the immunohistochemical labeling of perivascular tanycyte processes in light microscopic preparations. The tight junctions are arranged in several parallel and helical rows and correspond to those found in the median eminence. Consequently, the immunohistochemical labeling the OVLT and in the SFO marks the intercellular cleft. In the circumventricular organs the immunostaining labels the extension of the perivascular space characterized by the hemal milieu. The perivascular space is separated off by tight junctions from the CSF-milieu of the adjacent neuropil. Furthermore, the present study demonstrates tight junctions in the marginal region of the area postrema (AP) between the perivascular processes of the tanycytes.


Ultrastructure of rat pinealocytes in vitro: influence of gonadotropic hormones and LH-RH. The influence of gonadotropic hormones on the ultrastructure of rat pinealocytesin short-term organ culture was studied. Human chorionic gonadotropin (HCG), as well as pregnant mare serum gonadotropin (PMSG), caused a marked activation of pinealocytes. An hypothesis is discussed implying the presence of a feedback mechanism between the pineal organ and the hypothalamo-hypophysial system.


Structural analysis and in vitro processing to p5 rRNA of a 9S RNA molecule isolated from an rne mutant of E. coli. A temperature-sensitive mutant of E. coli, rne (RNAase E-), accumulates a number of relatively small RNA molecules at the nonpermissive temperature. One of these molecules, 9S, contains 5S rRNA sequences. The 9S RNA can be processed in vitro to 5S (p5) and a 4S-sized molecule. The 9S and the p5 and 4S RNAs processed from it were all characterized by RNAase T1 fingerprinting and pancreatic RNAase redigestion analysis. Unique oligonucleotides found in the p5 and the 4S RNA are present as unique oligonucleotides in the 9S RNA, suggesting that each 9S molecule contains one 5S and one 4S RNA moiety. Since the 9S RNA contains about 420 nucleotides, and intervening sequences are not known to exist in E. coli, the 5S an 4S RNAs cannot be more than 200 nucleotides apart in the genome. E. coli contains at least three different sequence variants of 5S rRNA, all of which can be identified in the 9S transcript, indicating that 9S RNA is transcribed from most, if not all, of the active rRNA genes, and that RNAase E processes transcripts derived from all these rRNA genes. No modified bases were found in the 4S RNA, nor was its sequence compatible with the known sequences of tRNAs found near the 5S rRNA. We therefore conclude that the 9S RNA doses not contain tRNA, and that the tRNA molecules located near the 5S rRNA are distal to it.


Relationships between fibronectin (LETS protein) and actin. Double label immunofluorescence was used to study the distribution of fibronectin (LETS protein), actin and intermediate filaments in cultured cells. No relationship was observed between fibronectin and intermediated filaments, but fibronectin and actin showed coincident staining in a large proportion of cells during spreading or when fully spread. The distributions of actin and fibronectin staining during the course of cell spreading progressed through a series of patterns. Certain actin patterns correlated with certain fibronectin patterns. When fibrillar patterns developed, there was correspondence between the two fibrillar arrays in 80--100% of the cells. These results suggest a transmembrane relationship between microfilament bundles and fibronectin. We propose that fibronectin may participate in the formation of attachment plaques and discuss the interrelationship between plaques, microfilament bundles and fibronectin in cell-substratum and cell-cell contacts.


Nonsense and insertion mutants in the relA gene of E. coli: cloning relA. We have made use of lysogens of a specialized transducing bacteriophage, lambdapyrG+ relA+, to select nonsense (relAnon) and insertion (relAins) mutations in the relA gene. Three independent relAnon mutants were isolated on the phage. In all three, the relaxed phenotype was suppressed by supD, supE, supF or sup6. Three independent relAins mutants were isolated, all containing an insertion element (probably IS2) in an apparently identical location in the relA gene. Polyacrylamide gel electrophoretic analysis of peptides synthesized by the phages in ultraviolet lightkilled host cells revealed that no stringent factor was coded for by either the relAins or relAnon phages (the latter in a sup+ cell); stringent factor was detected when the relAnon phages were used in a similar experiment with supD or supE host cells. The relAnon and relAins mutations could be crossed in haploid form in the E. coli chromosome. These recombinants grew with a normal doubling time, had a ppGpp pool which was between 70 and 100% compared with the classical relA strain, and underwent a normal carbon source shift-down. A restriction endonuclease map of the pyrG relA region of the specialized transducing phage is presented in which the position of the insertion element (recognized by a novel Hind III-cut site) defines the position of the relA gene. This position was verified by an analysis of the structure of five plasmids formed by cloning portions of the region in the pBR322 cloning vehicle. Our results indicate that the relA gene is not an essential cellular function, that there might be a second mechanism for the synthesis of basal level ppGpp in the cell and that the sole function of the relA gene is apparently the high level ppGpp synthesis triggered in response to deacylated tRNA.


