Mechanism of ethanol-induced changes in lipid composition of Escherichia coli: inhibition of saturated fatty acid synthesis in vivo. The in vivo effects of ethanol on lipid synthesis in Escherichia coli have been examined. Under conditions which uncoupled fatty acid synthesis from phospholipid synthesis, ethanol decreased the amount of saturated fatty acids synthesized but had little effect on the selectivity of their incorporation into phospholipids. In the absence of fatty acid degradation and unsaturated fatty acid synthesis, E. coli was still able to adapt its membrane lipids to ethanol, while the inhibition of total fatty acid synthesis eliminated this response. During growth in the presence of ethanol, strain K1060 (an unsaturated fatty acid auxotroph) incorporated an increased amount of exogenous heptadecanoic acid (17:0) to compensate for the reduction in palmitic acid (16:0) available from biosynthesis. Thus, our results indicate that the reduced levels of saturated fatty acids observed in the phospholipids of E. coli following growth in the presence of ethanol result primarily from a decrease in the amounts of saturated fatty acids available for phospholipid synthesis.


Stereochemistry of reactions catalyzed by glutamate decarboxylase. When the decarboxylation of L-glutamic acid by the glutamate decarboxylase from Escherichia coli is carried out in D2O, the product gamma-aminobutyric acid contains a single deuterium atom. The stereochemistry of this material was established by conversion to levorotatory methyl 4-phthalimido [4(-2)H] butyrate. The dextrorotatory isomer of the latter compound was synthesized from S-[2(-2)H] glycine by a series of reactions not affecting the stereochemistry at the chiral center. Thus, the decarboxylation of glutamic acid occurs with retention of configuration. Decarboxylation of L-alpha-methylglutamic acid by this enzyme produced levorotatory gamma-aminovaleric acid and thus also occurs with retention of configuration.


Shape of protein L11 from the 50S ribosomal subunit of Escherichia coli. Protein L11 from the 50S ribosomal subunit of Escherichia coli A19 was purified by a method using nondenaturing conditions. Its shape in solution was studied by hydrodynamic and low-angle x-ray scattering experiments. The results from both methods are in good agreement. In buffers similar to the ribosomal reconstitution buffer, the protein is monomeric at concentrations up to 3 mg/mL and has a molecular weight of 16 000-17 000. The protein molecule resembles a prolate ellipsoid with an axial ratio of 5-6:1 a radius of gyration of 34 A, and a maximal length of 150 A. From the low-angle x-ray diffraction data, a more refined model of the protein molecule has been constructed consisting of two ellipsoids joined by their long axes.


Spreading of biomembranes at the air/water interface. This paper presents the compression isotherms obtained by spreading membranes of intestinal brush border, human erythrocyte and Escherichia coli (cytoplasmic) at the air/water interface. Unilamellar membrane films were formed, with a good yield, at zero surface pressure, whereas multilamellar structures were formed at high surface pressure. Once formed, the films were particularly stable and could be manipulated without any detectable loss. With doubly-labelled E. coli cytoplasmic membrane, we could show that phospholipids and proteins spread, with the same yield, as a single unit. Moreover, we studied the influence of hydrolytic enzymes, chemical agents and cations on the compression isotherm of biomembranes. The resultant changes in architecture of membrane films can provide a very simple method of studying the influence of membrane packing on catalytic activity and protein conformation of membrane-bound proteins.


Synthesis of alpha3 phage DNA in replication mutants of Escherichia coli. Host functions for DNA replication of bacteriophage alpha3, a representative of group A microvirid phages, were studied using dna and rep mutants of Escherichia coli. In dna+ cells, conversion of phage alpha3 single-stranded DNA (SS) into the double-stranded replicative form (RF) was insensitive to 30--150 microgram/ml of chloramphenicol, 200 microgram/ml of rifampicin, 50 microgram/ml of nalidixic acid, or 200 microgram/ml of novobiocin. At 43 degrees C, synthesis of the parental RF was inhibited in dnaG and dnaZ mutants, but not in dnaE and rep strains. Replication of phage alpha3 progeny RF was prevented by 50 microgram/ml of mitomycin C (in hcr+ bacteria), 50 microgram/ml of nalidixic acid or 200 microgram/ml of novoviocin, but neither by 30 microgram/ml of chloramphenicol nor by 200 microgram/ml of rifampicin. Besides dnaG and dnaZ gene products, dnaE and rep functions were essential for progeny RF synthesis. Host factor dependence of alpha3 was relatively simple and, in contrast with phages phiX174 and G4, alpha3 did not require dnaB and dnaC(D) activities.


The efficiency of methionine incorporation from isoaccepting species of tRNAMet into rabbit globin in an homologous reticulocyte lysate system. Rabbit globin alpha and beta chains were labeled with [3H]leucine, and with [35S] -methionine from reticulocyte tRNAMet isoacceptors using a rabbit reticulocyte cell-free synthesis system. [35S]Methionine from the three tRNAMet species isolated by RPC-5 chromatography was incorporated into internal positions of both alpha and beta globin. The initiator tRNA, tRNAIMet, exhibited very low efficiency for incorporating methionine internally, while tRNAIIMet was four times more efficient than tRNAIIIMet. Amino acid analysis of the tryptic peptides of the labeled globins revealed that all three isoacceptors incorporated methionine into the normal methionine peptides. Similar studies with Escherichia coli [35S]Met-tRNAfMet showed a 3-fold increase over the reticulocyte initiator tRNA in its capacity to incorporate methionine into the internal positions of rabbit globin.


On the accessibility and selection of the initiator site of mRNA in protein synthesis. The specificity of the cell-free system of Escherichia coli for mRNA was examined, and the "accessibility" of some natural and synthetic RNAs to the ribosomes was determined by measurement of AcPhe-tRNA and fMet-tRNA binding, AcPhe-puromycin and fMet-puromycin formation, and polypeptide synthesis. The E. coli system effectively initiates the translation of various synthetic RNAs with AcPhe-tRNA or fMet-tRNA under conditions optimal for the translation of viral RNA. Poly(A,G,U) is accessible to the ribosomes according to all of the above criteria. Poly(A,C,G,U), 23 S rRNA, R17 RNA, and MS2 RNA, on the other hand, show limited accessibility when tested for initiator tRNA binding, or for AcPhe-puromycin and fMet-puromycin formation. MS2 and R17 RNA, but not poly(A,C,G,U) and 23 S rRNA, show accessibility when measured by polypeptide synthesis. The results suggest that, except at initiator sites of natural mRNA, an RNA containing about equal amounts of all four bases is inaccessible to E. coli ribosomes for polypeptide synthesis. Rate constants obtained for fMet-tRNA binding with MS2 RNA, poly(A,G,U), and poly(C,G,U) indicate that the ribosomes do not have any special affinity for the viral RNA. Thus, the selection of the initiator site in protein synthesis may be critically determined more by the accessibility of the initiator codon than by ribosomal recognition of the site.


Covalent cross-linking of ribosomal RNA and proteins by methylene blue-sensitized photooxidation. Ribosomal proteins are covalently cross-linked to ribosomal RNA by irradiation with visible light in the presence of methylene blue and O2. Proteins S3, S4, S5 and S7 from the 30 S subunit of Escherichia coli ribosomes and L2 and L3 from the 50 S subunit are among the cross-linked proteins. S3 and S5 had not previously been identified as RNA-binding proteins.


Purification of RNAase II by preparative polyacrylamide gel electrophoresis. Purification of RNAase II to electrophoretic homogeneity is described. The exonuclease is activated by K+ and Mg2+ and hydrolyses poly(A) to 5'-AMP, exclusively as described by Nossal and Singer (1968, J. Biol. Chem. 243, 913--922). To separate RNAase II from ribosomes, DEAE-cellulose chromatography was used. Two additional chromatographic steps give a preparation that yields 10 bands after analytical polyacrylamide gel electrophoresis. Preparative polyacrylamide gel electrophoresis resulted in a final preparation which on analytical polyacrylamide gels gives a single band. A molecular weight of 76 000 +/- 4000 was obtained from Sephadex G-200 chromatography, with three bands from sodium dodecyl sulfate (SDS) denaturation and SDS gel electrophoresis. The subunits have a molecular weight of 40 000 +/- 2000, 33 000 +/- 2000, and 26 000 +/- 1000. The enzyme thus appears to consist of three dissimilar subunits.


Factors influencing the observed half-lives of specific synthetic capacities in Saccharomyces cerevisiae. We have identified a variety of factors affecting the stability of allophanate hydrolase-specific and gross cellular protein synthetic capacities. These synthetic capacities have been extrapolated by many laboratories to represent functional messenger RNAs. Synthetic capacity turnover rates that we measured were greater in diploid organisms than in haploid strains and were proportional to the temperature of the culture medium. The stability of allophanate hydrolase-specific synthetic capacity was not influenced by alterations in the nitrogen source provided in the culture medium, but was increased up to 15-fold by the total inhibition of protein synthesis. Cultures in which protein synthesis was inhibited as little as 20% exhibited hydrolase-specific synthetic capacities more than 2-fold greater than those observed in the absence of inhibition.


Bacteriophage phiX174 growth in an Escherichia coli dnaIts mutant, KS810. A bacteriophage phiX174-sensitive Escherichia coli dnaIts mutant, KS810, was constructed and growth of phiX174 in the cells was investigated. phiX174 and phiX174am3trD could grow normally at 43 degrees C as well as 27 degrees C, therefore we conclude that the growth of bacteriophage phiX174 is not dependent upon the host dnaI gene product.


On the mechanism of action of Escherichia coli tryptophan synthase. Steady-state investigations. Tryptophan synthase from Escherichia coli (L-serine hydro-lyase (adding indole), EC 4.2.1.20) synthesizes L-trypotophan from indoleglycerol phosphate and L-serine, releasing glyceraldehyde 3-phosphate, or from indole and L-serine. The latter reaction (B reaction), catalyzed either by the beta2 species or by the (alpha2 beta2) complex, has been studied by steady-state methods. A sequential mechanism is indicated. Inhibition experiments with the substrate analogue benzimidazole were carried out in order to distinguish between random and ordered mechanisms. The results are compatible with a random sequential mechanism. The dissociation constants of the enzyme-substrate complexes are evaluated. When catalyzed by the tetrameric complex (alpha2 beta2) the B reaction is inhibited by higher concentrations of the substrate indole. This inhibition does not follow the usual substrate inhibition pattern. The question whether the binding of indole to the alpha-subunit exerts an inhibitory effect on the beta2 species, possibly by reversing the activation by the alpha subunit of the beta2 species, is discussed.


