Bats as laboratory animals.

Bats as laboratory animals.


Lactate dehydrogenase isozymes in the early mouse embryo.


Role of the state agency, as viewed by the clinical pathologist.


Role of the state in clinical laboratories as viewed by the Social Security Administration.


Role of the Public Health Service in the improvement of clinical laboratories.


Role of the state in clinical laboratories.


The clinical laboratory improvement program in New York City. I. Methods of evaluation and results of performance tests.


Laboratory diagnosis of Mycoplasma pneumonia in a public health laboratory.


Effect of glucose on carbon dioxide assimilation and substrate oxidation by Ferrobacillus ferrooxidans. Ferrobacillus ferrooxidans, grown on either elemental sulfur or ferrous sulfate, was able to use either substrate as an energy source for the assimilation of CO(2). In both cases, 0.01 mumole of carbon was incorporated per mumole of oxygen utilized. Glucose inhibited substrate oxidation and CO(2) fixation. Sulfur and iron oxidation were inhibited 5 to 15% and 40 to 50%, respectively, in the presence of 10% glucose. Under the same conditions, CO(2) assimilation was inhibited 50% with elemental sulfur as the energy source, and was almost totally inhibited when ferrous iron was used.


Nature of the skin-reactive principle in culture filtrates prepared from Paracoccidioides brasiliensis. Mycelial and yeast-phase culture filtrates prepared from three strains of Paracoccidioides brasiliensis exhibited equal reactivity in sensitized guinea pigs. Ethyl alcohol-precipitated fractions obtained from the culture filtrates also showed no difference in reactivity between mycelial and yeast phase when tested in sensitized guinea pigs. Chemical analyses of the ethyl alcohol-precipitated fractions revealed the presence of seven aliphatic amino acids in both the mycelial- and yeast-phase products. Glucose, galactose, arabinose, and glucosamine were also detected, but the relative proportions of these sugars were different for the mycelial phase as compared with the yeast phase. Both the mycelial- and yeast-phase ethyl alcohol precipitated fractions contained 2 to 4% nitrogen, but no protein or nucleic acid could be detected. Removal of nitrogen from the ethyl alcohol-precipitated fractions by chloroform extraction resulted in an almost complete loss of skin reactivity, whereas the material recovered from the chloroform, which contained most of the nitrogen, still exhibited almost as much reactivity as was present prior to extraction. A considerable portion of the reducing substances was removed along with the nitrogen by the chloroform extraction, suggesting a strong chemical link between the carbohydrate and the peptide portions of the active moiety. Since no protein was present in the fractions, it was presumed that the active moiety is a glycopeptide.


Release of proteinase from mycelium of Mucor hiemalis. When Mucor hiemalis NRRL 3103 was grown in soybean medium, only a small fraction of the proteinase produced by the organism appeared in the culture filtrate, whereas the bulk of the enzyme was bound to the mycelial surface. Optimal pH of the proteinase ranged from 3.0 to 3.5. Inclusion of sodium chloride or other ionizable salts in the growth medium, however, resulted in the liberation from the mycelium of the loosely bound enzyme as it was formed. Maximal release of proteinase was achieved at a sodium chloride concentration of 0.5 m. The loosely bound proteinase was eluted also from intact resting mycelium by ionizable salts but not by water or by nonionizable substances. The amount of enzyme eluted from the mycelium depended upon the concentration of sodium chloride up to 0.3 m. Since liberation took place rapidly even at 0 C, a loose ionic linkage must exist rather than a biochemical binding of the enzyme to the mycelium. The recovery of proteolytic activity from repeated salt extractions was greater than that originally detected in the intact mycelium, possibly owing to unmasking of more active enzymes or functional groups. Further proteinase activity was released when salt-extracted mycelium was ruptured. Part of the proteinase thus observed was firmly attached to the cell fraction, and part of it appeared in the supernatant fluid. These conditions implied the presence of intracellular or firmly attached proteinase which could be partially released.


Isolation and characterization of glycolipids from some photosynthetic bacteria. Glycosyl glycerides have been found in substantial amounts in Chloropseudomonas ethylicum but could not be detected in two strains of Rhodopseudomonas palustris. Rhodospirillum molischianum possibly contains small amounts of monoglycosyl diglyceride. The glycolipids of C. ethylicum have been separated into two components. One of these, glycolipid I, is a monogalactosyl diglyceride. Glycolipid II, upon acid hydrolysis, yields galactose, rhamnose, and a third, unidentified sugar. The glycolipids or total lipids of the photosynthetic bacteria examined contained saturated and monounsaturated, but none of the more highly unsaturated, fatty acids.


Aliphatic hydrocarbons and fatty acids of some marine and freshwater microorganisms. Gas chromatography and combined gas chromatography-mass spectrometry have been used to study the fatty acids and hydrocarbons of a bacterium from the Pacific Ocean, Vibrio marinus, a freshwater blue-green alga, Anacystis nidulans, and algal mat communities from the Gulf of Mexico. Both types of microorganisms (bacteria and algae) showed relatively simple hydrocarbon and fatty acid patterns, the hydrocarbons predominating in the region of C-17 and the fatty acids in the range of C-14 to C-18. The patterns of V. marinus were more comparable to those of the algal populations than to patterns reported for other bacteria. An incomplete correlation between fatty acids and hydrocarbons in both types of organisms was observed, making it difficult to accept the concept that the biosynthesis of hydrocarbons follows a simple fatty acid decarboxylation process.


Degradation of hydrocarbons by members of the genus Candida. II. Oxidation of n-alkanes and l-alkenes by Candida lipolytica. Candida lipolytica ATCC 8661 was grown in a mineral-salts hydrocarbon medium. n-Alkanes and 1-alkenes with 14 through 18 carbon atoms were used as substrates. Ether extracts of culture fluids and cells obtained from cultures grown on the various substrates were analyzed by thin-layer and gas-liquid chromatography. Analyses of fluids from cultures grown on n-alkanes indicated a predominance of fatty acids and alcohols of the same chain length as the substrate. In addition, numerous other fatty acids and alcohols were present. Analyses of saponifiable and nonsaponifiable material obtained from the cells revealed essentially the same products. The presence of primary and secondary alcohols, as well as fatty acids, of the same chain length as the n-alkane substrate suggested that attack on both the methyl and alpha-methylene group was occurring. The significance of these two mechanisms in the degradation of n-alkanes by this organism was not evident from the data presented. Analyses of fluids from cultures grown on 1-alkenes indicated the presence of 1,2-diols, as well as omega-unsaturated fatty acids, of the same chain length as the substrate. Alcohols present were all unsaturated. Saponifiable and nonsaponifiable material obtained from cells contained essentially the same products. The presence of 1,2-diols and omega-unsaturated fatty acids of the same chain length as the substrate from cultures grown on 1-alkenes indicated that both the terminal methyl group and the terminal double bond were being attacked.


Biochemical and genetic analysis of isoleucine and valine biosynthesis in Staphylococcus aureus. After a prototrophic strain of Staphylococcus aureus had been exposed to diethyl sulfate, 28 isoleucine- and isoleucine-valine-dependent mutants (ilv mutants) were isolated. On the basis of auxanography, their ability to accumulate intermediates of isoleucine and valine biosynthesis, and intergeneric syntrophism with ilv mutants of Salmonella typhimurium, all mutants were placed into four groups, each of which corresponded to a presumed enzymatic deficiency, as follows: group A, deficient in l-threonine deaminase; group B, deficient in the condensing enzyme; group C, deficient in reductoisomerase; group D, deficient in alpha-beta-dihydroxy acid dehydrase. No mutants blocked in the terminal (transaminase) reactions were isolated. Transduction analyses (best-fit, ratio, and complementation tests) with the use of phage 83 established that the linear arrangement of the structural genes is identical with the order of participation of their enzymes in isoleucine and valine biosynthesis, and that these genes comprise a single linkage group which can exist on a single donor fragment during transduction.


Phospholipids and glycolipids of sterol-requiring Mycoplasma. The phospholipids of Mycoplasma hominis type 2 strain 07 are composed almost entirely of phosphatidyl glycerol. Traces of other glycerophospholipids may exist. No glycolipids are found. The phospholipids of Mycoplasma sp. avian strain J are composed of diphosphatidyl glycerol, which predominates in older cultures, a monoacyl glycerophosphoryl glycerophosphate, which may serve as a precursor of diphosphatidyl glycerol, and phosphatidyl glycerophosphate. This organism also contains cholesteryl glucoside and an unidentified glycolipid which appears to be similar to a monoglucosyl diglyceride. No turnover or radioisotope labeling of the phospholipids occurs during metabolism. This lack of turnover during growth is indicative of a structural role for these glycerophospholipids. A concomitant decrease of monoacyl glycerophosphoryl glycerophosphate and increase of diphosphatidyl glycerol occurs during growth.


Altered protein formation as a result of suppression in Neurospora crassa. The mutant td201 of Neurospora crassa is mutated in the trp-3 locus and forms an altered tryptophan synthetase. A suppressor mutation, su2-6, in this mutant, unlinked to the trp-3 locus, results in the production of wild-type tryptophan synthetase activity, which accounts for the alleviation of the tryptophan or indole requirement. This enzyme activity is associated with a protein physically dissimilar to the wild-type enzyme. A second altered protein, a serologically cross-reacting material is also formed in the suppressed mutant, in addition to the altered enzyme normally formed by the td201 mutant. Normal growth, equivalent to that of wild type, is not restored in the suppressed mutant even with tryptophan supplementation. The relationship of the data to possible mechanisms of suppression is discussed.


Ultrastructure of the cell wall of Bacillus polymyxa. The macromolecular arrangement on the surface of Bacillus polymyxa was revealed by metal shadowing of whole cells and wall fragments; it consisted of a rectangular array of 70-A globules with a repeating interval of 100 A. The substructure was studied in plan with phosphotungstic acid (pH 6) or uranyl acetate as negative stains of fragments and was studied also in profile with sections of embedded material. Staining of sections of cells fixed with glutaraldehyde showed that layering (approx. 80-A dense, 40-A light, and 120-A dense layers, outermost layer first) could be demonstrated in the cell wall with lead or uranyl acetate, used together or separately. The outer "dense" layer corresponded to the regularly arrayed structure (RS); it was removed by guanidine hydrochloride, sodium lauryl sulfate, cold formamide, and by trypsin. The RS layer (isolated by a hydrogen bond breaking reagent, guanidine hydrochloride) was disrupted by agents such as sodium lauryl sulfate or damaged by 3 m sodium chloride. Qualitative chemical tests, ultraviolet absorption, and removal by trypsin indicated that the structured layer consisted mainly of protein, but exact characterization was not attempted. The globular units making up the layer consisted of a small number of subunits, imperfectly resolved by negative staining. The underlying polysaccharide appeared to be covalently bound to the deepest (probably mucopeptide) layer since it required "hot" formamide for its removal. A survey of species was not made.