The transposon Tn9 generates a 9 bp repeated sequence during integration. We performed a genetic and sequencing analysis of insertions of the transposon Tn9 into the lac operon of E. coli. Genetic mapping of 70 insertions into lacl and Z shows that starting from the same point on the chromosome, Tn9 goes to at least 50 different points in these two genes. Although there are preferred regions for insertion, these consist of multiple integration points within a small area, as demonstrated by pairwise crosses and restriction mapping. Sequence analysis of three Tn9 insertions reveals that Tn9 integration is associated with a direct repeat of 9 base pairs (bp) of host sequence. We show that these extra 9 nucleotide pairs are generated upon insertion and not brought in with the element.


Sensory adaptation mutants of E. coli.

Sensory adaptation mutants of E. coli. The ability of E. coli to adapt to constant levels of attractant and repellent chemicals was studied by examining the patterns of flagellar movement in cells subjected to abrupt concentration changes. Wild-type bacteria exhibited transient responses to such stimuli, in support of previous findings. Nonchemotactic mutants of the cheX class responded to both attractants and repellents, but were unable to terminate these behavioral changes as long as the stimulating chemical was present. The sensory adaptation defect of cheX strains may be due to an inability to methylate several cytoplasmic membrane proteins that initiate changes in flagellar movement in response to chemoreceptor signals. Based on these results, possible mechanisms of stimulus transduction and sensory adaptation during chemotaxis are discussed.


Failure of sensory adaptation in bacterial mutants that are defective in a protein methylation reaction. Chemotactic bacteria, such as E. coli, detect changes in the chemical composition of the environment. Addition of an attractant or repellent leads to an immediate response, characterized by a change in the swimming behavior of the cells--a process known as sensory excitation. However, the response gradually disappears with time, despite the continued presence of the chemical--a process known as sensory adaptation. We report here the behavior of a class of nonchemotactic mutants (cheX) that can carry out sensory excitation but are defective in the process of sensory adaptation. These mutants are also defective in the ability to carry out a protein methylation reaction which has previously been implicated in the adaptation process (Goy, Springer and Adler, 1977). The results presented here establish a firm relationship between the methylation reaction and sensory adaptation.


[Therapeutic problems in neurological bladder in the child (author's transl)]. On the basis of a retrospective review of 116 cases of neurological bladder in children, consisting essentially of patients with spina bifida and meningomyelocoele, the authors consider their present attitude to this problem. They take a simple neurological classification as a basis, and the various forms of treatment (cutaneous ureteric transplantation, physiotherapy, divulsion, sphincterotomy, anti-reflux plastic procedures, treatment of incontinence) are reviewed in terms of method, indications and results.


Practical use of swan-type double-bending gastrofiberscope. After devising remote-controlled angle to gastrofiberscope in 1964, the authors invented a swan-type double-bending gastrofiberscope in 1967. The double-bending gastrofiberscope does not require any change in a patient's position, for the motion of the tip is sufficient to observe every part of the stomach on supine position. In order to relax the patient both physically and mentally, relaxation of the patient is obtained by taking off this excessive attention on the endoscopic examination. Clear visual field can be obtained at the moment when the fiberscope passes the cardia, by stopping the scope just above the cardia and inflating sufficient air, and then, inserting the scope with the lens directed to the anterior wall of the greater curvature. Several marks are put on the body of the scope, as a functional scale, which makes its handling easier.


Longitudinal serological study of malaria in infants in the West African savanna. Comparisons in infants exposed to, or protected from, transmission from birth. Two infant populations, the one exposed to intense malaria transmission and the other protected, were followed and compared by six serological tests. The IgG and IgM levels increased with age and were systematically, though only slightly, lower in the protected children. The results of three Plasmodium falciparum tests (precipitin, indirect fluorescent antibody (IFA) and indirect haemagglutination and one P. malariae test (IFA) were high at birth and decreased rapidly afterwards in both populations. In the unprotected population, this decrease was followed by an increase, closely associated with the parasitological findings, while in the protected population the decrease continued to very low levels.