Intermolecular interaction studies on native and enzyme-treated acid-soluble collagen. The aggregative properties of acid-soluble collagen, native or enzyme treated, have been studied by electric birefringence and low shear rate viscosity. A unique type of aggregate has been found, about 700 nm long for native collagen and 530 nm for pepsin treated, regardless of the acetic acid concentration in the range 1--100 mM. The number of aggregates increases with collagen concentrations, as could be expected for electrostatic interactions. On the contrary, pepsin-extracted cartilage collagen forms aggregates of covalent nature, the number of which is independent of concentration. Viscosity measurements show two different interaction mechanisms: a short distance one which can be identified with the electric birefringence-detected aggregation, and a long distance superstructure which disappears when salt is added to the solution.


Rotational diffusion of Escherichia coli ribosomes. I. - Free 70 S, 50 S and 30 S particles. The rotational brownian diffusion of Escherichia coli ribosomes has been studied by following the transient dichroism generated by optical excitation of a covalent probe into its triplet state. The induced absorption anisotropy decays exponentially with characteristic correlation times: 2.5 microseconds, 1.6 microseconds and 1.1 microseconds for the 70S ribosome and the 50S and 30S subparticles respectively. The corresponding Stokes radii are in the same order, 133 A, 115 A and 103 A. The hydrodynamic properties are discussed in terms of an ellipsoidal shape of the ribosome particles.


Rotational diffusion of Escherichia coli ribosomes. II. - Ribosome attachment to polyurydilic acid. Ribosome attachment to poly(U) has been studied by following the rotational diffusion of polyribosomes in solution. On the average, 13-17 and 50 nucleotides are found to be associated with 30S and 70S ribosome respectively. For an equal length of poly(U), the number of particles in a 30S polysome is four times that in a 70S polysome. The results are consistent with a structure of the polysome in which individual ribosomes are in close contact.


[Biosynthesis of thiazole from thiamine in Escherichia coli]. The incorporation into the thiazole moiety of thiamine of several labeled compounds has been studied on short time incubations of washed-cells suspensions. No incorporation of radioactivity from [G-14C] methionine was found in a mutant auxotrophic for methionine. No radioactivity was incorporated from [U-14C] aspartate or from [U-14C] serine. The incorporation of 35S from sulphate was lowered by cysteine or glutathione but was unaffected by methionine or homocystine. Although the synthesis of thiazole is dependent on methionine, neither the sulphur atom nor the carbon chain of thiazole originate from methonine in E. coli. No carbon originates from cysteine which is the likely direct donor of sulphur.


[Dependence of inhibition by levorin of amino acid incorporation into protoplasts and subcellular proteins of Candida albicans on the change of endocellular potassium concentration]. Levorin is found to decrease more efficiently potassium concentration in C. albicans protoplasts under their incubation in the presence of sodium than in the medium containing the equivalent amount of potassium. Minimal inhibitory concentration of levorin for resistant C. albicans cells incubated on potassium-depeleted medium was in 4 times lower than for cells incubated in potassium-enriched medium. The decrease of membrane permeability for 14C-amino acids and their incorporation into membrane, ribosomal and soluble proteins under the effect of levorin was more pronounced when protoplasts were cultivated in sodium-containing medium than in potassium-containing one. In both media the inhibition of 14C-amino acid incorporation by levorin into ribosomal and cytosol proteins was more efficient than into membrane proteins, but these differences were less pronounced in case of potassium-containing medium.


[Determination of leucine-binding protein content of Escherichia coli cells]. A radioimmunochemical method of determination of leucine-binding protein and a method, based on the selective absorption of the protein and its complex with leucine on DEAE-cellulose, has been developed. The protein content in the E. coli cells at different stage of growth has been determined by the radioimmunochemical and equilibrium dialysis methods. It was shown that the protein content in the cells is practically independent of the growth-phase.


[Translation of poly U by ribosomes from rel+ and rel- E. coli strains]. A translation of polyU in a cell-free system from CP78 (relA+) and CP79 (relA-) E. coli strains is investigated. The strains studied no not differ in misreading at Mg2+ concentration of 20 mM and concentrations of 16 mM (optimal for phenylalanine incorporation) and 18-22 mM (optimal for leucine incorporation) respectively. It is found that leucine incorporation increases similarly in both strains under conditions simulating an amino acid deficience (in phenylalanine-free incubation medium): the ratio leucine(-phenylalanine)/leucine (+phenylalanine) is 3.5-4 at a concentration of enzymatic fraction protein being 15-30 mkg per example, and it is 2 st a concentration of enzymatic fraction of 60-180 mkg of protein. It is suggested that differences in activities of a number of enzymes under amino acid starvation in Rel+ and Rel- cells are not due to differences in the precision of mRNA translation, but depend on the activity of respective genes.


Effects of diazepam on phobic avoidance behavior and phobic anxiety. The efficacy of diazepam for controlling phobic avoidance behavior and phobic anxiety and the potential usefulness of the behavioral approach measure for screening new antianxiety drugs was investigated in a double-blind study. The distance from the phobic object beyond which the subject would not approach and a rating of subjective anxiety at closest approach were made in 14 phobic patients immediately before and again 2 hr after 10-mg oral diazepam or placebo. Behavioral approach to the object was increased and subjective anxiety was decreased by diazepam as compared to placebo. The method appeared useful for early clinical screening of new drugs because it discriminated drug from placebo in a single short session using small groups and because it employed a more objective dependent measure than self-report of subjective feeling states.


A phase II clinical trial of cytembena. Clinical screening group of E.O.R.T.C. In a phase II clinical (screening) trial, Cytembena has induced 20% objective regressions in uterus cervix carcinoma. This agent did not induce any myelosuppressive toxicity.


The interpretation of the mass spectrum of an ornithine-containing lipid from Thiobacillus thiooxidans. The electron impact mass spectrum of a previously identified ornithine-containing lipid from Thiobacillus thiooxidans has been interpreted using exact mass measurements, low and high energy ionization, and defocused metastable studies. The spectrum, which did not contain a molecular ion for the intact lipid, was consistent with cyclization of the ornithine zwitterionic moiety with elimination of water to give 3[3'-(11,12-methylene-2-hydroxyoctadecanoxy)hexadecanylamine]-2-piperidone. Production of this sufficiently volatile species for mass spectral analysis was accomplished by gentle pyrolysis in the mass spectrometer source. The spectrum can be understood to arise by three primary decompositions which serve to separate the two fatty acid constituents. The remainder of the spectrum is consistent with the expected fragmentations of these constituents.


[Use of 51Cr-labelled macrophages for the objective evaluation of macrophage migration inhibition]. A possibility of using 51Cr-labeld macrophages migrating from the capillaries and lysed with sodium dodecyl sulfate for objective and quantitative assessment of the results of macrophage migration inhibition (MMI) microassay was demonstrated on the basis of correlation analysis of the experimental data. Migration area and sensitivity of 51Cr-labeled macrophages to the action of the factor of migration inhibition were found not to differ from those of unlabeled macrophages. In the assessment of the absolute macrophage migration values and its reduction under the action of the factor of migration inhibition formed in the H-2 system, and also of the specificity of this reduction there was revealed a high correlation between the migration area and the value of the migrant cell label.


Multimarker analysis of T-cell chronic lymphocytic leukemia. A 68-yr-old male with chronic lymphocytic leukemia (CLL) presented with splenomegaly and skin infiltration but no lymphadenopathy. The peripheral blood WBC cound was 300 x 10(9)/liter, with 95% small mature-appearing lymphocytes that were E-rosette positive and EAC-rosette negative. Further characterization of the patient's cells was performed using antisera with known lymphoid sub-population specificity. Anti-p23,30, which reacts with normal circulating B cells but not with T cells or thymocytes, was unreactive with the patient's cells. Anti-311, which reacts with both thymocytes and circulating T cells, was reactive with the patient's cells. Anti-Bk, which reacts only with thymocytes and not with circulating T-cells, failed to react with the patient's cells. The enzyme terminal deoxynucleotidyl transferase, present in thymocytes but absent for circulating T-cells, was also absent from the patient's lymphoid cells. Multimarker analysis therefore showed a mature T-lymphocyte phenotype on this patient's leukemia cells. Further functional analysis will probably show that such cells represent clonal expansion of a mature T-cell subpopulation, analogous to the B-cell clonality of common-variant CLL.


Pediatrics in an eighteenth century remedy book. A manuscript home remedy book of several hundred pages in German, with entries from 1631 to 1861, was found in the Washington University School of Medicine Library some years ago. Two sections on diseases of children, from the early seventeenth to the mid-eighteenth centuries, have been translated and compared with both printed and manuscript pediatric works. People cited and geographicl places mentioned are discussed. The final conclusion reached is that the text is closer in tone and philosophy to the stream of manuscripts going back to classical times than it is to the printed book of its time. An appendix lists treatment of the wet-nurse in place of the child, and treatments for debility or wasting and pain in the abdomen, and traces these through classical and later manuscript sources.


MEDLEARN: a computer-assisted instruction (CAI) program for MEDLARS. *MEDLEARN*, a second-generation computer-assisted instruction (CAI) program available (nationally) since October 1976, provides on-line training for MEDLINE, one of the National Library of Medicine's (NLM) Medical Literature Analysis and Retrieval System (MEDLARS) data base. *MEDLEARN* was developed as a joint effort between NLM and The George Washington University Medical Center. Using MEDLINE formats throughout, *MEDLEARN* combines tutorial dialogue, drill and practice, testing, and simulation. The program was designed in three tracks oriented to basic methods, advanced techniques, and new developments. Each topic is presented on two levels, permitting an alternate explanation for users encountering difficulty. *MEDLEARN*, coded in the computer language PILOT, was developed with a modular structure which promotes ease of writing and revision. A versatile control structure maximizes student control. Frequent interactions check immediate recall, general comprehension, and integration of knowledge. Two MEDLINE simulations are included, providing the student an opportunity to formulate and execute a search, have it evaluated, and then perform the search in MEDLINE. Commenting, news broadcasting, and monitoring (with permission only) capabilities are also available. Subjective field appraisals have been positive and NLM plans to expand *MEDLEARN* and produce similar programs for other data bases.