Inhibition and agglutination of arthrobacters by psuedomonads. The bacterial flora of water in Narragansett Bay, R.I., was observed semimonthly from 1962 to 1964. Dominant isolates were keyed to genus, and the isolates for each genus were expressed as percentage of total isolates. There was a consistent inverse relationship between arthrobacters and the dominant pseudomonads. Pseudomonad growth on agar plates markedly inhibited arthrobacter cross-streaks. Agar from inhibition zones as well as supernatant fluids from pseudomonad broth cultures inhibited arthrobacter motility and caused the cells to agglutinate. Gummy pseudomonad residues from vacuum-evaporated broth cultures readily passed a G-25 Sephadex column. This material agglutinated arthrobacter cells, but failed to cause arthrobacter inhibition in filter-pad assays. In contrast, sterile medium inside a dialysis sac, inoculated externally with a pseudomonad, was inhibitory to arthrobacters in pad assay but failed to agglutinate arthrobacter cells. Pseudomonad isolates from soil showed similar inhibiting and agglutinating activities for both soil and seawater arthrobacter isolates. The inhibitory and agglutinating activities of pseudomonad isolates appeared to diminish on prolonged laboratory cultivation.


Fatty acid synthetase of Saccharomyces cerevisiae. A light particle fraction of Saccharomyces cerevisiae, obtained from the crude ribosomal material, and containing the fatty acid synthetase, consisted primarily of 27S and 47S components. This fraction has a protein-ribonucleic acid ratio of about 13. Electron micrographs showed particles ranging in diameter between 100 and 300 A in this material. By use of density gradient analysis, the fatty acid synthetase was found in the 47S component. This component contained particles which were predominantly 300 A in diameter and which were considerably flatter than ribosomes, and it consisted almost entirely of protein.


Electron microscopy of the lysis of Staphylococcus aureus cell walls by Aeromonas lytic factor. Isolated and purified cell walls of Staphylococcus aureus were treated with a purified fraction of the culture supernatant fluid of a species of Aeromonas. The course of lysis of the cell walls was followed over a period of time by examination of samples under an electron microscope. The undifferentiated cell wall was rapidly digested, but the equatorial rings were more resistant. The undifferentiated cell wall became a very thin sheet before completely dissolving, leaving a series of equatorial rings of various widths. As digestion proceeded, solubilization of the entire cell wall occurred. Analogous findings were obtained with purified S. aureus mucopeptide. It is concluded that the Aeromonas lytic principle is an enzyme, and that susceptible bonds are more concentrated in the undifferentiated cell wall mucopeptide.


Purification and chemical composition of reticulate bodies of the meningopneumonitis organisms. Reticulate bodies of the meningopneumonitis (MP) microorganism were purified from L cells 18 hr after infection by the combination of differential centrifugation in 30% sucrose solution and potassium tartrate density gradient centrifugation. It was ascertained by electron microscopy that purified preparations of reticulate bodies obtained were almost entirely free of host-cell components and of infectious elementary bodies of MP microorganisms. Purified reticulate bodies were easily disrupted by mechanical agitation, and it was observed in shadowed preparation that ribosome-like particles 15 mmu in diameter were scattered from broken reticulate bodies. In shadowed preparations, reticulate bodies were found to range in size from 1.0 to 1.6 mu in diameter, but in cross-section the range was 0.5 to 1.0 mu. In these preparations, the purified reticulate bodies were irregular in shape, round or oval, and were composed of rather homogenous, amorphous, or reticulate material with moderate density. Some particles exhibited a less-dense internal structure, in which a coarse fibrous reticulum was seen. Chemical fractionation of (32)P-labeled purified reticulate bodies showed that they contained three times more ribonucleic acid (RNA) than deoxyribonucleic acid, with the RNA being composed primarily of 21S, 16S, and 4S RNA. No infectivity of purified reticulate bodies could be demonstrated.


Isolation of ribosome particles from meningopneumonitis organisms. In ribonucleic acid (RNA) extracted by phenol and sodium dodecyl sulfate from purified reticulate bodies of meningopneumonitis (MP) organisms, 21S, 16S, and 4S RNA were found by sucrose density gradient sedimentation analysis. When purified reticulate bodies were homogenized by sonic treatment or by treatment with sodium deoxycholate and were fractionated by differential centrifugation, more than 50% of the RNA was recovered in the fraction which was sedimented by centrifugation at 105,000 x g for 2 hr, but not at 13,000 x g for 20 min. From homogenates prepared in this manner, 50S and 30S particles containing RNA were isolated by sucrose density gradient centrifugation. These 50S and 30S particles were also found in lysates of cytoplasmic fractions of infected cells which were labeled by (32)P during 17 to 17.5 hr or 15 to 18 hr after infection. The synthesis of 50S and 30S particles was not inhibited by actinomycin D. When infected cells were homogenized in the presence of 0.01 or 0.02 m MgCl(2), 70S particles were isolated instead of 50S and 30S particles. When dialyzed against low concentrations of MgCl(2), the 70S particles dissociated to 50S and 30S particles. The base ratio of the 70S particles is very similar to that of 16S plus 21S RNA. The characteristics of the 70S, 50S, and 30S particles suggest that these are ribosome particles, similar to bacterial ribosomes.


Isolation of thymidine diphosphate D-glucose, thymidine diphosphate D-galactose, and thymidine diphosphate 4-acetamido-4, 6-dideoxy-D-galactose from Pasteurella pseudotuberculosis.


Effect of succinate on isocitrate lyase synthesis in Mima polymorpha.


Physical behavior of streptolysin S.

Physical behavior of streptolysin S.


Influence of pH on the inhibitory activity of formic and acetic acids for Shigella.


A unique defense mechanism against anthrax demonstrated in dwarf swine.


Partial biochemical characterization of the activated esterase required in the complement-dependent chemotaxis of rabbit polymorphonuclear leukocytes. It was shown in the preceding paper that incubation of the rabbit polymorphonuclearleukocyteswith phosphonate esters leads to an irreversible inhibition of the ability of the leukocyte to respond to the chemotactic factor. This "cell-dependent inhibition" was attributed to the inactivation by the phosphonates of an esterase existing in or on the leukocyte in an already activated state. As shown in this paper, incubating the leukocyte with phosphonate in the presence of certain esters prevents this cell-dependent inhibition. The protection is specific; the ester must be an acetate. Ethyl formate, ethyl propionate, ethyl butyrate, glucose 6-phosphate, fructose 1,6-diphosphate, ATP, tosyl arginine methyl ester, or acetyl tyrosine ethyl ester do not protect. The protection is independent of the phosphonate used to inhibit, and the degree of protection depends on the relative concentrations of acetate and phosphonate. Those acetates which protect are also the esters which inhibit chemotaxis when added to the leukocyte in the upper part of the chemotaxis chamber. It is concluded that the activated esterase is an enzyme capable of specifically splitting, or binding acetates, or doing both. Presumably the esterase is some type of acetylesterase or acetylase. The known aliesterase present in the leukocyte is not the activated esterase. Inhibition of the activated esterase by phosphonates has no effect on endogenous or exogenous glycolysis.


The effect of cross-reacting antigens on the tolerant state. Rabbits were rendered tolerant to human albumin (HA) and were then injected with azo and oxazolonated derivatives of human albumin. These injections were continued to a time at which all animals would have lost tolerance if they had not been injected. Injection of cross-reacting antigens prolonged the duration of tolerance, as judged by the mode of elimination of lightly iodinated human albumin (HA.(131)I). Different derivatives of HA differed in their capacity to prolong tolerance. Those neonatally injected rabbits which were immunized with cross-reacting antigens and lost tolerance, responded much more promptly to HA.(131)I than animals which were not immunized. Animals immunized with cross-reacting antigen which went on to eliminate HA.(131)I triphasically, usually had responded earlier by making antibodies. These antibodies contained a fraction which was reactive with HA, and which was usually equally well adapted to determinants on HA and on the cross-reacting antigen.


Immunochemical studies on the group and type antigens of group F streptococci and the identification of a grouplike carbohydrate in a type II strain with an undesignated group antigen. Two antigens, the group-specific carbohydrate and the Type II carbohydrate, have been isolated by cellulose column chromatography from a formamide extract of a Group F streptococcus. Chemical and immunologic analyses indicate that both antigens are free of other cellular components. Both antigens are components of the cell wall although the Type II antigen is probably more superficial than the group antigen. The Type II antigen is composed of rhamnose, glucose, galactose, and galactosamine. The Group F antigen is composed of rhamnose, glucose, galactosamine and a small percentage of glucosamine. A grouplike carbohydrate and the Type II carbohydrate have been isolated from a streptotoccal strain which lacks a serologically detectable streptococcal group antigen. This grouplike carbohydrate, which does not cross-react immunologically with Group F serum, is composed of rhamnose, galactose, and glucosamine. No chemical or immunological differences were observed between the Type II antigen isolated from the Group F strain and the Type II antigen isolated from the nongroupable strain.


The effect of passively administered antibody on antibody synthesis. Suppression of the primary response of rabbits to intravenously administered KLH can be achieved with very small amounts of hyperimmune anti-KLH administered a day later since the rabbit apparently rapidly eliminates most of the KLH by nonimmunologic means. The amount of passive anti-KLH needed to achieve immunosuppression was directly proportional to the dose of injected antigen. Antibody passively administered as much as 6-8 days after antigen still can be strongly immunosuppressive, which suggests that the antibody must be reacting with immunogen in or on responding cells or perhaps in the process of transfer between cells. There was no evidence that the presence of passively administered hyperimmune anti-KLH prior to the injection of antigen had any immunosuppressive action beyond the direct neutralization of the injected antigen. When KLH was injected in Freund adjuvant, anti-KLH incorporated with the KLH in the adjuvant was much more efficient in causing immunosuppression than anti-KLH given intravenously. The primary responses to 2 mg KLH given intravenously and 2 microg given in adjuvant reached approximately equal peaks and were suppressible by comparable amounts of intravenously administered anti-KLH. Two observations suggest that passive antibody neutralizes the immunogenic stimulus at the level of individual antigenic determinants and not merely by aggregating or precipitating entire antigenic molecules. First, anti-abalone hemocyanin (AH) which cross-reacts approximately 50% with KLH was only partially immunosuppressive even in extremely large amounts, i.e., amounts which could react with and precipitate much more KLH than could the smaller but more suppressive doses of anti-KLH. Second, when KLH and anti-KLH were given together in adjuvant, effective immunosuppression was achieved only with amounts of anti-KLH sufficient to saturate or cover virtually all available antigenic determinants. The immunosuppressive quality of passive antibody increases with time after immunization and with repeated immunization of the donor. In view of their relatively weak immunosuppressive properties, antibodies formed in the first weeks of a primary response may not contribute significantly to the turning off of the antibody response. In any event, results obtained by passive transfer of hyperimmune antibody to animals early in a primary response cannot be applied to the natural events in a primary response.