Further epidemiological evaluation of a malaria model. The malaria model previously fitted to 1 year of baseline data from the Garki District in the Sudan savanna of northern Nigeria was tested against data collected in the same area over a period of 3 years, including 1(1/2) years during which the insides of houses in certain villages were sprayed with propoxur. It was also tested against data collected in Kisumu, Kenya, also over a period of 3 years, including 20 months during which the insides of houses in part of the area were sprayed with fenitrothion. The test consisted in using the vectorial capacity, calculated from the entomological observations made in the above places and periods, as input in the Garki model while keeping the other parameters as fitted to the Garki baseline data, and in comparing the prevalence of Plasmodium falciparum parasitaemia as estimated by the model to that actually observed. There was relatively good agreement and the model is considered epidemiologically satisfactory and fit for use in planning malaria control operations.


Changing patterns in the humoral immune response to malaria before, during, and after the application of control measures: a longitudinal study in the West African savanna. A longitudinal seroimmunological investigation of malaria was performed as part of the WHO research project conducted in the northern part of Nigeria from 1970 to 1975. The project included a preintervention phase, an intervention phase with application of malaria control measures (spraying of residual insecticide and mass drug administration), and a postintervention phase. Serological observations were made on the total population of eight villages consisting of approximately 3000 persons. Six immunological parameters were studied, namely, the serum levels of IgG and IgM, the number of bands of precipitation for Plasmodium falciparum in the double diffusion (Ouchterlony) test, the titres of antibodies for P. falciparum and P. malariae in the indirect fluorescent antibody (IFA) test, and the titres of agglutinating antibodies for P. falciparum by the indirect (passive) haemagglutination (IHA) test. The serological results were used to evaluate the impact on the humoral immune response of different levels of parasitaemia resulting, in the unprotected population, from natural factors such as seasons and ageing and in the protected population, from human intervention through the application of control measures and their interruption. The linkage by computer processing of the longitudinal data allowed analysis of the relationship between the results of a serological test in the same person at different surveys, and analysis of correlation between serological results and the concurrent parasitological findings. The correlation between parasitaemia and the results of the different serological tests at the same survey in the same person were also examined and analysed in terms of sensitivity and specificity of the tests.


A controlled field trial of plain and aluminium hydroxide-adsorbed cholera vaccines in Surabaya, Indonesia, during 1973--75. A controlled field trial comparing the effectiveness of a plain cholera vaccine with that of a vaccine adsorbed to aluminium hydroxide was carried out in a cholera-endemic area of Indonesia during 1973-75. Tetanus toxoid adsorbed to aluminium phosphate was used as the control. In vaccinees aged 1-4 years, the adsorbed cholera vaccine provided about 88% protection for 6 months following vaccination and still provided about 50% protection between 11 and 14 months after vaccination. In the same age group, the plain vaccine provided only 53% protection during the first 6 months and no appreciable protection beyond that period. In those aged 5 years and over, both vaccines provided 50-60% protection throughout the period of observation (14 months). Neither vaccine caused any serious side effects.


Detection of anti-HBs antibody by means of an indirect micro-ELISA method. The Hepanostika micro-ELISA System was used, with some modifications, for the indirect detection of anti-HBs antibodies. The proposed method is rapid, easy to perform, reproducible, and specific and approaches radioimmunoassay in sensitivity (92.6% agreement). An advantage over radioimmunoassay is the possibility of revealing antibodies in sera containing immune complexes, but this will require further investigation.