A description of the Kurzweil reading machine and a status report on its testing and dissemination. This paper presents a technical overview of the different subsystems of the Kurzweil Reading Machine--the nature of the information processed by each subsystem and the type of transformations performed. A description of how the user directs the machine is provided. The Veterans Administration's testing program, as well as those of the Bureau of Education for the Handicapped, the Rehabilitation Services Administration, and the National Federation of the Blind, are outlined. Preliminary tests of the machine with blind secondary school students have focused on the intelligibility of the synthetic speech output, and the results of these tests are presented.


The uptake kinetics and metabolism of extraneuronal noradrenaline in guinea-pig trachea as studied with quantitative fluorescence microphotometry. 1 Extraneuronal uptake of noradrenaline and alpha-methylnoradrenaline into single cells of guinea-pig tracheal smooth muscle have been studied by means of quantitative fluorescence microphotometry. 2 Fluorescence brightness due to accumulation of alpha-methylnoradrenaline was dose-dependent and was increased by the catechol-O-methyltransferase inhibitor drugs, tropolone or beta-thujaplicin (200 micrometer) but not by 3,4-dimethoxy-5-hydroxybenzoic acid (200 microgram). 3 Fluorescence brightness due to accumulation of noradrenaline was increased if animals were pretreated with the monoamine oxidase inhibitor drug, nialamide. 4 The study suggests that metabolism of amines by catechol-O-methyltransferase and monoamine oxidase can occur subsequent to extraneuronal uptake is guinea-pig tracheal smooth muscle. 5 The uptake of noradrenaline into tracheal smooth muscle was concentration-dependent, saturable, and had a Km of 156 micrometer.


Renal angiography problems in live kidney donors. We compared renal angiographic and operative findings in 88 live kidney donors. Fifty patients had normal arteriograms and uneventful nephrectomies. Abnormal renal parenchymal or main artery lesions were found in eight donors (9%). These included four renal artery stenoses; the kidney distal to the lesion was used for transplanting. Thirty one patients (35%) had multiple renal arteries. Only 24 of these (77%) were diagnosed at pre-operative angiography, though on review angiogram accuracy could be raised to 94%. Except for one patient the accessory renal arteries missed at angiography were tiny twigs; the small renal infarcts caused by ligating them did not impair transplant survival. There was one serious misdiagnosis of multiple renal arteries, preventable on review of the abdominal aortogram. We condiser  pre-operative angiography an essential step for the live kidney donor. It is the most accurate way of ensuring that his remaining kidney will be structurally sound. Finding multiple renal arteries has a decisive impact on management. Abdominal aortography rather than routine selective arteriography is the safe and therefore preferable approach to these patients.


Day care and short stay surgery for hernia. The duration of postoperative stay has been falling in recent years as more conditions are considered suitable for outpatient care. The introduction of short stay surgery not only relieves hospital waiting lists but also represents an economic advantage and has certain social benefits for the patient. The treatment of more patients as day cases reallocates the workload, putting increased responsibility on to outpatient clinics and general practitioner and community services. It also demands careful consideration of in-hospital operative and anaesthetic techniques to speed recovery and minimize complications.


Flow and function in machine-preserved kidneys. One hundred machine-preserved cadaver kidneys were transplanted irrespective of their flow rates on the preservation machine. Twenty-five per cent had flows of below 100 ml/min and 11 per cent had flows of below 80 ml/min. There was no correlation between the flow rate and function at 1, 3 or 12 months. There was no difference in the flow rate between those kidneys which functioned immediately and those which never functioned. A fall in flow rate was associated with acute tubular necrosis of longer duration but eventual function was not impaired. There was a higher incidence of early aggressive rejection in kidneys which functioned immediately. Preservation failure per se is a rare cause of primary non-function. Adherence to a policy of ignoring the flow characteristics of machine-preserved kidneys could make up to 25 per cent more kidneys available for transplantation.


Arterial anastomoses in renal transplantation. A consecutive series of 260 renal transplants was analysed to find the outcome of different arterial anastomoses. In 177 cases the renal vessel was anastomosed to the end of the internal iliac artery; 15 per cent developed vascular complications. Thirty-eight of these patients subsequently underwent allograft nephrectomy; none died from haemorrhage. In 83 patients the renal vessel was anastomosed end to side to the iliac artery, and vascular complications occurred in 14 per cent. Thirty-two patients subsequently underwent allograft nephrectomy; 8 were associated with severe haemorrhage and 5 of these were fatal.


The bacteriology of primary wound sepsis in potentially contaminated abdominal operations: the effect of irrigation, povidone-iodine and cephaloridine on the sepsis rate assessed in a clinical trial. Two hundred and nine potentially contaminated abdominal operations were randomly allocated to prophylaxis with a single dose of 1 g cephaloridine intraincisionally, irrigation of the wound at the end of the operation with saline or spraying of the wound with povidone-iodine. In high risk operations (ileocolorectal or those in obese patients) the rate of major wound sepsis in those protected by cephaloridine was 3.8% compared with 13.2% in the irrigation and 16.7% in the povidone-iodine groups. In low risk operations no significant differences in sepsis rates were found. Bacteriological studies of incised organs, subcutaneous fat and pus showed that the majority of wound infections arose from endogenous sources. The outstanding problem remains that of prevention of contamination of the abdominal wall during surgery.


Wound infection in emergency appendicectomy: a prospective trial with tropical ampicillin and antiseptic solution irrigation. A randomized controlled trial in 374 patients requiring emergency appendicectomy to determine the value of topical ampicillin and an antiseptic solution of chlorhexidine and cetyl trimethyl ammonium bromide (Savlon) in preventing wound infection is reported. The application of 1 g of ampicillin powder to the wound significantly reduced wound infection in perforated appendicitis, but not in unperforated appendicitis. Wound irrigation with 1% cetyl trimethyl ammonium bromide was ineffective in preventing wound infection in all grades of appendicitis. When wound contamination is difficult to avoid, as in perforated appendicitis, topical ampicillin should be used to reduce the rate of wound infection.


Cimetidine: prophylaxis against upper gastrointestinal haemorrhage after renal transplantation. The incidence of upper gastrointestinal haemorrhage within four months of renal transplantation was studied in two groups of patients. Thirty patients who received prophylactic cimetidine suffered no episodes of upper gastrointestinal haemorrhage, while six of the 33 patients who did not receive cimetidine suffered haemorrhages and one of them died as a result. The difference between the groups was significant. The results suggest that the prophylactic use of cimetidine in patients receiving renal transplants is worth while.


Antibodies to Candida albicans in human cervicovaginal secretions. The incidence of IgA and IgG antibodies against Candida albicans was determined in cervicovaginal secretions from 95 non-pregnant women. IgG antibodies were detected in 21.2% of women with vaginal candidosis, in 23.5% of women harbouring yeasts in the vagina without clinical signs of infection, and in 26.6% of women not harbouring yeasts in the vagina. IgA antibodies were found in 6.1% of women with vaginal candidosis, in 5.9% of women harbouring yeasts in the vagina without clinical signs of infection, and in 8.9% of women not harbouring yeasts in the vagina. IgG antibodies against C. albicans were detected in the serum of all 95 women. It is suggested that a proportion of the antibodies found in the secretions was derived from the circulation.


Immunohistochemical demonstration of S-100 protein and GFA protein in interstitial cells of rat pineal gland. The presence of the glial marker proteins, the S-100 and the glial fibrillary acidic (GFA) protein, in the pineal gland was investigated in the rat. Using both the indirect peroxidase-labelled immunoglobulin technique and the unlabelled antibody enzyme (PAP) method, we observed few scattered S-100 and GFA positive cells in the pineal. The number, location and morphology of these cells suggest they are the pineal interstitial cells. This indicates that the interstitial cells are of neuroectodermal origin, possibly macroglial cells themselves.


Cutaneous receptive field enlargement following skin-grafting in the frog, D. pictus. Misdirected wiping responses occur in Discoglossus pictus frogs when skin-grafted early in larval life. Cutaneous receptive fields (CRF) for various peripheral nerve branches were mapped electrophysiologically in control and skin-grafted frogs. The ventrolateral nerve branch (VL) frequently provided innervation for significant areas of dorsal skin in both control and skin-grafted animals. Although the VL crossed over onto dorsal skin areas in some skin-grafted frogs giving rise to misdirected reflexes, these dorsal extensions frequently failed to include all skin areas from which the maladaptive reflexes could be elicited. It was concluded that misdirected reflexes are mediated via dorsal nerve branches occupying normal mid-dorsal areas of the back.


Squamous cell carcinoma of the tongue in a nine year renal transplant survivor: a case report with a discussion of the risk of development of epithelial carcinomas in renal transplant survivors. A case of squamous cell carcinoma of the tongue in a 26-year-old man developing nine years after renal transplantation is presented. This is the first such case to be reported. Within seven months, widespread tumor metastases resulted in death. Pathologic examination of the transplanted kidney demonstrated neither re-establishment of glomerulonephritis nor evidence of rejection, and lack of significant renal disease was confirmed by electron microscopy. This long term survival without development of renal pathology is also of great interest.


Prednimustine (NSC-134087, Leo 1031) treatment of lymphocytic and lymphocytic-histiocytic lymphomas. Nineteen patients with advanced lymphocytic or lymphocytic-histiocytic lymphomas were treated with Prednimustine (NSC-134087, Leo 1031). The median induction dose was 25 mg/m2 a day by mouth (range 11-42). Ten patients had previously received radiation or chemotherapy, or both. Four patients had a complete remission and eleven a partial remission. The median duration of remission was 12.5+ months for complete responders and 5 months for partial responders. Thirteen patients had a moderate myelosuppression. One patient had urticaria and pruritus and refused further treatment.


Perspectives on the statistician's role in cooperative clinical research. Some contributions which the statistician can make to cooperative research are briefly discussed in the hope of furthering communication between statistician and clinician. In the planning stages the major issues are those of sample size, randomization, data collection and early termination. Careful consideration of these points can help ensure that the main questions posed by the trial are answered as economically as possible. During the final stages the major statistical contribution is in analysis and presentation of results in a way which explores the interrelationships between the most important baseline and therapeutic factors and their joint effect on outcome.


Primary malignant melanoma of the cerebellum: a case with metastases outside the nervous system. A case of primary intracranial melanoma in a 65-year-old female is described. A complete postmortem examination revealed multiple metastases in the liver, and eliminated the possibility of an extraneural primary melanoma. A review of the literature confirmed that such cases are highly exceptional, and that an exhaustive autopsy including particularly the eyes is indispensable to substantiate the diagnosis. When these conditions are fulfilled, there is no reason not to consider the eventuality of visceral deposits from a primary melanoma of the central nervous system.