Primary immune response in grafted cells. Dissociation between the proliferation of activity and the proliferation of cells. The primary immune response elicited by host antigens in a grafted population of immunologically competent cells has been compared in conditions where the same dose of parental cells were grafted simultaneously to F(1) hybrid embryos of 13 or 17 days of age. The enlarged chimeric spleens harvested 4 days later were analyzed for donor cell proliferation by using the sex chromosomes as karyological markers, and for proliferation of immunological activity by means of transfer to secondary hosts of the same genotype. Whereas the total number of dividing donor cells were on the average 16 times higher in 17-day than in 13-day hosts, the recovery of immunological reactivity showed a 6- to 7-fold difference in the opposite direction. The experiments cast doubt on the proposition that cellular proliferation is necessary for development of a primary immune response. They suggest that there exists an alternative way in which a primary immune response may unfold from involving a few to involving a much larger number of cells.


Hypertrophic spinal pachymeningitis.

Hypertrophic spinal pachymeningitis.


On the nature of transplantation immunity in the adenovirus tumor system. The existence of a virus-induced, virus-specific transplantation, antigen in adenovirus 12-induced CBA mouse tumors was demonstrated. The antigen is virus-specific, but not related to structural virion or T antigens. It is a weak antigen, and required immunization with whole, infectious adenovirus 12 to produce considerable immunity. Comparable immunity could not be achieved with homologous cellular or subcellular materials, but some indication of enhancement was produced with low tumor dose. Immunization required at least 2 wk and was mediated by immune lymphoid cells. Serum of immunized animals showed no demonstrable cytotoxicity or enhancement. Animals immunized with virus and Freund's adjuvant showed diminished transplantation immunity, although these animals were actively immunized against adenovirus type 12 structural virion antigens.


The immune response to foreign red blood cells and the participation of short-lived lymphocytes. The sequence of morphological changes in the rat spleen following SRBC injection associated with hemolysin production has been correlated with estimates of proliferative activity by splenic lymphatic tissue. Formation of new, reactive germinal centers containing macrophages which engulf nuclear debris is a prominent feature of the response. This is prevented by pretreatment of the animal with cortisol. Indirect evidence is provided that short-lived lymphocytes produced in germinal centers may be a necessary component in the induction of other cells to proliferate and differentiate into hemolysin-producing cells. The reasons are discussed for considering short-lived lymphocytes, such as those produced in the thymus, bone marrow, and germinal centers, as differing from long-lived lymphocytes capable of antibody synthesis.


Coarctation of the aorta and cerebral aneurysm. Report of two cases.


Experimental evaluation of four synthetic adhesives for possible treatment of aneurysms.


Neoplastiform xanthomatous granulomas of choroid plexus in a child affected by Hand-Schuller-Christian disease. Case report.


Total removal of large acoustic neuromas. A modification of the McKenzie operation with special emphasis on saving the facial nerve.


Development of extracranial metastases from a malignant astrocytoma in the absence of previous craniotomy. Case report.


Experience in autologous and allogenetic thyroid grafts.


Continuous recording of the intracranial internal carotid artery size in the dog.


Erythropoietic responses to acute and chronic ureteral occlusions in dogs.


Serial massive hepatic resections in cirrhotic rats. Changes in in vitro growth of liver cells.


Problems in animal procurement.

Problems in animal procurement.


Endocytosis in thyroid follicle cells. II. A microinjection study of the origin of colloid droplets.


Electron microscopic alterations of submaxillary gland produced by isoproterenol.


Electron microscope studies on some modifications of the plasmalemma in oranges.


Electron microscopic studies on the distal border of the canine olfactory epithelium.


Fine structure of the development of the cephalopod lens.


Ultrastructure of hemocyanin components from the gastropod Buccinum undatum.


Hemoglobin absorption in the proximal tubules of the kidney in the rabbit.


Microtubular elements and cell membranes isolated from nucleated erythrocytes.


Structural and chemical asymmetry of the calcium-transporting membranes of the sarcotubular system as revealed by electron microscopy.


A prospective study of South Australian blood donors. 2. First annual review.


An organization for intensive care.

An organization for intensive care.


Haematological indices in a random sample survey of the highlands region of the territory of Papua and New Guinea.


Vitamin B 12 absorption studies--the effect of carbachol and gastrin on the 57-Co-labelled vitamin B 12 plasma level absorption tests in patients with pernicious anaemia.


The philosophy of a medical scientist.

The philosophy of a medical scientist.


Response to treatment in adult coeliac disease and the paneth cell.


Smallpox eradication.

Smallpox eradication.


Genetics in medical practice. 2.

Genetics in medical practice. 2.


An experiment in paramedical education.

An experiment in paramedical education.


Diverticulum of the female urethra.

Diverticulum of the female urethra.


Activation of thromboplastin in a case of abruptio placentae with hypofibrinogenemia.


Clotpromoting components in circulating blood in a case of abruptio placentae with afibrinogenemia.


The coagulation mechanism in oral contraception.


Tuberculous hepatitis with jaundice in pregnancy.


Paracervical block.

Paracervical block.


Double-contrast hysterosalpingography for the visualization of intrauterine contraceptive devices.


Spinal changes in patients with congenital aplasia of the vagina.


Juvenile ankylosing spondylitis.

Juvenile ankylosing spondylitis.


The visceral lesions in rheumatoid arthritis.


Juvenile rheumatoid arthritis. Introduction to the Symposium.


Controlled clinical trial of flufenamic acid in rheumatoid arthritis.


Crossed extensor reflex action upon cyclic muscle stretch.


Growth hormone secretion provoked by insulin-induced hypoglycaemia in children of short stature.


Evaluation of a training leg for above-knee amputees.


Growth hormone-induced nitrogen retention in children of short stature.


Chronic diarrhoeas in infancy and childhood. II. Enzyme content of duodenal juice.


The "happy puppet" syndrome.

The "happy puppet" syndrome.


Pulmonary stenosis, café-au-lait spots, and dull intelligence.


Side-effects to antibiotics in cystic fibrosis: dental changes in relation to antibiotic administration.


Pulmonary veno-occlusive disease.

Pulmonary veno-occlusive disease.


Addison's disease and diabetes mellitus.


Difficulty in swallowing, with aspiration pneumonia in infancy.


Serum lipoproteins in schoolboys in relation to glucose-6-phosphate dehydrogenase deficiency and thalassaemia trait.


Higher oxides of nitrogen as an impurity in nitrous oxide.


The chemistry of the higher oxides of nitrogen as related to the manufacture, storage and administration of nitrous oxide.


Higher oxides of nitrogen in anaesthetic gas circuits.


Two cases of poisoning by contamination of nitrous oxide with higher oxides of nitrogen during anaesthesia.


The chemistry of the in vivo reaction between haemoglobin and various oxides of nitrogen.


Morbid anatomical changes in the lungs of dogs after inhalation of higher oxides of nitrogen during anaesthesia.


Detection and determination of higher oxides of nitrogen.


Current methods of commerical production of nitrous oxide. A contribution from The British Oxygen Co. Ltd.


Entries relating to nitrous oxide in the British pharmacopoeia since 1932 (which was the first year the gas received mention).


Manufacture of nitrous oxide. Contribution from Kingston Chemicals Limited.


Extended pyelolithotomy for renal calculi.


Treatment of urinary tract infections in children with nalidixic acid.


Testosterone-induced activation of fibrinolysis in carcinoma of the prostate.


Nephrectomy for hypertension due to unilateral renal disease.


Accurate surgical localisation of renal calculi.


Renal arteriovenous fistula. A report of four cases and review of the literature.


Arteriovenous fistula associated with renal cell carcinoma of the kidney. Report of three cases with cardiovascular findings.


Perforation as a complication of carcinoma of the bladder.


Proliferative changes in bladder mucosa and carcinoma in situ.


A method of training in perurethral resection.


Urethrocystography in the male.

Urethrocystography in the male.


The "hourglass" calculus: vesico-prostatic calculus following prostatectomy.


Prepubertal gynaecomastia in association with an interstitial-cell tumour of the testis.


Malignant phaeochromocytoma of the organ of Zuckerkandl with functioning metastases.


Input impedance of the systemic circulation.


Effect of respiration on venous return and stroke volume in cardiac tamponade. Mechanism of pulsus parodoxus.


Enhancement of coronary vasodilator action of adenosine triphosphate by dipyridamole.


Improved assay methods for renin "concentration" and "activity" in human plasma. Methods using selective denaturation of renin substrate.


Interactions of quinidine and potassium on atrioventricular transmission.


Genetic transmission of congenital membranous ventricular septal defects in selectively inbred substrains of rats.


Effects of stimulation frequency on myocardial extensibility.


Fats and fatty acids in excess.

Fats and fatty acids in excess.


Calculated and analyzed fatty acids in vegetarian diets.


Diet and heart disease in the cardiovascular health center.


Changes in amino acid permeation during sporulation. Changes in amino acid uptake in Bacillus licheniformis and in the amino acid pools of three Bacillus species were investigated, by use of cells from different stages of the life cycle. B. licheniformis contains catalytic uptake systems for all of the 10 amino acids studied. The apparent maximal velocities of uptake decreased during sporulation but did not fall below the range observed for other microorganisms. In sporulating cells, the apparent affinity constants of the uptake systems for individual amino acids remained about the same as in growing cells, i.e., from 2 x 10(-7)m to 7 x 10(-6)m, whereas, in some cases, the apparent maximal velocities decreased significantly. Because the velocity of uptake showed an atypical dependence on substrate concentration, it was postulated that these cells contain two or more uptake systems for each amino acid. Only one of these systems appeared to be operative at a substrate concentration below 10(-6)m. Working at these low substrate concentrations, catalytic activities producing a net efflux of amino acids were demonstrable in vegetative cells in the presence of chloramphenicol, but these exit systems were lost during sporulation. A pool formed by the addition of radioactive algal hydrolysate will exchange with the external medium in vegetative cells but not in sporulating cells. Glutamic acid and alanine comprise at least 60% of the amino acid pool of B. licheniformis A-5, B. subtilis 23, and B. cereus T during all stages of growth and sporulation. The concentrations of the other amino acids in the pool varied extensively, but reflected, in general, the amino acid turnover known to occur during sporulation.