Effects of verapamil on myocardial performance in coronary disease. Verapamil, a calcium antagonist, has been used extensively for treatment of cardiac arrhythmias. Concern persists, however, that it may seriously depress myocardial function in cardiac patients. To investigate this possibility, 20 patients with coronary artery disease (CAD) but no heart failure were given intravenous verapamil (0.1 mg/kg bolus, followed by 0.005 mg/kg/min infusion), and studied hemodynamically and angiographically. Verapamil markedly lowered mean aortic pressure (94 +/- 17 to 82 +/- 13 mm Hg, p less than 0.0005) and systemic vascular resistance (1413 +/- 429 to 1069 +/- 235 dyn-sec-cm5, p less than 0.0005). Simultaneously, all indices of left ventricular (LV) performance greatly improved: cardiac index rose from 2.8 +/- 0.6 to 3.1 +/- 0.7 1/min/m2 (p less than 0.0005), mean velocity of circumferential fiber shortening increased from 0.85 +/- 0.39 to 0.97 +/- 0.46 circ/sec (p less than 0.01), and ejection fraction improved from 55 +/- 16 to 61 +/- 18% (p less than 0.01). No significant changes were noted in the heart rate before and after verapamil administration, and verapamil did not worsen the extent of LV asynergy in the majority of patients. In patients with CAD, the intrinsic negative inotropic effect of verapamil is of negligible importance because its potent vasodilatory properties more than compensate for any intrinsic decrease in LV contractility, and thereby improve the overall cardiac function.


Effect of prazosin vs placebo on chronic left ventricular heart failure. The effect of the vasodilator prazosin vs placebo on exercise duration until marked dyspnea, and on left ventricular function measured by echocardiography, was evaluated in a double-blind, randomized study in 24 patients with chronic left ventricular failure despite digitalis and diuretic therapy. Compared with the double-blind placebo, prazosin reduced resting systolic and diastolic blood pressure and systolic blood pressure times heart rate, improved clinical symptoms, decreased cardiothoracic ratio measured by chest roentgenography, decreased left ventricular and left atrial dimensions, improved ejection fraction and Vcf measured by echocardiography, and improved treadmill exercise duration. All 12 patients taking prazosin had greater than or equal to 20% improved treadmill exercise duration; none of 12 receiving placebo improved. In six of 12 patients taking prazosin, roentgenographic evidence of pulmonary venous congestion disappeared compared with none of the patients on placebo. These data suggest that prazosin may be effective in treating chronic left ventricular failure.


Assessment of the EMIT-(TM) technique as a screening test for opiates and methadone for a methadone maintenance clinic and its calibration by Bayesian statistics. 1. The performance of the semi-quantitative enzyme multiplied immunoassay technique (EMIT, Syva Corp.) for opiates and methadone has been examined as a screening procedure of urine samples from a methadone maintenance programme. The predictive value model that is based upon Bayesian statistics was used to determine screening levels for the EMIT assays. 2. With a predictive value of a negative result of 100%, the EMIT opiate assay can be used to show the absence of the indicated drugs, while positive results can be confirmed by a non-immunological technique. A screening level of 1.0 micrograms/ml for the opiate assay, conforms to this model. 3. The EMIT methadone assay was shown to have a predictive value of a negative result of 28% with respect to thin-layer chromatographic (TLC) results. This discrepancy between EMIT and TLC can not be explained by sensitivity alone. The manufacturer's recommended 0.5 micrograms/ml cutoff has been used therefore for the methadone assay.


Enzyme immunoassay of carbamazepine with a centrifugal analyzer. We describe a rapid enzyme immunoassay for carbamazepine with a centrifugal analyzer (Rotochem IIA-36). Reagent costs are reduced fourfold while good precision and sensitivity are maintained. Sample volume is 10 microliter, and as many as 28 patients' sera can be measured during an assay time of 225 s. Assay temperature is 30 degrees C, the wavelength 340 nn. Linearity is excellent for a carbamazepine concentration range of 1 to 12 mg/L; analytical recovery is quantitative. Results correlate well with those by liquid- and gas-liquid chromatography. Absorbance rates for each carbamazepine concentration are acquired by a multi-point kinetic rate program and a computer program provides a logit-log transformation of absorbance rate vs. concentration data for final calculations in the assay. Hemoglobin interference precludes analysis of severely hemolyzed specimens.


An "antibody electrode," preliminary report on a new approach in enzyme immunoassay. We report preliminary experiments on a new, sensitive, and reliable procedure for enzyme immunoassay of various antigens in biological fluids. The method, developed from the biological model "hepatitis B surface-antigen/antibodies," is less time consuming than most immunochemical techniques and eliminates many inconveniences arising from use of isotopes. We use a solid-phase "sandwich" procedure, the antibodies being immobilized on gelatin membranes, and determine antigen concentration with the help of an iodide-sensitive electrode modified by fixing the active membrane onto the crystal sensor. We have established the analytical criteria of the method and compared it with the solid-phase radioimmunoassay for the surface antigen in dilution series. One-tenth microgram of antigen per liter can be reproducibly detected with our method. Use of antibody electrode should easily be extended to assay of other antigens and haptens that usually are determined by radioimmunoassay.