The clinical pharmacology of methotrexate: new applications of an old drug. Methotrexate is now used widely for the treatment of acute leukemia, non-Hodgkin's lymphoma, osteogenic sarcoma, choriocarcinoma, breast carcinoma, pulmonary and epidermoid carcinoma, and intrathecal chemotherapy. It is also useful in bone marrow transplantation, severe psoriasis, rheumatoid arthritis, dermatomyositis, Wegener's granulomatosis and sarcoidosis. The recent dramatic intensification of methotrexate therapy can be attributed in part to advances in our understanding of the clinical pharmacology of the folate antagonists, as well as to the combination of positive results and their effective dissemination to medical oncologists. The review summarizes the pharmacologic findings and illustrates how they are currently being applied to the treatment of malignant disease.


Chronic granulocytic leukemia in a renal transplant recipient. Chronic granulocytic leukemia developed four years after renal transplantation in a patient, who had been treated with immunosuppressive agents since then. The low incidence of leukemia in comparison with malignant lymphoma in renal transplant recipients and some pathogenetic mechanisms are discussed.


Mutagenicity of some congeners of benzidine in the Salmonella typhimurium assay system. The analogs of benzidine were assay for mutagenicity using Salmonella typhimurium TA-98 and TA-100 and a mouse liver enzyme preparation. Only 4-aminobiphenyl produced both frameshift and base pair substitution mutations and 3,3'-dichlorobenzidine was the only compound which was mutagenic without the mammalian enzyme factor. When hydrochloride salts of the parent compounds were made to improve their stability for animal feeding experiments, the mutagenicity for the Salmonella tester strains was reduced except for 3,3'-dimethylbenzidine. Animal feeding trials in mice are underway to determine the dose response relationship of tumor incidence and molecular configuration.


The mutagenicity of airborne particulate pollutants. The mutagenic effect of extracts derived from airborne particulate matter was investigated with the Ames-test. The whole extract proved to be distinctly mutagenic. To find out whether the content of polycyclic aromatic hydrocarbons may be responsible for the mutagenic effect, the extract was split into several fractions. The fraction containing the polycyclic compounds showed the lowest mutagenic rate that was enzymatically mediated. The other fractions required metabolic activation for some of their components, but some components were active without metabolic activation.


Chromomycin A3 for advanced breast cancer: a Southwest Oncology Group study. Chromomycin A3 was administered iv to 26 patients in a phase I trial. The maximum tolerated dose established in this study was 0.75 mg/m2/day iv X 5 days. The drug was then given to 48 evaluable patients with far-advanced disseminated breast cancer. Two short partial remissions and one clinical improvement were seen. Toxic manifestations consisted of frequent and usually reversible renal toxicity, nausea and vomiting, occasional thrombocytopenia, hypocalcemia, and two instances of semicoma. Drug toxicity may have contributed to the death of two patients.


Current dilemmas in the management of non-seminomatous germ cell tumors of the testis. Testicular tumors are the second most common neoplasm in young adult males. Improved techniques of staging, surgery, radiation therapy and chemotherapy have markedly improved the survival of these patients during the past twenty years. However, the treatment of non-seminomatous germ cell tumors remains controversial. This paper reviews the current literature and suggests that methods of performing controlled clinical trials must be found if we are to learn the best methods of treating these patients.


Substructure of inner mitochondrial membranes of myocardial cells as shown by cryo-ultramicrotomy. The substructure of the inner mitochondrial membranes has been studied by cryo-ultramicrotomy under conditions during which denaturation of proteins by treatment with chemical solutes has been totally avoided. In such preparations, the inner membrane has a substructure consisting of globular subunits. These subunits have an average diameter of ca. 20 A-ca. 62 A and are fairly regularly spaced. Intracristal space is absent in the unstained, freeze-dried preparations, whereas a space of ca 40 A is seen in preparations lightly treated by OsO4-vapour. It is concluded that the subunits of the inner mitochondrial membranes probably consist either of single protein molecules or of complexes of protein molecules.


The distribution of LHRH in the hypothalamus of the thirsting rat. A light and electron microscopic immunocytochemical study. Prolonged thirst provokes an activation of the LHRH system which enables the visualization of fiber connections not seen in untreated control animals. This type of experimental stress situation increases the number of LHRH-containing fibers in the organum vasculosum laminae terminalis and in the median eminence. The number of LHRH-producing cells in the preoptic nucleus is increased and the fiber connection between this area and the median eminence can be observed. The tanycytes and the perikarya of the arcuate nucleus do not react with the antibody against LHRH. Moreover, during thirst, a network of LHRH-containing fibers is observed in the medial mammillary nucleus. The results obtained at the light microscopic level have been confirmed and supplemented by electron microscopic immunocytochemical observations.


Localisation of somatostatin- and gastrin-like immunoreactivity in the gastrointestinal tract of Ciona intestinalis L. Somatostatin- and gastrin-like immunoreactivity has been found by immunofluorescence in cells of the stomach and intestinal epithelia of Ciona intestinalis L. The cells containing the peptide immunoreactive to mammalian anti-gastrin can be restained with Grimelius' technique for argyrophilia.


The effect of the ionophore A23187 on amylase release, cellular integrity and ultrastructure of mouse pancreatic acini. The effects of the Ca2 + ionophore A2317 on pancreatic amylase and lactate dehydrogenase (LDH) release, cellular electrolyte balance and ultrastructure were studied with the use of incubated pancreatic fragments. A23187 (0.3 micrometer) in the presence of Ca2 +, increased amylase release but at higher concentrations (1-10 micrometer) also increased LDH release and increased uptake of 14C-sucrose with concomitant loss of tissue K + and gain in Na +. The ultrastructure of the majority of acini appeared normal and showed depletion of zymogen granules. Microtubules and microfilaments which have been implicated in the release process were normal or increased in number. In the absence of Ca + the ionophore had no effect on secretion, cellular integrity or ultrastructure. It is concluded that A23187 in the presence of Ca2 + increases amylase release by a mechanism comparable to the terminal steps in stimulus-secretion coupling induced by physiological secretagogues. This provides further evidence that amylase release is mediated by a rise in cell Ca2 + although the mechanisms of the ionophore- and physiological secretagogue-induced rise in Ca + are probably different. High concentrations of ionophore (greater than 1 micrometer) also induce Ca2 + dependent damage in a fraction of the cells.


beta-Lipotropin in brain: localization in hypothalamic neurons by immunoperoxidase technique. beta-Lipotropin (beta-LPH) has been localized in hypothalamus and pituitary of sheep and ox by the immunoperoxidase technique. In both species beta-LPH was found in perikarya of arcuate neurons as well as in cells of the anterior and intermediate lobes of the pituitary. A large number of immunoreactive axons were found in the arcuate region; some appeared to innervate other neurons and others projected to portal capillaries. Stained fiber segments were also scattered throughout the hypothalamus. The presence of beta-LPH in hypothalamic neurons supports the possibility that brain beta-LPH may be a precursor for opiate-like or other peptides which may be involved in neuromodulation or neurohormonal activities.


Intra- and extrahypothalamic vasopressin and oxytocin pathways in the rat. Perfusion of rat brain followed by immersion fixation with 2.5% glutaraldehyde-1% paraformaldehyde, purification of the first antisera and application of the unlabelled antibody enzyme method were used to specifically identify vasopressin and oxytocin containing cells and fibres. The conventional sites of production of these hormones were confirmed as follows: supraoptic and paraventricular nuclei, suprachiasmatic nucleus (only vasopressin), and other cells and cell groups of the hypothalamus. Fibres from the suprachiasmatic nucleus spread out in various direction, and probably project to the nucleus praeopticus periventricularis, organum vasculosum laminae terminalis and in the direction of the supraoptic nucleus. Oxytocin and vasopressin containing pathways could be traced from the paraventricular nucleus to the lateral ventricle, the stria terminalis and the stria medullaris. Some of the oxytocin and vasopressin containing tracts appear to continue onto ther septum. The possible importance of these morphological findings for the behavioural effects of vasopressin and oxytocin is discussed.


Immunocytochemical demonstration of calcitonin-containing C-cells in the thyroid glands of different mammals. In the thyroid glands of the horse, pig, deer, mole, and rat, C-cells could be demonstrated by means of the immunocytochemical PAP-technique using rabbit antisera against human calcitonin. Only in ruminants, the cross-reaction between the intracellularly stored antigen and the antibodies used appeared to be incomplete.


Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. An antiserum has been found in a nonimmunized rabbit which reacts strongly with a system of filaments in various fibroblasts, epithelial cells, macrophages and neuroblastoma. These filaments are distinct from the actin microfilament bundles visualized by an antibody against actin, and they are not affected by brief treatment with cytochalasin B. The pattern of these filaments somewhat resembles that described for microtubules, but the filaments could be clearly distinguished from microtubules by a comparison of their respective immunofluorescent patterns during cell division. In response to the drugs colcemid and vinblastine, the filaments reacting with this preimmune serum condense to form a compact perinuclear coil of fibers, a distribution and behavior in agreement with that previously described for the 10 nm or intermediate filaments studied by electron microscopy. Further evidence supporting our conclusion that this antiserum reacts with intermediate filaments is provided by a comparison of electron micrographs and the immunofluorescent patterns from parallel cell cultures. To identify the antigens reacting with this antiserum we have used the new technique of immuno-autoradiography on SDS gels of whole cell extracts. Two reactive polypeptide chains have been identified with apparent molecular weights of 56,000 and 30,000 daltons.


Some rRNA operons in E. coli have tRNA genes at their distal ends. We have previously isolated seven rRNA operons on plasmids or lambda transducing phages and identified various tRNAs encoded by these operons. Each of the seven operons has one of two different spacer tRNA gene arrangements between the genes for 16S and 23S rRNA: either tRNAGlu2 or both tRNAIle1 and tRNAAla1B genes. In addition, various tRNA genes are located at or near the distal ends of rRNA operons. In particular, genes for tRNATrp and tRNAAsp1 are located at the distal end of rrnC at 83 min on the E. coli chromosome. Experiments with various hybrid plasmids, some of which lack the rRNA promoter, have now demonstrated that this promoter is necessary for expression of the distal tRNA genes. Rifampicin run-out experiments have also provided evidence that the tRNATrp gene is located farther from its promoter than the spacer tRNA gene or the 5S RNA gene. These results confirm the localization of genes for tRNATrp and tRNAAsp1 at the distal end of rrnC and strongly suggest that they are co-transcribed with the genes for 16S, tRNAGlu2, 23S and 5S RNA. Other such distal tRNAs have been identified, and it is suggested that they too are part of rRNA operons.