Regulation of glutamine synthetase. II. Patterns of feedback inhibition in microorganisms. The feedback inhibition of glutamine synthetase was investigated by use of partially purified enzyme preparations from Salmonella typhimurium, Micrococcus sodonensis, Pseudomonas fluorescens, Bacillus cereus, Bacillus licheniformis, Clostridium pasteurianum, Rhodospirillum rubrum, Neurospora crassa, Candida utilis, and Chlorella pyrenoidosa. Inhibition analyses indicated that the enzyme of each organism can be effectively regulated with mixtures of end products from the diverse pathways of glutamine metabolism. When tested individually, tryptophan, histidine, alanine, glycine, glutamine, 5'-adenylate (AMP), cytidine-5'-triphosphate, carbamyl phosphate, and glucosamine-6-phosphate gave limited inhibition. In most cases, the inhibitors were independent in their action, and cumulative degrees of inhibition were obtained with mixtures of these end products. In contrast, with the glutamine synthetases of the two Bacillus species, the simultaneous presence of AMP and histidine (or AMP and glutamine) gave inhibition greater than the sum of the amounts of inhibition caused by either inhibitor alone. Also, alanine and carbamyl phosphate acted synergistically to inhibit the enzyme from N. crassa. The remarkable similarity in the overall patterns of end-product inhibition observed with the enzymes from different sources indicates that these diverse organisms have evolved comparable mechanisms for the regulation of glutamine metabolism. Nevertheless, the enzymes from different sources do differ significantly in their physical and catalytic properties, as was demonstrated by dissimilarities in their purification behaviors, specificity for nucleotide substrate, ability to catalyze the glutamyl transfer reaction, and ability to utilize Mn(++) and Mg(++) as activators for the biosynthetic reaction.


Enzymatic oxidation of tetramethyl-p-phenylenediamine and p-phenylenediamine by the electron transport particulate fraction of Azotobacter vinelandii. The ability of the electron transport particulate fraction of Azotobacter vinelandii strain O to oxidize tetramethyl-p-phenylenediamine (TMPD) and p-phenylenediamine (PPD) was examined in detail. The highest specific activity for TMPD and PPD oxidation concentrated in the A. vinelandii O R(3) fraction. The A. vinelandii O R(3) fraction was used to develop a standard manometric assay which gave optimal oxidation rates for both of these dyes. The conditions of the assay and all essential related enzymatic kinetic parameters are presented. Other para derivatives of phenylenediamines also were oxidized readily, whereas ortho and meta derivatives were not. Hydroquinone, p-hydroxybenzoic acid, p-cresol, tyrosine, pyrogallol, pyrocatechol, and diphenylamine were not able to serve as electron donors for the A. vinelandii O R(3) system. The probable involvement of a particle-bound cytochrome oxidase is indicated by the marked sensitivity of both TMPD and PPD oxidation to cyanide, axide, phenylhydrazine, hydroxylamine, and, to a lesser degree, carbon monoxide.


Physiology of rickettsiae. VI. Host-independent synthesis of polyribonucleotides by Coxiella burnetii. Preparations of purified and disrupted suspensions of Coxiella burnetii are able to incorporate ribonucleotides into polymers in the presence of adenosine, guanosine, cytidine, and uridine triphosphates. Nucleotide incorporation requires the presence of all four ribonucleoside triphosphates. The reaction is enhanced by the addition of phosphoenolpyruvate and pyruvic kinase, and exogenous deoxyribonucleic acid, and is inhibited by deoxyribonuclease and actinomycin D. Incorporation is maximal between pH 7.0 and 8.0, and at 37 C. The synthesized polymer is relatively insensitive to deoxyribonuclease and is sensitive to ribonuclease and dilute alkaline hydrolysis. The data indicate the presence of an autonomous deoxyribonucleic acid-dependent ribonucleic acid polymerase in the rickettsial agent.


The cell wall of Rickettsia mooseri. I. Morphology and chemical composition. Cell walls prepared by mechanically disrupting intact Rickettsia mooseri (R. typhi) were examined in an electron microscope and analyzed chemically. Electron micrographs of metal-shadowed and negatively stained rickettsial cell walls revealed no significant differences, except for smaller size, from bacterial cell walls prepared in a similar manner. The chemical composition was complex, and resembled that of gram-negative bacterial cell walls more closely than that of gram-positive bacterial cell walls. R. mooseri cell walls contained the sugars, glucose, galactose, and glucuronic acid, the amino sugars, glucosamine, and muramic acid, and at least 15 amino acids. Diaminopimelic acid, a compound hitherto found only in bacteria and blue-green algae, was demonstrated in rickettsiae for the first time. Teichoic acids were not detected. The compounds identified accounted for about 70% of the dry weight of the cell walls.


Indentification and localization of the fatty acids in Haemophilus parainfluenzae. Haemophilus parainfluenzae was capable of synthesizing 22 fatty acids. These fatty acids were equivalent to 4% of the bacterial dry weight. These fatty acids were localized in the membrane-wall complex, which contained the respiratory pigments, the quinone, and the phospholipids. The fatty acids which could be extracted with organic solvents comprised 86% of the total fatty acids of the cell. These fatty acids were distributed as 98% in the phospholipids and 1.9% in the neutral lipids, of which 0.5% were free fatty acids. Palmitic, palmitoleic, oleic, and vaccenic acids comprised 72% of the total fatty acids and were found almost exclusively in the phospholipids. The phospholipids also contained the cyclopropane fatty acids. The neutral lipids contained significant proportions of the odd-numbered branched and straight-chain fatty acids. The principal free fatty acids were n-dodecanoic and pentadecenoic acids. The nonextractable wall complex contained 14% of the total fatty acids. These wall fatty acids were rendered soluble only after saponification. The wall fraction contained all of the beta-hydroxymyristic acid and most of the myristoleic and pentadecenoic acids. The significance of the distribution of fatty acids between nonesterified, neutral lipid, phospholipid, and nonextractible wall remains to be determined.


Growth of Mycobacterium lepraemurium in cultures of mouse peritoneal macrophages. Successful growth of Mycobacterium lepraemurium was observed in cultures of mouse peritoneal macrophages. The optimal host cell maintenance medium was composed of 40% horse serum, 50% of the chemically defined medium NCTC 109, and 10% of a 1:5 dilution of beef embryo extract, supplemented with both liver extract and ferric nitrate. Multiplication of the bacilli was observed in 1 week and maximal growth in 6 to 7 weeks. All macrophages were filled with tens to hundreds of the organisms in cultures showing maximal growth. Glycerol caused an increase in the normal length of M. lepraemurium, without a corresponding increase in the number of the bacilli. Elongation of M. lepraemurium was observed 3 or 4 days after infection. Rapid and uniform growth of M. lepraemurium was achieved in serially transferred cultures (subcultures). The cumulative increase of the number of intracellular bacilli was 1.4 x 10(20)-fold in 14 transfers over a period of 68 weeks in one series, and 10(17)-fold in 12 transfers over a period of 56 weeks in another series. The generation time of M. lepraemurium was 7 days, a growth rate which approximates the fastest growth of the organisms in vivo. Organisms harvested from cultures at various stages of growth produced murine leprosy in mice, but showed no growth in bacteriological media. The present model offers an opportunity for studies on the host-parasite relationship without the complication of extracellular growth of the parasites.


Large bodies of Mycoplasma and L-form organisms. The large bodies of various Mycoplasma and L-form organisms were studied by ultraviolet fluorescence microscopy of preparations stained with various fluorochromes. Primuline and Thioflavine S specifically stained the outer portion or rim of the large bodies, and the fluorescence characteristics of the stained bodies differed from those for other microorganisms and for spheroplasts and protoplasts. Small granular structures similar in size and morphology to minimal reproductive units were observed within some of the large bodies by phase microscopy and by fluorescence microscopy with acridine orange or Coriphosphine O. Micromanipulation probing of the large bodies revealed their elastic nature; many of the large bodies could be subdivided into two or more smaller circular bodies, each retaining the fluorescence staining properties of the parent body. Under these conditions, however, a few of the large bodies were ruptured, leaving the stainable outer boundary area as a stable residual structure. The large bodies were somewhat resistant to various rigorous treatments normally employed to eliminate viability of Mycoplasma and L-form cultures. Structures similr to large bodies were observed in various natural tissues, and structures resembling large bodies in size, morphology, fluorescence staining characteristics, and reaction to micromanipulation probing were reconstructed from an acetone extract of egg yolk. Overall, the large bodies of Mycoplasma and L-form organisms appeared to be structures resulting from accumulations of metabolic by-products and medium components within or on which minimal reproductive units had become entrapped, although it could not be ruled out that they might be defined structures specifically formed during culture as protective lipoidal sacs for the minimal reproductive units.


Clostridium spores with ribbon-like appendages. Spores of Clostridium sp. N1 are characterized by numerous broad ribbon-like appendages attached to one end. The appendages are two to three times the length of the spore and, at their maximal dimension, may be two-thirds the width of the spore. They are attached to the spore body by a common trunk which is continuous with the outer spore coat. Each appendage is a multilayered structure and is enclosed in an amorphous material. Details of spore and appendage formation are described, and appendage ultrastructural features are presented. The function of the appendages is not known.


Mesosomes in m-tyrosine-inhibited Bacillus thuringiensis.


Immobilization of certain cultured treponemes in sera from syphilitic humans.


Role of carotenoids and cholesterol in the growth of Mycoplasma laidlawii.


Ionic remediability of a mutational transport defect in Chlamydomonas.


Lysis of Staphylococcus aureus by culture supernatant fluids of a species of Aeromonas.


Comparison of the virulence for chickens of lyophilized and of cultured Mycobacterium avium.


Measurement of bacteriolytic enzymes.

Measurement of bacteriolytic enzymes.