Transverse maxillary growth in combined cleft lip and palate. A longitudinal roentgencephalometric study by the implant method. Transverse maxillary growth was studied radiographically by the metallic implant method in 5 Danish boys with combined cleft lip and palate. The study covered the age range of five to 20 years with individual observation periods of 9 to 12 years. Lip closure and palate closure had been performed at two months and 24 months of age respectively by conventional methods. Orthodontic treatment was carried out in all cases. The maxillary width was determined from posteroanterior cephalometric radiographs as the distance between metallic implants inserted at the lower aspect of the maxillary-zygomatic process on both sides. Other roentgencephalometric measurements were obtained for further estimate of the facial growth in width. Distance curves and velocity curves for these measurements and for body weight were constructed and compared. The growth curves for maxillary width differed definitely from normal and indicated that the spontaneous transverse palatal growth was inhibited in these patients.


The urinary excretion of orotic acid and orotidine, measured by an isotope dilution assay. Unknown concentrations of orotic acid can be measured by competition with a known amount of [carboxyl-14C]orotic acid for reaction with a limiting amount of phosphoribosylpyrophosphate in the presence of orotate phosphoribosyltransferase and orotidine monophosphate decarboxylase. The dilution of the specific radioactivity in the product 14CO2 is a sensitive and accurate measure of the amount of orotic acid present in the sample. Orotidine can also be determined after hydrolytic cleavage to orotic acid. The method was used to measure orotic acid and orotidine in urine samples from newborns, healthy controls and patients with gout or deficiency of hypoxanthine-guanine phosphoribosyltransferase receiving allopurinol. Urinary excretion of orotic acid and orotidine in newborns was similar whether the infants were breast-fed or received milk powder. The excretion of orotidine was increased in all patients receiving allopurinol. After allopurinol administration orotic acid excretion was increased in gouty patients but close to normal values in patients with deficiency of hypoxanthine-guanine phosphoribosyltransferase. The results are discussed in relation to the mechanism by which allopurinol inhibits pyrimidine metabolism.


The significance of pure diffuse mesangial proliferation in idiopathic nephrotic syndrome. The prognostic significance of the finding of diffuse mesangial proliferation (DMP) in patients presenting with idiopathic nephrotic syndrome (INS) has not been well established. The clinical course, therapeutic response and final outcome of 38 patients in whom renal biopsy showed DMP are reported. They have been subdivided into 2 groups according to the absence (18 patients: group I) or presence (20 patients: group II) of superimposed lesions of focal and segmental sclerosis and/or hyalinosis (FSS/H). Clinical presentation was similar in both groups although patients in group I were less severely affected. Non of the patients of group II responded to corticosteroids, whereas in group I 2/16 responded and 2 infants remitted without treatment. At the latest assessment, 5/18 patients in group I and 10/20 in group II had progressed to terminal renal failure or had impaired renal function. Five patients in group I and 3 in group II were in clinical remission. Eight of 11 repeat biopsies performed in patients of group I showed the development of FSS/H. Thus patients with DMP seem prone to develop lesions of FSS/H. Their course if often worse than that of minimal change with FSS/H since 7 of the 10 patients who developed renal failure did so within 3 years of onset. The finding of DMP in a patient with idiopathic nephrotic syndrome is usually but not invariably an ominous feature.


Renal histology and immunopathology in distal renal tubular acidosis. Renal biospy studies are reported from 10 patients with distal renal tubular acidosis (DRTA). On the biopsies from 6 patients who had associated immunological abnormalities immunofluorescent studies for immunoglobulins, complement, and fibrin were performed. Interstitial cellular infiltration and fibrosis were common findings in patients with and without immunological abnormalities, and were usually associated with nephrocalcinosis and/or recurrent urinary infection. No immune deposits were demonstrated in association with the renal tubules. This study shows that DRTA in immunologically abnormal patients is not caused by tubular deposition of antibody or immune complexes. The possibility of cell mediated immune damage is discussed.