The first decade of aortocoronary bypass grafting, 1967-1977. A review. Despite a decade of experience with aortocoronary bypass grafting embracing 300,000 or more operations, indications for its use remain controversial. The controversy persists because of a lack of adequate controls with which to compare the clinical course of operated patients; only 1248 have been reported who have been studied in a carefully controlled and random manner. Benefit has been claimed frequently by comparing the course of patients treated surgically with medically treated patients followed the decade before. Such comparisons are not valid in view of the well documented changes in the natural history of coronary artery disease that have been occurring during the last decade. Despite a low operative mortality and rate of graft closure, available data in the literature do not indicate that initial symptomatic improvement necessarily persists, or that myocardial infarctions, arrhythmias, or congestive heart failure will be prevented, or that life will be prolonged in the vast majority of operated patients.


Use of a programmable calculator in processing and interpreting serum cholinesterase phenotypes. Cholinesterase phenotyping is done at 25 degrees C with use of benzoylcholine as substrate and dibucaine, fluoride, chloride, and succinyldicholine as inhibitors. We wrote a diagnostic program in modified BASIC language for the processing and interpretation of these cholinesterase phenotypes. We used the HP 9830A calculator. The diagnostic aspect of the program uses 646 words, a further 86 words being used to run this program. The traditional and the programmed reporting procedures were duplicated for 296 consecutive patients, with no case of disagreement between the two reporting methods as to the appropriate phenotype. The programmed system requested nine repeat analyses more than did the traditional method. Seven of these were due to the tight limits set in the program for the fluoride inhibitor numbers. These tight limits were set to ensure that there was as limited an overlap as possible between the phenotypes E1uE1u and E1uE1f. The remaining two repeats were due to the restrictions placed on the succinyldicholine inhibitor numbers that would be acceptable before a patient was designated as having the relatively rare phenotype, E1aE1f.


Closure of anterior palate fistulae.

Closure of anterior palate fistulae. Attempts at closure of anterior palate fistulas using local tissue have resulted in a high rate of failure. In addition most of these patients have associated maxillary collapse which must be corrected prior to surgery. A technique using a buccal mucosal flap to gain unscarred tissue for the anterior closure and bone grafts to fill the bony defect has been performed in nineteen patients with anterior palate fistulas. The results have been satisfactory from a functional and esthetic standpoint. In addition associated deformities have been corrected simultaneously.


A comparison of two creatine kinase MB determinations: a chromatographic versus an immunological method. Determinations of the MB isoenzyme of creatine kinase by ion-exchange chromatography and by an immunological method show good overall correlation in plasma of patients with proven infarctions, r=0.98. In analyzing myocardial tissue samples, unexpected discrepancies are found between these two methods. An as-yet-unidentified creatine kinase, in chromatography behaving as MM and immunologically undistinguishable from MB, is found.


Treatment of senile osteoporosis with 1alpha-hydroxyvitamin D3. About 30% of patients with clinical osteoporosis had histological signs of osteomalacia, in spite of normal serum 25-hydroxyvitamin D3 (25-OHD3). The excess osteoid disappeared during treatment with 1alpha-hydroxyvitamin D3 (1alpha-OHD3). These patients might have reduced ability to convert 25-OHD3 to 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3). The intestinal calcium absorption increased during treatment with 1alpha-OHD3, but this was accompanied by a rise in urinary calcium excretion. Photon absorptiometry of the forearm indicated increased bone mineral content during treatment with a daily dose of 2 microgram 1alpha-OHD3 and a supplement of 1 g of calcium. This therapeutic combination, however, caused frequent episodes of hypercalcaemia, so further studies are necessary to evaluate an appropriate dose of 1alpha-OHD3 with or without a calcium supplement.


Physiological and therapeutic differences between vitamin D, its metabolites and analogues. Since several metabolites and analogues of vitamin D have become available, their physiological and therapeutic potential have been studied in various disorders of mineral metabolism, particularly those classically termed vitamin D-resistant. In some instances this resistance appears to be due to defective metabolism of vitamin D or to defective target organ responses, but in others it is not. Although the precise effect of vitamin D or its metabolites on mineralization of bone is still unknown there is some evidence that the metabolites differ in their relative effects on intestine, bone, muscle and parathyroid glands. These differences may eventually have therapeutic value. At present the major advantage of 1alpha-hydroxylated derivatives of vitamin D over other compounds resides in their more rapid onset and reversal of action which makes them much easier and safer to use.


Diagnostic procedures in disorders of calcium metabolism. The investigation of a patient with a suspected disorder of calcium metabolism is most rapidly and efficiently performed by observing a standard procedure, the initial stages of which are simple and mandatory, the later stages more complex and determined by the initial results and the depth of investigation required. Needless to say, the investigations are preceded by history taking and routine physical examination. The following account is a summary of available procedures, most of which have been described elsewhere in greater detail (Nordin et al., 1976a).


Circulating immune complexes in six patients with trichinosis. The sera of six patients infected with Trichinella spiralis were studied for the presence of circulating immune complexes (CIC). CIC were present in all six patients studied 1 month after infection. In two patients in whom serial serum samples were available, as clinical improvement occurred there was a decrease in the level of CIC and an increase in the fluorescent antibody titres.


Immunological studies of human placentae: the distribution and character of immunoglobulins in chorionic villi. All four human IgG subclasses, and both kappa and lambda light chains, were detected by immunofluorescence in similar distributions in chorionic villi of human placentae. IgG1 and IgG3 were the predominant subclasses. No evidence was obtained for local enzymatic digestion of IgG during placental transfer. Most of the IgG on the trophoblastic basement membrane (TBM) was loosely bound and could be removed by prolonged washing, although some appeared to be more tightly bound to small segments of the TBM. IgM, but not IgA, was present in small amounts in placental villous structures. Immunoglobulin was never observed within the syncytiotrophoblast. Antisera to IgG genetic (Gm) markers were used to locate IgG thought to be of foetal or maternal origin. The presence of paternal Gm markers not carried by the mother was taken as evidence for foetal IgG. Foetal (paternal) Gm markers were observed in placentae, although maternal IgG was the major immunoglobulin present in placental villi. Both maternal and foetal IgG were demonstrated in fibrinoid deposits, vessel walls and the cytoplasm of some stromal cells. Only foetal IgG was definitively observed in the immunoglobulin that is tightly bound to the TBM.


Interstitial nephritis in rats produced by E. coli in adjuvant: immunological findings. An increased incidence and severity of interstitial nephritis was produced in F344/fmai rats immunized with E. coli 022 in pertussis vaccine for 12-15 months. Migration of peritoneal exudate cells from immunized animals was inhibited by syngeneic kidney antigens. One out of twenty-eight immunized animals developed anti-TBM antibodies. In this model, interstitial nephritis develops in association with cell-mediated immunity to kidney tissue.


Deficiency of serum bactericidal activity against Salmonella typhimurium in sickle cell anaemia. Systemic salmonellosis is a recognized complication of sickle cell anemia (SCA). In our initial study of SCA host defences against salmonella, we evaluated the bactericidal activity of serum against Salmonella typhimurium. When compared to controls, sera from eight out of nineteen SCA patients were deficient in bactericidal function. Levels of factor B, haemolytic complement and agglutinating antibody were similar in SCA and control sera. However, abnormalities that might theoretically account for the decreased antibacterial activity were observed in many SCA sera. These abnormal findings included: (a) defective function of the alternative complement pathway (decreased bacterial killing in the presence of Mg EGTA); (b) low serum C3 concentration; and (c) decreased total iron-binding capacity (TIBC), with a resultant increase in per cent saturation of iron-binding capacity. Of these deficiencies only the abnormal alternative pathway function was significantly associated with decreased serum bactericidal activity. A suggested function of serum bactericidal activity is prevention of bacteraemia by susceptible organisms. Thus diminished serum bactericidal capacity may increase the risk of Salmonella bacteraemia in some individuals with sickle cell disease.


Cell-mediated immunological reactivity in neonates and infants with congenital syphilis. Thirty-eight mothers and babies were assessed by the lymphocyte transformation test for their cell-mediated immune responses to Treponema pallidum and to purified protein derivative of tuberculin (PPD). All babies, including controls, appeared to be reactive when results of [3H]-thymidine incorporation were expressed as net counts per minute. This was because of high background activity, which was most marked in cells from neonates. When results were expressed as stimulation ratios, it was revealed that cells from neonates were unreactive to both antigens, even when those from their mothers were reactive. Cells from some older babies with active congenital syphilis were reactive to both antigens.


Characterization of cell subpopulations of the thymus by a hydrophobic fluorescent probe: 1-anilino-8-naphthalene sulphonate. 1-anilino-8-naphthalene sulphonate (ANS) is a hydrophobic fluorescent probe. ANS is not fluorescent in water or in RPMI 1640 medium, but it becomes fluorescent when bound to hydrophobic components of cells. The binding of ANS to mouse thymocytes shows that two cell subpopulations are identified, which differ in their emission wave-lengths, as reflected by their distinct 'blue' and 'green' fluorescence. This difference correlates with the DNA content of cells and this technique may characterize thymic cells in the G1 or in the S-G2 phases of the cell cycle in a quick and simple way.


The effect of cyclophosphamide on an ocular immune response. I. Primary response. The ocular inflammation and antibody production that follow intravitreal injection of rabbit eyes with bovine gamma globulin were suppressed by treatment with Cytoxan by the intramuscular (i.m.) route. The drug suppressed PFC responses of uveal tract and corneal cells when it was administered, beginning as late as 5 days after immunization, if treatment was continued until day 12 or 13. Short-term treatments and treatment with smaller Cytoxan doses were less effective. We noted a good correlation between the presence or absence of ocular inflammation, suppression of ocular PFC responses and depression of serum, aqueous humour and vitreous humour antibody titres. In some treatment groups ocular antibody production seemed to be completely suppressed, while in others antibody production was significantly delayed.


Studies on the heterophile antibodies of infectious mononucleosis. I. Separation of four antibody populations, one of which contains lymphocytotoxic activity. With the aid of insolubilized immunoadsorbents made with glycoproteins, extracted from cow red blood cells (RBC), and with guinea-pig kidney tissue, heterophile antibody populations of the Paul--Bunnell type and the Forssman type were eluted from infectious mononucleosis sera. Two Paul--Bunnell-type antibody populations were separated on the basis of their affinity for cow RBC antigens. The lymphocytotoxic antibodies in the infectious mononucleosis sera were eluted among the low-affinity Paul--Bunnell antibodies.