Development of resistance to novobiocin, tetracycline, and a novobiocin-tetracycline combination in Staphylococcus aureus populations. The antibiotic sensitivity of the individual organisms of a bacterial population was determined to study the comparative rates of development of resistance of Staphylococcus aureus to novobiocin, tetracycline, and to a combination of these antibiotics. Serial subculture of S. aureus with the combination of novobiocin-tetracycline (N-T 2.5:1; the ratio in serum of patients dosed with Panalba) showed a significant retardation of resistance outgrowth compared with subculture in the presence of the antibiotics individually. Increase in organisms resistant to novobiocin seen after one N-T subculture was related to the "concentration gap" between novobiocin and tetracycline. Two additional subcultures with N-T caused little or no increase in organisms resistant to novobiocin, tetracycline, or to the combination. The data suggest that the retardation of further development of resistance was the result of tetracycline inhibition of novobiocin-resistant strains and vice versa.


Evidence of lactate dehydrogenase in Anaplasma marginale. Extracts from preparations of partially purified Anaplasma marginale revealed low levels of lactate dehydrogenase (LDH). Enzyme inhibition by immune sera further indicated that A. marginale possesses a protein moiety the same as that of the normal red blood cell (RBC), although data suggested an alteration of LDH(1) from that observed in normal RBC. Bimodal isozyme distribution was detected after electrophoresis of the extracts. One isozyme approached the cathode and the other the anode, and both appeared to be nicotinamide adenine dinucleotide-dependent. Heterogeneity of parasite and host cell isozymes was established on the basis of zone electrophoresis on cellulose acetate strips.


Effects of valine on protein synthesis and turnover in Pseudomonas saccharophila under "nongratuitous" inducing conditions. Under "nongratuitous" inducing conditions, in Pseudomonas saccharophila, d-and l-valine and l-isoleucine inhibit net protein synthesis. At a concentration of 0.5 mumole or greater of valine per mg of bacterial protein, net protein synthesis declined approximately 70%. The inhibitory effect of valine is proportional to the exogenous valine concentration. Studies of (14)C amino acid incorporation and (14)C amino acid release from prelabeled cells indicate that valine stimulates protein turnover.


Yield coefficients of Thiobacillus neapolitanus in continuous culture. Thiobacillus neapolitanus, when grown in continuous culture with thiosulfate limiting growth, possessed an apparent maximal molar growth yield of 8.0 g (dry weight) per mole of thiosulfate. The substrate requirement for energy of maintenance was the highest yet reported, amounting to 21.8 mmoles of thiosulfate per g per hr. The molar growth yield, corrected for this maintenance energy requirement, was 13.9 g (dry weight) per mole of thiosulfate. It was concluded that substrate-level phosphorylation during sulfite oxidation accounted for about 45% of the adenosine triphosphate (ATP) requirement for CO(2) assimilation and maintenance during growth on limiting thiosulfate, that three sites of energy conservation exist in the electron-transport chain terminating in oxygen, and that 7.8 moles of ATP are required to fix and assimilate 1 mole of CO(2) into cell material.


Purification and properties of Streptococcus lactis beta-galactosidase. beta-Galactosidase of Streptococcus lactis 7962 was partially purified, and its properties were studied. Enzyme from only this strain of numerous lactic streptococci tested was stable in cell exudates prepared by various means. Cell-free extracts of the 7962 strain were prepared by sonic treatment of washed cells previously grown in presence of lactose to fully induce enzyme synthesis. Protamine sulfate precipitation of the nucleic acids and ammonium sulfate precipitation of protein were used for partial purification of the enzyme. The resulting enzyme, when resuspended in cold (5 C) phosphate buffer, was extremely labile. However, ammonium sulfate in high concentrations (0.85 m) stabilized and stimulated beta-galactosidase activity. Sephadex G-200 gel filtration was used to achieve further purification and to monitor homogeneity of the enzyme. Separation of the beta-galactosidase in buffer at 5 C yielded an enzyme elution pattern showing two peaks of activity. However, addition of the enzyme solution in 0.85 m ammonium sulfate to the column equilibrated with the same salt concentration yielded only one peak of enzyme activity. The data suggested that the native enzyme was dissociating into active subunits which were stabilized in the presence of the ammonium sulfate.


Use of analogues and the substrate-sensitivity of mutants in analysis of purine uptake and breakdown in Aspergillus nidulans. Aspergillus mutants resistant to various purine analogues (purine, 8-azaguanine, 2-thioxanthine, and 2-thiouric acid) are defective in at least one step of purine uptake or breakdown. The properties of these mutants show that there are two uptake systems for purines, one which mediates the uptake of hypoxanthine, guanine, and adenine, and the other, xanthine and uric acid. Allantoinase-less strains are sensitive to the toxic effects of allantoin accumulation. They are severely inhibited when grown in the presence of naturally occurring purines. Mutant strains derived from these, resistant to naturally occurring purines, may be isolated. These are either wild-type revertants, or carry a second metabolic block in the uptake or breakdown of purines. The properties of these double mutants confirm the interpretation of the nature of the analogue-resistant mutants.


Metabolic injury to bacteria. II. Metabolic injury induced by distilled water or Cu++ in the plating diluent. When distilled water from a tin-lined still served as the plating diluent, cells of Aerobacter aerogenes developed symptoms of metabolic injury as evidenced by increased counts on supplemented, as compared with minimal, plating medium. Cysteine was as effective as yeast extract as a supplement to the minimal medium in increasing the viable count. Mg(++) and, to a lesser extent, phosphate buffer at the concentrations tested protected unfrozen cells, but not cells which had been frozen and stored, against the loss of capacity to grow on minimal medium. When the plating diluent consisted of distilled water redistilled in an all-glass still, the symptoms of metabolic injury did not appear. Spectrographic analysis revealed the presence of 10(-7)m Cu(++) in the distilled water, and Cu(++) added to redistilled water serving as the plating diluent reproduced the metabolic injury effects induced by distilled water. It was concluded that freezing and storage damaged the cell membrane, rendering it more penetrable by toxic elements which were thereby enabled to act at sites in the cell where Mg(++) and other solutes in the plating diluent could not serve as effective antagonists. Increased recovery of cells on supplemented medium could be ascribed to the capacity of the supplements to remove toxic elements which had become bound to the cells during suspension in the plating diluent.


Mode of filamentous growth of Leucothrix mucor in pure culture and in nature, as studied by tritiated thymidine autoradiography. Mode of growth of Leucothrix mucor filaments was measured by autoradiography with tritiated thymidine. Studies were performed on L. mucor in pure cultures in free suspension, as an epiphyte of pure cultures of the red alga Antithamnion sarniense, and as an epiphyte of red algae in the sea. Statistical analyses of the distribution of growing cells was done by use of the nonparametric One-Sample Runs Test and a Cluster analysis adapted from quadrat analyses of plant ecologists. No evidence of preferential growth at base or tip of L. mucor filaments was obtained in any of these studies. However, in nature, but not in the laboratory, there were regions of L. mucor filaments which were nongrowing or dormant. Such nongrowing regions could incorporate tritiated glucose.


Comparative glucose catabolism of Xanthomonas species. Glucose catabolism in eight Xanthomonas species has been comparatively examined by means of the radiorespirometric method. The basic mechanisms for the respective xanthomonads closely resembled each other. The Entner-Doudoroff pathway, in conjunction with the tricarboxylic acid cycle pathway, was the predominant mechanism for glucose catabolism. A small portion (8 to 16%) of substrate glucose was routed into the pentose phosphate pathway. The hexose cycle pathway did not appear to play any significant role in glucose catabolism of these xanthomonads. The results are also consistent with the well-recognized close phylogenic relationship between xanthomonads and pseudomonads.


Influence of urea on the growth of T-strain mycoplasmas. T-strain mycoplasmas require urea for propagation, but urea metabolism also occurs in nonpropagating viable cultures. Ammonia results from this metabolism and alkalinizes the medium. Ammonium ions and an alkaline pH both inhibit the multiplication of T strains and reduce the viability of T strains in broth. These toxic effects of urea metabolism currently limit the growth of T strains in broth. Stock T-strain cultures are optimally maintained in continuous culture if the routine medium at pH 6.0 is supplemented with 0.05% urea and 0.002% phenol red, but an incubation temperature of 30 C is preferable to 37 C for subculture at 24-hr intervals.


Immunosuppressive activity of the 3a-1 fraction of papain. The maximal nonlethal dose of the 3a-1 fraction of papain, determined by use of 9-week-old white albino rabbits, was 10 mg per injection, administered intravenously. The immunosuppressive activity of the 3a-1 fraction of papain was studied by its inhibition of sheep red blood cell hemolysin, bacterial agglutinin, and horse serum precipitin. Immune suppression was observed when papain injections preceded antigen injections by 12 to 18 hr. The enzyme preparation was analyzed by use of cellulose acetate and starch-gel electrophoresis and by the micro-Kjeldahl method. Starch-gel electrophoresis of serum samples revealed qualitative alterations in the alpha(-1) region 7 hr after papain administration. Changes were also observed in the urinary aminopolysaccharide content.


Physiology of rickettsiae. VII. Amino acid incorporation by Coxiella burnetii and by infected hosts. Protein synthesis, in terms of (14)C-labeled amino acid incorporation into a hot trichloroacetic acid fraction, was studied in cell-free preparations of Coxiella burnetii, and in uninfected and Q fever-infected guinea pig and chick embryo hosts. Purified and disrupted suspensions of C. burnetii incorporated (14)C-labeled l-leucine, l-phenylalanine and algal hydrolysate. Livers of infected guinea pigs and chick embryos had a greater incorporation rate at the height of infection than comparable preparations from uninfected animals. As chick embryonic development continued during infection, the rate of incorporation progressively decreased below that of uninfected embryos.


Occurrence of urease in T strains of Mycoplasma. A previously unknown metabolite necessary for growth of T strains of Mycoplasma in artificial culture media has been identified as urea. The source of this metabolite was the mammalian plasma or serum enrichment of the culture medium. Normal horse serum was the most satisfactory native protein enrichment for cultivation of T strains of mycoplasma, and it is believed that its superior performance in agar and fluid culture media is associated with its relatively high urea content (approximately 40 mg/100 ml). T-strain urease activity was maximal at pH 6.0 +/- 0.5. This is also the optimal pH for growth of T strains. Substrate concentrations greater than 1.0% urea were inhibitory to growth and urease activity of T-strain organisms, and optimal urea concentrations in fluid media appeared to lie within the range of 0.008 to 0.01 m. This range of urea concentration permitted maximal growth of T-strain organisms without rapid loss of viability due to excessive ammonia accumulation and rise in pH to lethal levels. T strains of Mycoplasma were cultivated in a serum-free fluid medium containing urea as the only added metabolite and nitrogen source. T strains are the only known human mycoplasmas which exhibit urease activity, and this biochemical marker can be employed as an aid in the detection and identification of T strains of Mycoplasma (urease color test) and in distinguishing T strains from other members of the human Mycoplasma group.