Bone mineral content in renal transplant patients. Bone mineral content (BMC) in the forearm was measured in 158 renal transplant patients to elucidate the relationship between bone mass and the development of bone lesions following renal transplantation. In renal transplant patients with aseptic necrosis of bone and with spontaneous fractures the BMC was significantly reduced compared with age and sex matched renal transplant control patients. Other factors connected with low BMC were age, sex, prolonged dialysis prior to transplantation, post-transplant hypercalcemia and elevated serum alkaline phosphatase at the time of the study. No relation between BMC and duration of immunosuppressive treatment was demonstrated.


Interstitial immunofluorescence in nephritis of Sjögren's syndrome. A patient with Sjögren's syndrome (SS) developed mild renal insufficiency without significant proteinuria; the diagnosis of interstitial nephritis secondary to SS was made on the basis of history, serological data and renal biopsy. The renal biopsy revealed immunofluorescence of the interstital infiltrates and the importance of this finding in the differential diagnosis of interstitial nephritis is considered. In addition, further studies to detect anti-renal antibodies and circulating immune complexes are reported and the possible interpretation of the results is discussed in regard to pathogenesis.


The long-term prognosis of patients with focal segmental glomerulosclerosis. Forty consecutive patients whose biopsies showed focal and segmental sclerosis were studied for 6 to 16 years to establish the long-term prognosis of this group of patients. By the end of a decade 21 were dead, on regular dialysis or transplanted; only one death was unrelated to renal failure. A further 8 patients had a GFR of less than 60 ml/min/1.73 m2. Only 11 patients had normal renal function, and of these eleven only four had no urinary abnormality. Actuarially calculated survival was 75% at 5 years, 50% at 10 years, and 38% at 15 years. There was no difference between the 28 adults and the 12 children in terms of evolution. Patients with a nephrotic syndrome at presentation had a poorer prognosis than those never nephrotic. Twenty nephrotic patients were treated with prednisone, and 14 of these with cyclophosphamide in addition. One patient responded with loss of proteinuria within two months to both drugs, and another lost proteinuria when treated with cyclophosphamide. Thirteen patients received allografted kidneys; a nephrotic syndrome recurred in three patients, with histological evidence of recurrent disease in two.


Allograft renal-artery stenosis: increased peripheral plasma renin activity as an early indicator of uncontrollable hypertension. We measured indices of the renin-aldosterone system and body-fluid spaces in 11 adolescents who had received a renal transplant after removal of their own diseased kidneys. None had hypervolemia but 6 had hypertension. Renal angiography revealed greater than 50% luminal occlusion by allograft renal-artery stenosis (RAS) in only the 3 patients who had severe hypertension refractory to conventional medical therapy. Excessive peripheral plasma renin activity (PRA) distinguished these patients from those who had less severe stenosis or normal angiogram, and diuretic stimulation heightened the PRA differences. We conclude that significant allograft RAS does not necessarily act like a typical single-kidney Goldblatt model until after volume depletion. Our findings indicate that peripheral PRA values can be used to assess the degree of graft ischemia clinically. This permits early identification of patients who have severe RAS that probably will be difficult to control medically, and, therefore, should be followed closely with a view of reconstructive vascular surgery before further deterioration of renal function.


The long-term effects of spinal fusion on the sacroiliac joints and ilium. Ninety-six patients who had lumbar disk excision and primary posterior fusion were studied 10 or more years after their operations. Thirty-seven per cent complained of persistent graft donor site pain. A comparison of fused patients with and without donor site pain and 36 patients who underwent simple disk excision, revealed no differences in the flexion-extension mobility of the sacroiliac joints, or degenerative changes in the sacroiliac joints. Ectopic bone formation at graft donor sites, and cluneal nerve neuromata did not influence the result. Patients with graft donor site pain had significantly greater complaints of persistent low back pain, postoperative leg pain, and lost more time from work. If the graft was taken from the same side as that of preoperative leg pain, persistent complaints were more common. It is concluded that the sacroiliac joints are not adversely affected by lumbar spine fusion, and that persistent donor site pain is more likely part of a total pain complex referred from the lumbar spine. The sacroiliac joints appear to be relatively noncontributory to problems following lumbar disk surgery.