Multiple myeloma and monoclonal IgA with anti-actin reactivity. Serum containing a monoclonal IgA protein from a patient with multiple myeloma gave intense immunofluorescent staining of smooth muscle fibres and the striations of skeletal muscle, cardiac muscle and thymic myoid cells. It also gave a weaker reaction with hepatocytes in a 'polygonal' pattern, and with renal glomeruli in a diffuse pattern. In culture fibroblasts, the serum stained long, parallel cytoplasmic filaments. Specificity of the staining reactions for actin was established by their prevention on serum absorption with skeletal muscle actin, but not by skeletal muscle myosin, tropomyosin or troponin, and by the demonstration that eluates obtained by acid dissociation of the serum-actin precipitates gave the same staining reactions as the original serum. Localization of the anti-actin reactivity to the monoclonal IgA protein was established by the observation that the same staining reactions were obtained with a monospecific IgA conjugate, with the eluate derived from the gamma globulin band of a serumel ectrophoretic strip, and with a purified euglobulin (IgA) fraction; also, the eluates obtained by acid dissociation of the serum actin precipitates contained monoclonal IgA.


A study of maternal lymphoid organs and the progeny following treatment with immunomodulating agents during pregnancy. Whether differences in foetoplacental weight and post-implantation mortality in rodents are secondary to heterosis and inbreeding depression or antigenic differences between mother and foetus has been a continuing controversy. To determine whether non-specific depression or stimulation of the maternal immune system affects the success of the foetoplacental allograft, groups of virgin Fischer (Ag-B1) females of similar age and weight mated with DA (Ag-B4) males were treated with daily intraperitoneal injections of: (a) saline, (b) methylprednisolone (MP), 1-0 mg/kg, (c) cyclophosphamide (CY), 3.0 mg/kg, or (d) azathioprine (AZ), 3.0 mg/kg; or they were injected intraperitoneally on the fifth day of gestation with: (a) B. pertussis, 1.0 ml, (b) C. parvum, 0.2 ml, or (c) BCG, 0.1 ml. None of the immunostimulating agents were detrimental to the progeny, but the immunosupprissive drugs caused an increased percentage of foetal deaths and foetoplacental growth retardation. The reduced foetal and placental size induced by CY or AZ could be partially blocked by simultaneous maternal treatment with BCG. Analysis of mean maternal weight gain, spleen weight assays, changes in the lymph nodes draining the uterus and comparison of data from non-pregnant animals and syngeneic pregnancies treated with these agents suggest that immunosuppressive drugs reduce foetal survival rates and produce foetoplacental growth retardation via a combination of immunological and cytotoxic mechanisms.


Response of asthmatic patients to fenoterol inhalation: a method of quantifying the airway bronchodilator dose. A radiotracer technique is described which enables direct measurement of the dose and distribution of inhaled aerosol bronchodilator in man. The mean (+/-SD) amounts of the B2-adrenergic agonist, fenoterol, administered to a group of 12 asthmatic subjects in a double-blind randomized fashion were: placebo, 0 microgram; low dose, 5.6 (+/-1.2) microgram; medium dose, 32.7 (+/-7.3) microgram; and high dose, 127.5 (+/-29.2) microgram, with a mean of 86.3% of the total subject dose being deposited in the lungs. The medium and high doses of fenoterol produced similar increases above baseline in forced expired volume in 1 sec (FEV1), maximum flow at 50% of vital capacity (V max 50), and maximum flow at 25% of vital capacity (V max 25). These increases were greater than those with placebo for the entire 4-hr study (p less than 0.01). The low dose of fenoterol was more effective than placebo in increasing FEV1, V max 50, and V max 25 above baseline values (p less than 0.05), but not for the entire 4-hr study. The high-dose fenoterol caused palpitations and tremor in 3 of the 12 subjects, and the medium-dose fenoterol caused palpitations in one of these subjects.


Current treatment of acute leukemia.

Current treatment of acute leukemia. For patients with ALL we must be able to identify clearly both standard and high-risk patients. For standard-risk patients in whom life expectancy is long, therapy regimens must be designed to provide optimal control with minimal short- and long-term toxicity. For patients with high-risk ALL, methods must be found to circumvent the problem of early acquisition of drug resistance by the leukemic lymphoblasts. For patients with AML, more work must be done to improve the results of remission induction. But the major problem for these patients is the maintenance of a remission once it is obtained. Perhaps one of the critical factors in AML is the interrelationship of normal and leukemic marrow elements in therapy response. Disease-free survival time must be improved before considerations of CNS prophylaxis and cessation of therapy can be approached.


Caries-preventive effect of two concentrations of stannous fluoride mouthrinse. The effectiveness of a stannous fluoride mouthrinse, when used once each school day, was investigated in a 3-year study. Effervescent stannous fluoride tablets of two concentrations were dissolved in 20 ml of water, giving solutions of 100 parts/10(6)F- and 200 parts/10(6)F- respectively. Approximately 1,200 children, with a mean age of 10 years, were divided on a random basis into three groups. Two of the groups rinsed with the two strengths of solution and the third group rinsed with a placebo. Examinations were carried out at the commencement of the study, and at yearly intervals thereafter. The final series was carried out a year after the rinsing procedures were terminated. There were significant reductions in the numbers of new caries in each of the two experimental groups as compared with the controls. The concentration of the solutions appeared to have little influence on the results. More dramatic reductions were noted in the teeth which erupted during the course of the study. A residual effect was demonstrated a year after the rinsing procedures were terminated.


Comparison of blind and non-blind assessments of occlusal sealant retention. A comparison was made of scoring the retention status of BIS-GMA occlusal sealants 5 years after they had been placed, according to a non-blind and blind method of identification of teeth that had been sealed. Subjects were 175 children ages 15--18. One examiner made all the assessments. Retention of sealant was classified as all present, partly missing or all missing. There was 90.5% complete agreement between non-blind and blind examinations in scoring the retention status of 893 occlusal sites. Essentially no difference in agreement was found between sites on maxillary and mandibular teeth nor according to sites in specific teeth. There was only a slight tendency to classify more sites as having sealant present at the non-blind examinations than at the blind examinations, 56.7% and 55.1%, respectively. These findings would seem to indicate that whether examinations are done blindly or non-blindly is of minor importance in making an accurate assessment of sealant retention.


Adequte cohort sizes for caries clinical trials. Methods of determining cohort sizes were examined to determine their appropriateness for use in multi-group caries clinical trials. The appropriate method to sue depends on the type of trial being planned. It is shown that multiple comparison methods using certain Bonferonni type t-statistics ought to be used in trials in which different levels of frequencies of application of known caries inhibitors are being tested. It is also demonstrated that using the F-test procedure in the determination of cohort sizes can result in unacceptable low sensitivity levels being realized for the comparisons of primary concern. Tables are presented which can be used to determine group sizes needed to achieve specified sensitivity levels for two-groups trials and multi-group trials.


Toxins produced by Candida albicans.

Toxins produced by Candida albicans. From studies on high-and-low molecular-weight candidial toxins, with special reference to the etiopathology of experimental-weight candidial toxins, with special reference to the etiopathology of experimental C. albicans infection, it may be concluded that these toxins, particularly the high-molecular-weight ones, play a very important role as parasitic factors in the extablishment of the infection, possibly also affecting its course. This concept of the role of Candidial toxins may be considered also applicable to the establishment of natural candidosis. It is speculated that these toxins might be more or less ubiquitously produced in pathogenic fungi, not only C. albicans but also other species of the genus Candida as well as those of other fungal genera. They may be to some extent related to the etiopathology of their respective fungal infections.


Cryptococcosis: a diagnostic challenge.

Cryptococcosis: a diagnostic challenge. Human cryptococcosis is an exogenous human infection of increasing frequency and growing importance. Early diagnosis is essential for the clinical outcome, no matter whether the infection is primary or secondary to underlying disease. Reliable and quick diagnostic methods are outlined and some pitfalls are discussed. Direct microscopy is essential, followed by cultivation procedures and serologic tests.


A comparison of aphasic and normal adults in a sentence-verification task. This study showed that in certain sentence-processing tasks, aphasics fail to perform correctly because they fail to execute particular mental operations. A straightforward information-processing model describes the mental processes that are used in deciding whether a sentence is true or false of an accompanying picture. The aphasics' performance, interpreted in light of the model, indicates what step in their processing is not operating properly in this task.


Decreased serum complement in the Gardner-Diamond syndrome: immunofluorescent findings and association with angioimmunoblastic lymphadenopathy. Immunologic and immunofluorescent profiles of two patients with the Gardner-Diamond syndrome are described. During the time ecchymoses were present, both patients had decreased serum complement levels; when the lesions healed, the serum complement level returned to normal. One patient had associated angioimmunoblastic lymphadenopathy and subepidermal deposits of immunoglobulin IgM at the basement membrane of ecchymotic skin. The other patient had a normal immunofluorescent pattern. Both patients had increased B-cell counts. The association of immunologic and immunofluorescent findings in patients with the Gardner-Diamond syndrome is suggestive and deserves further study.


[In-patient treatment of peptic ulcer with cimetidine. I. Effect on duodenal ulcer healing (author's transl)]. The effect of 300 mg cimetidine q.i.d. on ulcer healing was studied in a controlled double-blind clinical trial of 71 in-patients with duodenal ulcer. Healing occurred in 48.5% of patients in the cimetidine group after two weeks, and in 20.6% in the placebo group (P less than 0.05). The healing rate was 88% in the cimetidine group at four weeks, 79.4% in the placebo group. Only during the first day was ulcer pain significantly reduced in the cimetidine-treated patients. Neither basal nor pentagastrin-stimulated acid and pepsin secretions were affected by 17-day administration of cimetidine. The drug had to be withdrawn in two patients because of elevated serum-creatinine levels. There was no other untoward effect.


[In-patient treatment of peptic ulcer with cimetidine. II. Controlled double-blind trial on gastric ulcer patients (author's transl)]. In a controlled double-blind clinical trial of 39 in-patients with gastric ulcer the effect of cimetidine on ulcer healing, ulcer pain and pentagastrin-stimulated acid and pepsin secretion was measured. A faster healing rate in the cimetidine group was statistically not significant. Cimetidine had no effect on ulcer pain and pentagastrin-stimulated acid and pepsin secretion. There were no serious untoward reactions.