Stability of ribosomes and ribosomal ribonucleic acid from Bacillus stearothermophilus. After heating at 65 C, ribosomes isolated from Bacillus stearothermophilus were strikingly more heat-stable than comparable preparations from Escherichia coli when tested for ability to support polyuridylic acid-directed phenylalanine incorporation at 37 C. The stability of ribosomes was also determined by measurements of hyperchromicity at 259 mmu while heating them from 25 to 90 C. In standard buffer containing 0.01 m Mg(++), the T(m) (temperature at the midpoint of total hyperchromicity) of E. coli and B. stearothermophilus ribosomes was 71 and 81 C, respectively. In a magnesium-free buffer, the T(m) of E. coli and B. stearothermophilus ribosomes was 44 and 64 C, respectively. Putrescine (0.01 m) was more effective in stabilizing ribosomes from B. stearothermophilus than those from E. coli. Spermidine (0.001 m), on the other hand, was more effective in stabilizing ribosomes from E. coli than those from B. stearothermophilus. Melting curves of total ribosomal ribonucleic acid (rRNA) from E. coli and B. stearothermophilus revealed T(m) values of 50 and 60 C, respectively. Putrescine stabilized thermophile rRNA, but had no effect on E. coli rRNA. Sucrose density gradients demonstrated that thermophile 23S ribonucleic acid was degraded during storage at -20 C; the 23S component from E. coli was stable under these conditions. The results are discussed in terms of the mechanism of ribosome heat stability and the role of the ribosome in governing the temperature limits for bacterial growth.


Changes in adenine nucleotides of intact Chromatium D produced by illumination. The total adenine nucleotide content of suspensions of Chromatium D averaged 14 nmoles/mg of dry weight. Of this, one-third to one-half was adenosine triphosphate (ATP), even in suspensions incubated in darkness. Illumination with high intensities caused a rise in ATP and a drop mainly in adenosine diphosphate, the new steady state being reached in 5 to 15 sec at room temperature. The dark steady state was re-established 15 to 30 sec after returning the suspensions to darkness. The rates of these changes were little affected by the presence of electron donors or CO(2), though their magnitude was reduced when substrates were added to starved suspensions. At limiting light intensities, complex kinetics characterized the transition from both dark to light and light to dark, and, at lower light intensities, more ATP was produced in suspensions supplemented with electron donors than in starved cells. The results show that photophosphorylation accompanying cyclic electron flow occurred in intact cells, and suggest that noncyclic phosphorylation can also occur.


Experimental salmonellosis. 8. Postinfective immunity and its significance for conferring cellular immunity. In the process of live-vaccine immunization of Salmonella enteritidis infection in mice, the relation between the number of bacteria in the organs of mice and their protecting effect was studied. Treatment with antibiotics was used to control the number of immunizing bacteria in the tissues. Mice, which were infected with 10(-5) mg (1,000 mouse MLD) of virulent S. enteritidis and treated with kanamycin simultaneously, acquired high antilethal resistance against infection with the same organisms. However, the administration of large amounts of kanamycin, which caused a rapid decrease in bacterial numbers in the organs of infected mice, was incapable of conferring immunity. This indicated the necessity of persistence of live bacteria in the host for the production of immunity. A large number of microorganisms were maintained for 53 weeks in a diffusion chamber inserted into the mouse abdominal cavity. The mice implanted with diffusion chambers containing large numbers of virulent S. enteritidis did not acquire antilethal resistance against infection with the same organisms, although agglutinins against S. enteritidis were observed in these mice. Agglutinin was also found in the fluid contained in diffusion chambers inserted into mice immunized with a killed vaccine of S. enteritidis. This indicated that antibody penetrated the membrane filter of diffusion chambers from outside to inside and vice versa. From these results, it is suggested that contact of live microorganisms with the host cell is necessary for conferring postinfective immunity in salmonellosis.


Effect of diethylaminoethyl dextran on the growth of Mycoplasma in agar. The growth of certain strains of Mycoplasma is inhibited by substances present in commercial agar preparations. The addition of diethylaminoethyl (DEAE) dextran (10 mg per 100 ml) to agar media appears to enhance the growth of some strains. Of eight strains initially tested, the presence of DEAE dextran grossly enhanced the growth of three strains. One strain appeared not to be affected, and a clearly enhancing effect was not evident with four strains. Quantitative studies revealed that growth enhancement varied from 10 colony-forming units (CFU) for M. hominis type II (strain Campo) to 10(3.3) CFU for M. pulmonis (strain 880). The growth-enhancing effect is probably due to the ability of DEAE dextran to bind the sulfated polysaccharide moieties in agar and not to the DEAE dextran, per se.


Effect of L-methionine and S-adenosylmethionine on growth of an adenine mutant of Saccharomyces cerevisiae. A pink, adenine-requiring yeast utilized adenine, hypoxanthine, or S-adenosylmethionine (SAM), in quantities up to 3 mumoles per 100 ml of medium, as equivalent sources of purine for cell growth, but not methylthioadenosine or S-adenosylhomocysteine. Utilization of SAM for growth was inhibited by the presence of l-methionine in quantities greater than 0.6 mumole per 100 ml of medium. However, 6 mumoles of l-methionine had no effect on growth when adenine or hypoxanthine was the source of purine. These sources also reversed the inhibitory effects of 6 mumoles of the amino acid on the utilization of SAM. The presence of 400 mumoles of the amino acid resulted in some inhibition of growth when the organisms were grown with adenine, hypoxanthine, or adenine plus SAM but had no effect on the total uptake of adenine-8-(14)C. Studies on the uptake of radioactivity from a mixture of SAM-adenine-8-(14)C and (3)H-labeled SAM-methyl indicated that these components were taken into the cells at different rates which were altered by the presence of l-methionine. The fixation of (35)S from (35)S-labeled adenosylmethionine into the cells was inhibited by the presence of the amino acid. The cells synthesized and accumulated SAM in the presence of 400 mumoles of l-methionine plus adenine even when exogenous SAM was supplied. Approximately 47% of radioactivity fixed from exogenous SAM-adenine-8-(14)C and 12% from (3)H-labeled SAM-methyl were found in reisolated SAM.


D-arabinose countertransport in Bakers' yeast. The initial rate of the glucose-induced countertransport of d-arabinose was measured at several concentrations of extracellular glucose. These data permit the calculation of the intracellular concentration of free glucose, and, if the rate of glucose metabolism is known, the maximal rate of glucose transport can be estimated. Since the maximal transport rate remained essentially constant when the extracellular glucose concentration was increased from 2 to 100 mm, the results are consistent with the hypothesis that, during glucose metabolism, glucose is transported across the yeast cell membrane by a symmetrical carrier system which functions independently of metabolism.


Growth of pathogenic Leptospira in chemically defined media. A protein-free chemically defined medium for cultivation of pathogenic Leptospira was developed. The medium permitted continued serial subculturing of 9 serogroups (52 strains) of the 12 serogroups (61 strains) tested. Growth was initiated from small inocula, and the growth rate and maximal cell yields were similar to those on serum-containing media. The nutritional requirements of serogroups L. canicola, L. pomona, and L. grippotyphosa were studied in a basal medium composed of inorganic salts, a fatty acid, vitamin B(12), and thiamine. All strains tested utilized ammonium chloride as the sole nitrogen source. A fatty acid, vitamin B(12), and ferrous ions were essential. Growth was stimulated by thiamine, potassium, and calcium ions.


Comparative inductive responses of two beta-galactosidases of Neurospora. The activities of the two beta-galactosidases of Neurospora increased in response to the presence of lactose or galactose. d-Xylose preferentially induced higher levels of activity in one of these enzymes. The initial period of the inductive response was examined in cultures transferred from noninducing to inducing media. Variations in the ratio of activities of these enzymes during this induction period indicate that their induction was not coordinate. The induction of the beta-galactosidases of Neurospora was less sensitive to low concentrations of inducers than was the induction of the beta-galactosidase of bacteria; the time lag of induction was greater in the eucaryotic organism, and the maximal induction of active enzyme was less than that observed in bacteria.


Evidence for multiple forms of isocitrate lyase in Neurospora crassa. The effect of carbon source on isocitrate lyase formation was studied in a wildtype strain of Neurospora crassa and in a uridine-deficient mutant. A constitutive level of the enzyme was produced in a casein hydrolysate medium. The enzyme was repressed by glucose, although the two strains varied with respect to the degree of glucose repression. Acetate strongly stimulated isocitrate lyase formation. The enzyme formed in the presence of acetate differed in several respects from that formed in glucose-grown cells. Differences were found in pH-activity curves, K(m) values, and in sensitivity to phosphoenolpyruvate inhibition. Diethylaminoethyl cellulose chromatography allowed separation of two enzymatically active components which showed different rates of heat inactivation. These data indicate the presence of multiple forms of isocitrate lyase in Neurospora.


Alteration of the pathogenicity of Pasteurella pneumotropica for the murine lung caused by changes in pulmonary antibacterial activity. Pasteurella pneumotropica is a potential pulmonary pathogen in mice. In healthy animals, this organism was killed rapidly by the normal function of the intrapulmonary phagocytic defense mechanisms. Impairment of this bactericidal activity by the acute renal failure of nephrectomy resulted in multiplication of the Pasteurella in the lung, both when the animals were nephrectomized first and then infected, and when the animals were infected first and nephrectomized several hours after the infection. The study demonstrates that the pathogenicity of the Pasteurella organisms is governed by the functional state of these pulmonary antibacterial mechanisms.


Carbohydrate metabolism in leukocytes. VII. Metabolism of glucose, acetate, and propionate by human plasma cells. Plasma cells obtained from the peripheral blood of a patient with multiple myeloma was incubated in serum and Krebs-Ringer bicarbonate buffer with (14)C-labeled glucose, acetate, and propionate. Glucose utilization by these cells amounted to 0.5 mumole per hr per 10(8) cells and was mainly via the Embden-Meyerhof pathway, and only 6% or less traversed the hexose monophosphate shunt. The presence of Krebs cycle activity was demonstrated by direct isolation of several labeled intermediates after incubation with either (14)C-acetate or (14)C-propionate. The distribution of (14)C in lactate, succinate, fumarate, malate, aspartate, and glutamate indicate a complete Krebs cycle. Acetate was metabolized via the Krebs cycle to the extent of 0.15 mumoles per hr per 10(8) cells, and the rate of propionate utilization was 0.17 mumoles per hr per 10(8) cells.


Control of lysine biosynthesis in yeast by a feedback mechanism. Homocitric acid (beta-hydroxy-beta-carboxyadipic acid; HC) is accumulated by a lysine-requiring yeast mutant when grown in a chemically defined medium, supplemented with limited amounts of lysine. A study of the formation of HC in relation to the depletion of lysine from the growth medium indicates that HC accumulated only when the concentration of lysine was low. The enzymatic formation of HC from alpha-ketoglutarate plus acetyl-coenzyme A in cell-free extracts of the same organism was also inhibited by lysine. The inhibitory effect of lysine on the formation of HC in both whole cells and cell-free extracts is indicative of the functional existence of a feedback control mechanism in the pathway for lysine biosynthesis in yeast.


Occurrence of 3-amino-3,6-dideoxyhexoses in Salmonella and related bacteria. Several species of Salmonella, Citrobacter, and Arizona were examined for the presence of 3-amino sugars, which were isolated from lipopolysaccharide hydrolysates by cation exchange chromatography and identified by paper and cation exchange chromatography in several systems and by specific colorimetric procedures. 3-Amino-3,6-dideoxyglucose was identified in C. freundii 8090, C. freundii 869, S. halle, S. telaviv, S. dakar, S. wandsworth, and S. champaign; 3-amino-3,6-dideoxygalactose was found in S. tranoroa and Arizona 24. Evidence suggests that these 3-amino sugars occur in lipopolysaccharides as the N-acetyl derivatives. The composition of the lipopolysaccharides containing the 3-amino sugars was determined, and the lipopolysaccharides were compared serologically in order to correlate chemotypes with serotypes. The lipopolysaccharides containing 3-amino-3,6-dideoxyglucose reacted with serogroups 28 or 39; those containing 3-amino-3,6-dideoxygalactose were found to react with group 55 antisera. The preparation of a lipopolysaccharide from the phenol phase of the 44% aqueous phenol extraction procedure is also reported.


Culture of serum-induced spheroplasts from Vibrio cholerae. Spheroplasts of Vibrio cholerae types Inaba, Hikojima, and Ogawa were produced in serum. An excess of lysozyme was added to expedite transformation, and CaCl(2) was used as the stabilizing agent. At the optimal time for each strain, when less than 1% classical rod forms were observed microscopically, samples were plated on both conventional and modified L-form media. No growth occurred on conventional media; L-form type colonies appeared on L-form media after 2 to 6 days of incubation. L-form colonies could be subcultured on conventional media and required from four to six passages for complete reversion to classical parent forms. Reaction mixtures of Hikojima and Inaba types were passed through membrane filters (0.45 mu); L-form colonies were grown from both strains after spheroplast transformation. Appropriate controls were negative. It is suggested that the replicative particle may be filtrable.


Methane utilization by a strain of Rhodopseudomonas gelatinosa.


Growth and ultrastructure of Rhodomicrobium vannielii as a function of light intensity. Cells of Rhodomicrobium vannielii were grown in a controlled environment at several different light intensities. Differential rates of bacteriochlorophyll (BChl) synthesis and specific BChl contents were inversely related to the light intensity. On the other hand, the specific rate of growth-before reaching a maximal value-was directly related to the intensity of the light. Thin sections of cells grown at moderately low light showed the typical peripherally located, symmetrically distributed lamellate system, whereas an asymmetrical distribution of a less extensive lamellate system occurred in cells grown at high light intensities. It is proposed that a limited number of individual units of the lamellate system are originally derived from inward folds of the cytoplasmic membrane, and that subsequent lamellae arise by proliferation, including possible forking and definite folding back, of the few original lamellar membranes.


Energetics and motility in Bacillus licheniformis.


Fatty acid composition of spores of the "thermophilic anaerobes".


Glycolytic activity of Streptococcus mitis grown in vitro and in gnotobiotic animals.


Mycotoxin from Penicillium puberulum.

Mycotoxin from Penicillium puberulum.


Isolation of a pregnenolone metabolite, delta-5-pregnene- 3-alpha, 16-alpha, 20-alpha-triol, from the urine of normal men.


Characterization of a thyroid-stimulating factor in a human cancer tissue.


Plasma estriol levels during normal spontaneous labor and labor induced by oxytocin infusion.


Indications and operative procedure for portocaval anastomosis.


Air-induced complications of open heart operations. An experimental study.


Late complications following implantation of vascular prosthetics.


Quantitative influence of blood volume expansion on the osmotic threshold for vasopressin release.


Mean plasma concentration, metabolic clearance and basal plasma production rates of testosterone in normal young men and women using a constant infusion procedure: effect of time of day and plasma concentration on the metabolic clearance rate of testosterone.


Caution in the use of phosphates in the treatment of severe hypercalcemia.


Studies on enzyme concentration and adaptation in human liver and adipose tissue.


Failures and complications observed in 34 patients treated with temporary and permanent electric cardiac stimulation.


A clinical and hemodynamic evaluation of the results of surgical correction of atrial septal defect.


Clinical experience with the implantation of epicardial and intracardial pacemakers.


Cortisol binding capacity of plasma transcortin in diabetic subjects.


Studies by electron microscopy on the effects of magnesium chloride-procaine amide or potassium citrate on the myocardium in induced cardiac arrest.


Dependence of thyroidal clearance rate on plasma iodide level.


The role of "leakage" of tubular fluid in anuria due to mercury poisoning. The role of "leakage" of tubular fluid in anuria produced by mercury poisoning was studied in rats by micropuncture techniques. After an initial brisk diuresis, almost all animals were completely anuric 24 hours after HgCl(2) injection. Lissamine green injected intravenously in the early stage of anuria appeared in the beginning of the proximal tubule, but the color became progressively lighter as the dye traversed the proximal convolutions. The dye was barely visible in the terminal segments of the proximal tubule; it did not appear at all in the distal tubules. These observations suggest that the proximal epithelium had become abnormally permeable to Lissamine green. Tubular fluid to plasma inulin (TF/P(In)) ratios and inulin clearance were measured in individual nephrons at three sites: early proximal tubule, late proximal tubule, and distal tubule. It was found that TF/P(In) ratios were abnormally low in the late proximal and distal tubules. Inulin clearance was normal at the beginning of the proximal tubule but fell by more than 60% by the late proximal convolutions. Thus, the proximal tubule had also become permeable to inulin. We conclude from these observations that anuria in mercury poisoning can occur in the presence of a normal glomerular filtration rate. The absence of urine flow appears to be due to complete absorption of the filtrate through an excessively permeable tubular epithelium. The driving force affecting this fluid absorption is probably the colloid oncotic pressure of the peritubular capillary blood.


Renin release during acute reduction of arterial pressure in normotensive subjects and patients with renovascular hypertension. In normotensive subjects, acute reduction of mean arterial pressure to from 60 to 75 mm Hg by infusion of sodium nitroprusside caused significant increase in renin activity of renal venous plasma and also in the renal-systemic difference of renin activity. At the same time, the products of the renal-systemic difference of renin activity and renal plasma flow increased significantly, whereas renin substrate activity of plasma was unchanged, indicating that there was an increase in renin release during reduction in pressure. Renin activity of renal venous plasma, expressed in logarithms, showed a significant correlation with the degree of reduction in pressure; an increase in renin activity became significant when mean arterial pressure was reduced to below a level of 70 to 75 mm Hg. There was a striking difference in the renal response to reduction in pressure between patients with renovascular hypertension and normotensive subjects. In 10 renovascular hypertensive patients, significant increase in renin release occurred from the involved kidney at mean arterial pressures ranging from 90 to 137 mm Hg; the threshold at which renin release increased was shifted to a range much higher than that in normotensive subjects. Furthermore, the magnitude of renin release from the involved kidney was significantly greater when compared to that in normotensive subjects. In contrast, in the contralateral uninvolved kidney, no significant release of renin was detected during reduction in pressure. The renal mechanism controlling renin secretion appears to be operative at higher systemic arterial pressure levels and with enhanced responsiveness in the involved kidney of renovascular hypertensive patients; the findings are consistent with the hypothesis that the renin-angiotensin system participates in maintaining hypertension in this disease.


A comparison of the metabolism of radioactive 17-isoaldosterone and aldosterone administered intravenously and orally to normal human subjects. After intravenous and oral administration of radioactive aldosterone to normal subjects, 7.3 +/- 0.4 (SE) and 5.4 +/- 0.5 (SE)%, respectively, of the dose was recovered from a 48-hour collection of urine as aldosterone released by mild acid hydrolysis (from aldosterone 18-glucuronide), and 35 +/- 5 (SE) and 39 +/- 4 (SE)%, respectively, was recovered as tetrahydroaldosterone after incubation with beta-glucuronidase.After intravenous and oral administration of 17-isoaldosterone-4-(14)C to a similar group of subjects, 35 +/- 3 (SE) and 53 +/- 4 (SE)%, respectively, of the dose was recovered as 17-isoaldosterone released by acid and less than 5% as total metabolites after incubation with beta-glucuronidase. No detectable radioactivity (< 0.5%) could be recovered as tetrahydroaldosterone or as a compound with the expected chromatographic properties of tetrahydro-17-isoaldosterone. The total radioactivity in the neutral extracts was also relatively small (< 2%) after administration of either labeled aldosterone or 17-isoaldosterone. The radioactivity as aldosterone in the neutral extract was much lower after oral [0.017 +/- 0.003 (SE)%] than after intravenous [0.21 +/- 0.04 (SE)%] administration of labeled aldosterone. The radioactivity as 17-isoaldosterone in the neutral extract was similar after intravenous [0.20 +/- 0.02 (SE)%] and after oral [0.38 +/- 0.18 (SE)%] administration of 17-isoaldosterone. These results indicated that, due to lack of A-ring reduction of the molecule and the consequent slowing of hepatic clearance, 17-isoaldosterone is converted to an acid-labile conjugate (presumably 17-isoaldosterone 18-glucuronide) as the major metabolite. 17-Isoaldosterone was not secreted or converted to aldosterone to any significant extent in the normal subjects investigated.