[Prolonged renal graft survival induced by blood transfusion (author's transl)]. All renal allotransplants performed at the Cantonal Hospital, Zurich, in 1975 and 1976 were analysed with respect to pre-transplant blood transfusions, excluding secondary transplants, patients on dialysis outside of Switzerland, combined renal and pancreatic transplants, and women with previous pregnancies but without transfusions. Among 72 patients left for analysis there were 16 who had never received any pre-transplant blood transfusion. Their graft survival was much worse (one-year graft survival 36%) than that in patients with previous transfusions (one-year graft survival 70--80%). Patients without previous blood transfusion should, therefore, no longer be admitted for renal transplantation.


Fenoterol: a review of its pharmacological properties and therapeutic efficacy in asthma. Fenoterol, the 4-hydroxyphenyl derivative of orciprenaline, is a resorcinol derivative with relatively high selectivity for beta2-adrenoceptors. It is active in man after inhalation or oral administration and is indicated in the treatment of bronchospasm associated with asthma, bronchitis and other obstructive airway diseases. Clinical experience has shown that fenoterol is an effective bronchodilator with negligible effects on the cardiovascular system following aerosol administration of usual therapeutic doses. In children, inhaled fenoterol is effective in preventing exercise-induced asthma and administration of the aerosol in young children has been successfully used to terminate acute asthma attacks. In trials in adults, inhaled fenoterol was superior to placebo. In other controlled studies, it showed a tendency to cause a slightly greater maximum improvement in airway function as assessed spirometrically, and to have a longer duration of action than inhaled orciprenaline, salbutamol or terbutaline, although in these trials statistically significant differences were often not found. The onset of maximum effect is less rapid than with isoprenaline but is longer lasting. About 60% of the eventual maximum response to fenoterol is reached in the first few minutes after inhalation. Oral fenoterol is more effective than placebo, ephedrine or orciprenaline, and probably similar to salbutamol and terbutaline. Following usual aerosol doses, side-effects are minimal. Oral administration is associated with a higher incidence of side-effects than inhalation, including fine muscle tremor and tachycardia.


Loperamide: a review of its pharmacological properties and therapeutic efficacy in diarrhoea. Loperamide, a butyramide derivative is a new agent for use in symptomatic control of acute non-specific diarrhoea and chronic diarrhoea. Unlike diphenoxylate or codeine, loperamide does not appear to exert opiate activity in man at normal therapeutic doses. In acute diarrhoea, loperamide provides more rapid control of symptoms than diphenoxylate when given in a flexible dosage according to unformed bowel movements, and in single dose studies 4mg loperamide has a much longer duration of effect than 5mg diphenoxylate. Loperamide is probably superior to diphenoxylate in providing symptomatic control of chronic diarrhoea such as that associated with chronic inflammatory bowel disease or following gastrointestinal surgery. It has been used for up to 3 years in such conditions without evidence of tolerance. The possibility of once daily dosage of loperamide in chronic diarrhoea is an advantage. Side-effects have not proved a problem.


Drug-induced agranulocytosis.

Drug-induced agranulocytosis. An unexpected precipitous fall in peripheral leucocyte count may occur during treatment of certain sensitised individuals with drugs usually well tolerated by most people. Three basic mechanisms for drug sensitivity have been found. One is characterised by sudden destruction of large numbers of leucocytes in peripheral blood by antibodies elicited in response to drug sensitivity. A prototype for this type of reaction is aminopyrine. A second mechanism involves the production of a lupus-like syndrome followed by leucopenia in response to sensitisation to drugs such as procainamide. A third type involves development of agranulocytosis following a latent period during which a sensitive patient is treated with large amounts of chlorpromazine. This type of reaction is associated with production of bone marrow insufficiency in a patient who is believed to have a limited proliferative potential of bone marrow cells, which limit compensatory bone marrow response during treatment with a drug (e.g. chlorpromazine) that has limited bone marrow toxicity.


Cimetidine: a review of its pharmacological properties and therapeutic efficacy in peptic ulcer disease. Cimetidine is a specific competitive histamine H2-receptor antagonist which effectively inhibits gastric acid secretion and is advocated for the treatment of chronic peptic ulceration, haemorrhage from erosive gastritis, and the control of gastric hypersecretion and peptic ulceration in the Zollinger-Ellison syndrome. Placebo-controlled trials in outpatients have demonstrated its efficacy in promoting the healing of endoscopically diagnosed duodenal ulceration, during a period of 4 to 6 weeks, but its role in the treatment of gastric ulcer is less clear. Preliminary evidence suggests that maintenance therapy with cimetidine reduces the rate of recurrence of duodenal ulcer, but further studies are required to clarify its role in this situation and in the treatment of oesophagitis and acute gastrointestinal haemorrhage. Cimetidine controls the peptic ulceration of Zollinger-Ellison syndrome in most patients when given continuously for up to 2 years. Side-effects have generally been trivial and have very seldom necessitated withdrawal of therapy except in the rare occurrence of gynaecomastia. The haematological abnormalities particularly agranulocytosis, which lead to the withdrawal from clinical use of metiamide, have not been reported with cimetidine, except for 1 case of transient neutropenia. The safety of long-term cimetidine administration has yet to be determined.


Properties of the main endonuclease specific for apurinic sites of Escherichia coli (endonuclease VI). Mechanism of apurinic site excision from DNA. The main endonuclease for apurinic sites of Escherichia coli (endonuclease VI) has no action on normal strands, either in double-stranded or single-stranded DNA, or on alkylated sites. The enzyme has an optimum pH at 8.5, is inhibited by EDTA and needs Mg2+ for its activity; it has a half-life of 7 min at 40 degrees C. A purified preparation of endonuclease VI, free of endonuclease II activity, contained exonuclease III; the two activities (endonuclease VI and exonuclease III) copurified and were inactivated with the same half-lives at 40 degrees C. Endonuclease VI cuts the DNA strands on the 5' side of the apurinic sites giving a 3'-OH and a 5'-phosphate, and exonuclease III, working afterwards, leaves the apurinic site in the DNA molecule; this apurinic site can subsequently be removed by DNA polymerase I. The details of the excision of apurinic sites in vitro from DNA by endonuclease VI/exonuclease III, DNA polymerase I and ligase, are described; it is suggested that exonuclease III works as an antiligase to facilitate the DNA repair.


Immunochemical measurement of conformational heterogeneity of poly(inosinic acid). Several pure poly(I) preparations differed in: (a) their complement fixation reactivity with anti-poly(I) antiserum; (b) their ability to bind to a solid-phase anti-poly(I) antibody-Sepharose column; (c) their ability to inactivate serum complement; and (d) their reactivity with purified antibodies to double-stranded RNA. In particular, poly(I) samples that could induce interferon production differed from non-inducer poly(I)s; the inducers reacted weakly with anti-poly(I) antiserum and were the only ones that reacted with antibodies to double-stranded RNA. One inducer poly(I) did not inactivate complement, and differed from non-inducer poly(I) in quantitative aspects of poly(I) . poly(C) formation with varying amounts of poly(C). An additional type of poly(I) preparation reacted poorly with anti-poly(I) antiserum, did not react with anti-double-stranded-RNA antibodies and failed to induce interferon production. The varying forms of poly(I) were not interconvertible by boiling and rapid chilling. These results indicate that several different stable structural forms of poly(I) may result from a standardized synthetic procedure.


Blue-dextran--Sepharose affinity chromatography: recognition of a polynucleotide binding site of a protein. Native Escherichia coli polynucleotide phosphorylase can be retained on blue-dextran--Sepharose. The bound enzyme cannot be displaced by its mononucleotide substrates such as ADP, UDP, CDP, GDP and IDP, but it is easily eluted by its polymeric substrates. Under identical conditions, lactate dehydrogenase, bound on blue-dextran--Sepharose, is not eluted by poly(I) but can be specifically displaced by NADH. On the other hand, the trypsinized polynucleotide phosphorylase, known to be an active enzyme which has lost its polynucleotide site, does not bind to the affinity column. The native polynucleotide phosphorylase can also be tightly bound to poly(U)--agarose and displaced from it only by high salt concentration. The trypsinized enzyme is not bound at all on poly(I)--AGAROSe. Moreover, the native enzyme linked on blue-dextran--Sepharose, remains active indicating a free access of nucleoside diphosphates to the active center. These results taken together show that the dye ligand is not inserted onto the mononucleotide binding site and suggest rather that it binds to the polynucleotide binding region. The implications of this study and the application of blue-dextran--Sepharose affinity chromatography to other proteins having affinity for nucleic acids are discussed.


Alterations in tRNAs containing 2-methylthio-N6-(delta2-isopentenyl)-adenosine during growth of enteropathogenic Escherichia coli in the presence of iron-binding proteins. Escherichia coli grown in chemically produced iron-deficient media have well characterized alterations in the chromatographic properties of tRNAs containing the modified nucleoside 2-methylthio-N6-(delta2-isopentenyl) adenosine. The present report shows that similar tRNA alterations occur in enteropathogenic E. coli inhibited by human milk and bovine colostrum, the inhibited bacteria containing 10% or less of the normal tRNA species. Adding sufficient iron to saturate the iron-binding capacity of the lactoferrin present in milk and colostrum reversed these changes which are probably due to a failure to methylthiolate the isopentenyladenosine. Although adding iron led to a rapid replacement of abnormal tRNA by the chromatographically normal species, and to a resumption of multiplication, the tRNA alterations are not directly related to the inhibition of growth. Strains of E. coli which grew normally in milk, colostrum and in defined media containing the iron-binding protein transferrin or ovotransferrin also contained about 90% of the abnormal species. Rapid conversion of abnormal tRNA to normal tRNA occurred on adding iron and in the absence of RNA synthesis. The tRNA changes are discussed in relation to their possible connection with both the adaptation of E. coli to growth under the iron-restricted conditions imposed by iron-binding proteins in tissue fluids and with bacterial pathogenicity.