The steroid content of adrenal adenomas and measurements of aldosterone production in patients with essential hypertension and primary aldosteronism. The content of aldosterone, corticosterone, and cortisol has been assayed in normal adrenal tissue obtained at autopsy and in adrenal adenomas from both normotensive and hypertensive patients obtained at autopsy and from patients with primary aldosteronism obtained at surgery. The content of aldosterone and corticosterone in the adenomas of patients with essential hypertension was similar to that of normal adrenal tissue and much less than that of adenomas from patients with primary aldosteronism. Since these results do not appear to reflect various extraneous factors such as assay technique, degradation of steroids, and patient selection, they suggest that most adrenal adenomas found in patients with essential hypertension are not associated with increased aldosterone production. The presence of such adenomas should not, in itself, be considered evidence for primary aldosteronism. The excretion or secretion of aldosterone in 43 hypertensive patients was found to be similar to that of 39 normotensive patients. These results suggest that primary aldosteronism is rarely the cause of "essential" hypertension.


Thyrocalcitonin, EGTA, and urinary electrolyte excretion. The infusion of thyrocalcitonin (TCT) into thyroparathyroidectomized rats, given either no exogenous parathyroid hormone or a constant infusion of this hormone, leads to a transient phosphaturia and a decreased excretion of urinary magnesium, calcium, and hydroxyproline without a change in glomerular filtration rate. The changes in phosphate excretion may be due to a direct effect of the hormone upon renal tubular function or they may be a consequence of the fall in plasma calcium brought about by the action of TCT upon bone. In support of this latter alternative is the fact that the infusion of sodium ethylenebis-oxyethylenenitrilotetraäcetic acid (EGTA, a specific chelator of calcium) also leads to phosphaturia presumably as a consequence of hypocalcemia. However, EGTA infusion leads to enhanced urinary hydroxyproline excretion and sustained phosphaturia. These latter observations are interpreted to mean that alterations in the local ionic environment of osteolytic cells lead to changes in their activity and constitute a local regulatory system whose activity is modulated by the hormones, thyrocalcitonin and parathyroid hormone.


Serum-red cell interactions at low ionic strength: erythrocyte complement coating and hemolysis of paroxysmal nocturnal hemoglobinuria cells. Complement coating and hemolysis were observed when erythrocytes from patients with paroxysmal nocturnal hemoglobinuria (PNH) were incubated in isotonic sucrose solution in the presence of small amounts of serum. Normal cells were likewise coated with complement components but did not hemolyze. Both normal and PNH erythrocytes reduced the hemolytic complement activity of the serum used in this reaction. Experience with other simple saccharides and related compounds suggests that the low ionic strength of the sucrose solution is the feature that permitted complement coating of red cells and hemolysis of PNH erythrocytes. Isotonic solutions of other sugars or sugar alcohols that do not readily enter human erythrocytes could be substituted for sucrose. The mechanism for these reactions may possibly relate to the agglutination observed with erythrocytes tested in the serum-sucrose system. Even though PNH hemolytic activity could be removed by prior heating of serum or barium sulfate treatment of plasma, the agglutination phenomenon still persisted. The in vitro conditions necessary for optimal sucrose hemolysis of PNH erythrocytes were described and compared with those of the classical acid hemolysis test. The requirement for less serum in the sucrose hemolysis system than needed in the standard acid hemolysis reaction makes certain experiments, especially those using large amounts of autologous PNH serum, much more feasible. Additional advantages of the sucrose hemolysis test are that it can be carried out at room temperature in the presence of oxalate and citrate and that critical pH control is not essential. To date, the sucrose hemolysis test has been a sensitive and specific one for PNH. A modified test used for screening purposes, the "sugar water" test, is very easy to perform.


Basal plasma glucagon levels of man.

Basal plasma glucagon levels of man. The isolated perfused rat liver can serve as a bioassay system for glucagon, capable of detecting 10 mmug of this agent. Seven 15-ml plasma specimens obtained from different healthy volunteers after an overnight fast were assayed in this system; glucagon could not be detected in any of them, indicating concentrations significantly below 0.67 mmug per ml in all subjects. The effects of administering small doses of glucagon to patients were consistent with these results; imposition of increments to plasma glucagon concentration below 1 mmug per ml induced distinct and sustained increases in blood glucose. Observations of the biologic effects of glucagon, together with data on the rate of its inactivation by the liver, suggest that the basal concentration of this hormone in peripheral plasma probably does not exceed 0.1 mmug per ml.


Studies on the coumarin anticoagulant drugs: interaction of human plasma albumin and warfarin sodium. In studies by continuous flow electrophoresis the coumarin anticoagulant drug warfarin sodium was found to be bound solely to the albumin fraction of the plasma proteins. The interaction was studied in detail by equilibrium dialysis of solutions of crystalline human plasma albumin and warfarin sodium. Analysis of the data showed that albumin possesses a single strong binding site for warfarin with an association constant of 154,000 at 3 degrees C and secondary classes of several sites with a much lower affinity. The free energy of binding for the first anion determined at 3 degrees and 37 degrees C was -6.54 and -7.01 kcal per mole, respectively. The standard enthalpy change for the interaction was -3.48 kcal per mole, and the entropy change was +11.2 U. The negative enthalpy change was surprisingly large and the positive entropy change small for an anion-albumin interaction, suggesting significant nonionic binding. The inability to saturate the albumin binding sites, even when high concentrations of warfarin were used, is consistent with a reversible configurational alteration of the albumin molecule during the binding process. The thermodynamic data indicate that the albumin binding sites for warfarin sodium are formed during the process of binding, rather than being performed as in antigen-antibody reactions. The strength of the binding process suggests that many of the pharmacodynamic characteristics of warfarin sodium in man are determined by its strong interaction with plasma albumin. Such correlations of the physicochemical interactions and biologic effects of the coumarin anticoagulant drugs should lead to a better understanding of their mechanisms of action.


The dissociation by colchicine of phagocytosis from increased oxygen consumption in human leukocytes. The effect of colchicine on the uptake of oxygen by human leukocytes during phagocytosis of live streptococci or of killed staphylococci was compared with the effect of colchicine on phagocytosis per se, measured in a sensitive bacterial system. The increase in oxygen consumption that normally accompanies phagocytosis was consistently diminished in leukocytes incubated with colchicine in concentrations as low as 2.5 x 10(-6) mole per L (1 mug per ml), and this inhibition was dosage dependent. Yet there was no evidence of decreased phagocytosis with concentrations of colchicine as high as 2.5 x 10(-4) mole per L (100 mug per ml). Furthermore, with measurements at 20, 40, and 60 minutes, the rate of phagocytosis was comparable with and without colchicine.A clue to the dissociation between oxygen consumption and phagocytosis was found in rapidly dried preparations of the incubated leukocytes. Ingested bacteria were present in both control and colchicine-treated granulocytes. In addition, control cells showed normal loss of granules (lysosomal particles) and prominent cytoplasmic vacuoles (digestive vacuoles). Colchicine-treated cells, however, showed less such degranulation and vacuolization. Measurements of granule-associated acid phosphatase activity after phagocytosis support the morphologic observations of less degranulation in colchicine-treated leukocytes. The muted metabolic and morpholgic response to phagocytosis in colchicine-treated cells may be important for the anti-inflammatory effect of colchicine in acute gouty arthritis. Colchicine may also find wider use in defining structure-function dependencies in metabolically stimulated cells.


Cerebral blood flow and metabolism in the Wernicke-Korsakoff syndrome. Cerebral blood flow and metabolism were measured by the iodoantipyrine-4-(131)I method in nine patients and by the nitrous oxide method in three patients with the Wernicke-Korsakoff syndrome.Cerebral blood flow and cerebral oxygen and glucose consumption were strikingly reduced from the normal, whereas cerebral vascular resistance was increased. Total cerebral metabolism and blood flow may be greatly reduced even though the cerebral metabolic defect is confined to circumscribed anatomical areas. Profound reduction in brain metabolism is not necessarily reflected in alterations of consciousness or awareness as has been previously suggested, or in electroencephalographic abnormalities. This appears to provide cogent support for the neurophysiological principle that disturbance of consciousness is a function of the location of the lesion, not the over-all degree of metabolic defect. The absence of improvement of cerebral metabolic functions in two patients who were restudied after an additional 2 to 3 weeks of treatment confirms the clinical impression of incomplete recovery in many such patients.


Studies on the intracerebral toxicity of ammonia. Interference with cerebral energy metabolism due to excess ammonia has been postulated as a cause of hepatic encephalopathy. Furthermore, consideration of the neurologic basis of such features of hepatic encephalopathy as asterixis, decerebrate rigidity, hyperpnea, and coma suggests a malfunction of structures in the base of the brain and their cortical connections. The three major sources of intracerebral energy, adenosine triphosphate (ATP), phosphocreatine, and glucose, as well as glycogen, were assayed in brain cortex and base of rats given ammonium acetate with resultant drowsiness at 5 minutes and subsequent coma lasting at least 30 minutes. Cortical ATP and phosphocreatine remained unaltered during induction of coma. By contrast, basilar ATP, initially 1.28 +/- 0.15 mumoles per g, was unchanged at 2.5 minutes but fell by 28.1, 27.3, and 26.6% (p < 0.001) at 5, 15, and 30 minutes after NH(4)Ac. At comparable times, basilar phosphocreatine fell more strikingly by 62.2, 96, 77.1, and 71.6% (p < 0.001) from a control level of 1.02 +/- 0.38 mumoles per g. These basilar changes could not be induced by anesthesia, psychomotor stimulation, or moderate hypoxia and were not due to increased accumulation of ammonia in the base. Glucose and glycogen concentrations in both cortex and base fell significantly but comparably during development of stupor, and prevention of the cerebral glucose decline by pretreatment with glucose did not obviate ammonia-induced coma or the basilar ATP fall. These findings represent the first direct evidence that toxic doses of ammonia in vivo acutely affect cerebral energy metabolism and that this effect is preferentially localized to the base of the brain.


Correlations of mechanical stability, morphology, pulmonary surfactant, and phospholipid content in the developing lamb lung. Pressure-volume characteristics and surface tension measurements of the lamb of 120 to 130 days gestational age were typical of the mature lung in the upper lobes and the immature lung in the lower lobes. By term both upper and lower lobes had findings characteristic of the mature animal. Phospholipid concentration per milligram DNA and per cent saturated fatty acids on pulmonary phosphatidyl choline were relatively constant from 60 to 120 days gestational age; thereafter there was a significant increase in both measurements. These changes usually coincided with an increase in osmiophilic inclusion bodies in the large alveolar cell.A concentration of disaturated phosphatidyl choline per milligram DNA in excess of 0.170 mg per mg was associated with a minimal surface tension below 13 dynes per cm (p < 0.001). Newborn animal lungs contained over 3 times this critical concentration, whereas adult lungs contained 1.5 times this value. The excess disaturated phosphatidyl choline per milligram DNA may represent a reservoir of pulmonary surfactant.