The malate dehydrogenase isoenzymes of Saccharomyces cerevisiae. Purification, characterisation and studies on their regulation. 1. One mitochondrial and one cytoplasmic malate dehydrogenase isoenzyme could be purified from acetate grown cells of the yeast Saccharomyces cerevisiae. 2. The purification procedure uses chromatography on dextran blue columns as an essential step for enrichment, and reverse ammonium sulfate chromatography on celite for isoenzyme separation. 3. The homogeneity of the preparations was established by gel electrophoreses in the presence of sodium dodecylsulfate and by a sedimentation run in the analytical ultracentrifuge. 4. Both enzymes are dimers with a molecular weight of 75 000 for the cytoplasmic and of 68 000 for the mitochondrial enzyme. 5. Amino acid analysis and peptide mapping showed that both enzymes are closely related, but genetically different (true isoenzymes). 6. The cytoplasmic enzyme shows electrophoretic splitting. This is most likely due to post-translational deamination in vivo. 7. Antibodies to both isoenzymes could be obtained in rabbits. The antisera to cytoplasmic malate dehydrogenase were specific for this enzyme. Antisera to mitochondrial malate dehydrogenase react with both isoenzymes. Neither type of antisera precipitated an inactive protein after the glucose-dependent inactivation of cytoplasmic malate dehydrogenase in vivo.


Lipid intermediates in the synthesis of the inner core of yeast mannan. The synthesis by yeast microsomes of compounds that are probably dolichol-pyrophosphate derivatives containing N,N'-diacetylchitobiose and several mannose residues is described. However, the presence of monosaccharide residues other than N-acetylglucosamine and mannose has not been ruled out. The amount of the lipid derivatives synthesized was enhanced by the addition to the incubation mixture of an organic-solvent-soluble extract from rat liver known to contain dolicholpyrophosphate oligosaccharides. Incubation of these derivatives with yeast microsomes led to the transfer of about fifteen percent of their saccharide moieties to endogenous proteins. The oligosaccharides released from the dolichol derivatives by mild acid hydrolysis could serve as primers for the synthesis of a polysaccharide having the characteristics of mannan outer-chain. It is suggested that the dolichol-pyrophosphate derivatives described in this paper are intermediates in the synthesis of mannan inner core.


Simultaneous purification and some properties of aspartate: tRNA ligase and seven other amino-acid:tRNA ligases from Escherichia coli. A procedure is described for the purification of the aspartate:tRNA ligase from Escherichia coli to a stage where it was homogeneous by polyacrylamide gel electrophoresis. From the same batch of E. coli the lysine, phenylalanine and serine ligases were obtained in an apparently homogeneous form while the alanine, glutamine, leucine and valine enzymes had a purity varying from 20% to 80%. Aspartate: tRNA ligase, which has not been obtained in a highly purified form before, has been characterized in terms of its molecular parameters.


Initiation of lambda DNA replication in vitro promoted by isolated P-gene product. The product of the P gene of bacteriophage lambda was isolated from heat-induced lambda-lysogenic Escherichia coli cells. It was found to bind to DNA, to be devoid of nuclease activity acting on double-stranded lambda DNA and of nicking/closing activity. Initiation of lambda DNA replication promoted by the P-gene product in a complementation assay in vitro was sensitive to rifampicin. Sedimentation analysis of the products and their hybridization to separated lambda DNA strands indicate that lambda DNA was formed in a reaction similar to ring-to-ring replication in vivo. The reaction was symmetric from the beginning, i.e. both lambda DNA strands were copied without delay.


Piracetam in the treatment of post-concussional syndrome. A double-blind study. The effect of piracetam, a cyclical derivative of GABA, was compared with that of a placebo in a double-blind study of 60 patients with post-concussional syndrome of 2-12 months' duration. The daily dose of piracetam was 4,800 mg. After 8 weeks of treatment piracetam significantly reduced the occurrence and severity of the following symptoms: vertigo, headache, tiredness, decresed alertness, increased sweating and neurasthenic symptoms. No significant effect was observed on the following symptoms: tremor, orthostatic symptoms, and memory disorders. Side effect were reported by 64% of the patients under piracetam and by 32% under placebo. In the author's opinion, piracetam seems to be a promising new drug for the treatment of post-concussional syndrome.


Intraoperative blood flow in transplanted human kidneys and its relation to subsequent function. The value of intraoperative blood flow measurement during clinical kidney grafting to predict subsequent function was studied in the course of 19 consecutive transplantations. The results indicate that when blood pressure and heart rate of the recipients are within normal limits there exists a significant relationship between renal blood supply and recovery of the kidney function. The operative blood flow into the graft correlated, moreover, with the state of presensitization of the recipients, as determined by antibody-against-panel assays. The possible causes of these relationships are discussed. It is emphasized that actual viability of allotransplanted kidneys depends on both pre-and intraoperative pathophysiological processes.


New developments in the surgical treatment of renovascular arterial hypertension. Surgery of the renal artery and its branches has not developed at the same rate as the progress made in arterial hypertension renovascular studies. Therefore, the percentage of cure is still low, the mortality rate high and the complications frequent. Based on the experiences in renal allo- and autotransplants, on the progress achieved in different fields, such as extracorporeal kidney surgery, on a new way of approach to the spleen's hilus, on the development of microsurgery and on a better knowledge of the biopathology of vascular grafts, new orientations for this type of surgery are supported. No matter which technique is followed, renal hypothermia by arterial perfusion, elimination of the diseased arterial segment, placement of the kidney in the continuity of another arterial system (auto-or splenorenal transplants), substitution of the transperitoneal approach by the retroperitoneal one, and, in complicated cases, the practice of ex situ arterial reconstruction surgery, is considered fundamental. Statistics, following these guidelines, are presented, which indicates that there were no deaths and that the percentage of success is higher than with classic revascularization surgery.


Fibrinolytic degradation products in urine of anticoagulated transplant patients. A study of the level of urinary fibrinolytic split products in 14 renal transplant patients who received oral anticoagulants postoperatively revealed a tendency toward increased levels of fibrinolytic split products that were temporally related to graft rejection. The amount of fibrinolytic split products in the urine was positively related to the severity of rejection as well as to the final status of the allograft. Thus, the urinary level of fibrinolytic split products, as determined by the latex-coated particle test, can be used as an indicator of the course of warfarin-anticoagulated patients after renal allografting.


Tubular defects in patients forming calcium-containing stones. The study of renal function, primarily the tubular function, in 55 patients with idiopathic, recurrent renal stones showed a large number of abnormalities. 49% of the patients, especially the women were affected. The most common defect was a reduced acidification capacity of varying degrees of severity in 64% of the woman and in 20% of the men. Impairment of the acidification capacity of the distal tubule was found in 18% and of the proximal tubule in 11% of the patients. Inability to dilute the urine after water loading was found in 17% and tubular proteinuria also in 17% of the patients. Most patients with an impaired tubular function had a severe stone disease. The defects in acidification and dilution capacity ought to be of pathogenetic importance for stone formation and should be considered in the selection of preventive therapy.


A review of age changes in perceptual information processing ability with regard to driving. Age related changes in the sensory modalities of hearing and vision, along with changes in the information processing abilities of selective attention, perceptual style, and perceptual-motor reaction time, were reviewed in the context of driving behavior. Literature reported, indicated age related changes in these abilities have relevance for the understanding of the driving behavior of the older adult.


Mesenteric hemopoietic colonies. II. Occurrence in mice after transplantation of syngeneic normal bone marrow cells. Syngeneic normal bone marrow cells (BMC) were i.p. transplanted into six different strains of lethally irradiated mice (other than BALB/c). The occurrence of mesenteric hemopoietic colonies (MHC) was determined microscopically. The size and numbers of MHC were augmented when the transplantation period was lengthened. On the tenth day spleen colonies were found in all mice.


Simultaneous assay by six methods of the effect on haemopoietic precursor cells of adriamycin, methyl CCNU, 60Co gamma-rays, vinblastine, and cytosine arabinoside. The spleen colony assay, three agar colony assays and two repopulating assays have been employed to measure the effect of single doses of adriamycin, methyl CCNU, 60CO gamma-rays, vinblastine and repeated injection of cytosine arabinoside on early haemopoietic precursors. The results show that there is no consistent correlation between the effect of these agents on the six assays, and the differences in proliferation rate of the cells contributing to the various assays could not be demonstrated by the phase specific agents vinblastine and cytosine arabinoside. The relatively high survival of granulocyte repopulating ability after adriamycin and methyl CCNU indicates that the capacity for differentiation and maturation can be influenced by cytotoxic agents so that colony assays while estimating the number of progenitor cells surviving do not necessarily provide a reliable guide to the capacity of the cells for restoring haemopoiesis. Comparison of colony formation by normal bone marrow cells with the three agar colony assays showed that the agar diffusion chamber method gave three times as many colonies and a much smaller variation between experiments compared to the in vitro agar assay with endotoxin mouse serum as colony stimulating factor. The thin layer feeder cell method gave an intermediate number of colonies.


A direct assay of granulocytic repopulating ability. A direct method of assay of the 'granulocytic repopulating ability' (GRA) of bone marrow in normal or treated mice, which utilizes determinations of total bone marrow cellularity and peroxidase positive cells, is described. Ten days after transplantation of normal femoral marrow, the total number of marrow granulocytes in the recipients is proportional to the marrow dose in the range 0.01 to 0.03 of a femur. Above 0.01 femur the proportion of granulocytes is constant at about 60% so there is a high correlation between total cellularity and total granulocytes. Assay of erythroid repopulating ability can be performed in the same animals. Using the GRA assay, the results for the dose response curve for cyclophosphamide, and the relationship of GRA to erythroid repopulating ability (ERA) and spleen colony forming cells, are similar to those obtained using the endotoxin technique. The simplicity of the GRA technique enhances the feasibility of multiple simultaneous hematopoietic precursor cell assays.


Proliferative response of clonal acute myelogenous leukemia cells in localized grafts of normal bone marrow stroma to in vivo stimulation of myelopoiesis. Bone marrow from inbred Wistar/Furth (W/Fu) rats, normal and 90% replaced by acute myelogenous leukemia (AML), was transplanted, respectively, to the left and right kidneys of adult W/Fu rats. After a transient period of hypocellularity, AML grafts repopulated to pre-transplant cellularity by day 9, whereas normal bone marrow (NBM) grafts required 25 days for repopulation to pre-transplant cellularity. Grafted leukemia cells remained localized to the kidney for 17 days. In NBM grafts phlebotomy accelerated erythroid proliferation, and intrathoracic inoculation of live Escherichia coli accelerated myeloid proliferation; in AML grafts only E. coli injection increased bone marrow proliferation. There was no morphologically detectable differentiation of W/Fu AML cells. These studies provide evidence of a myelopoietic response of AML blast cells in vivo and present a transplantation technique for comparison of localized grafts of leukemic and normal bone marrow in the same animal.