Magnesium dependence of the association kinetics of Escherichia coli ribosomal subunits.

The magnesium dependence of the Escherichia coli ribosomal subunits association has been investigated by the stopped-flow technique using isolated 30S and 50S particles depleted of polyamines and any initiation factor. Binding of the fluorescent probe bis(8-anilino-1-naphthalenesulfonate) to the ribosomal proteins occurs through biphasic kinetics. A dark reaction corresponding to a very rapid, reversible complexation of the dye molecule is followed by a slow photochemical reaction that gives rise to irreversible addition of the probe. Only the 30S subparticle exhibits a magnesium-dependent conformational change from the kinetic analysis of the dark reaction. The 70S formation kinetics are limited by a conformational change of the 30S subunit if this particle is depleted of Mg2+ (1 mM Mg2+/50 mM K+), while its activated structure is restored by incubation with 8 mM Mg2+/50 mM K+. No rate-limiting conformation rearrangement of the 50S subunit could ever be evidenced. The Mg2+ dependence of the association kinetics of preactivated ribosomal particles is satisfactorily explained by electrostatic effects and/or formation of salt bridges, in agreement with the results of Wishnia and co-workers (Wishnia, A. Boussert, A., Graffe, M., Dessen, P., and Grunberg-Manago, M. (1975), J. Mol. Biol. 93, 499). Equilibrium studies indicate that the ribosomal preparations we used are of B type, according to Debey et al. (Debey, P., Hui Bon Hoa, G., Douzou, P., Godefroy-Colburn, T., Graffe, M., AND Grunberg-Manago, M. (1975) Biochemistry 14, 1553). The addition of spermidine results in a drastic fall of the need of Mg2+ for association, but it does not allow conversion of B-type particles into A-type ones at 25 degrees C. In addition to that, some 30S-bound spermidine appears to be involved directly in the coupling reaction.


Allosteric interpretation of Mg2+ binding to the denaturable Escherichia coli tRNAGlu2+.

The Mg2+ binding properties of the denaturable tRNAGlu2 from E. coli in 0.1 M Na+, pH7, are characterized by equilibrium dialysis. At 34 degrees C, where the native and denatured conformers are in equilibrium, Mg2+ binding is cooperative. By trapping the tRNA completely in the native conformation at 4 degrees C it is shown that native tRNAGlu2 possesses one strong binding site, K1 = 7.5 x 10(4) M-1 and approximately 36 weak sites with K2 = 8.3 x 10(2) M-1. A significantly lower affinity for the denatured conformer is indicated. We show that Mg2+ effects an allosteric transition from the low affinity denatured conformational state to the high affinity native state and develop the appropriate equations to fit the Mg2+ binding data with physically meaningful parameters. Our results also suggest the previously reported cooperative cation binding to tRNA arises from a cation induced conformational change to the native tRNA conformation and does not reflect the inherent Mg2+ binding properties of the native conformer.


Isolation and properties of 6-phosphogluconate dehydrogenase from Escherichia coli. Some comparisons with the thermophilic enzyme from Bacillus stearothermophilus.

6-Phosphogluconate dehydrogenase (6-phospho-D-gluconate:NADP oxidoreductase (decarboxylating), EC 1.1.1.44) of Escherichia coli MREp 600 has been isolated with the purpose of carrying out comparative studies with the thermostable enzyme previously isolated from Bacillus stearothermophilus (Veronese, F.M., Boccù, E.,Fontana, A., Benassi,C.A., and Scoffone, E. (1974), Biochim, Biophys. Acta 334, 31). The purified enzyme appeared homogeneous by the criteria of disc gel electrophoresis with and without sodium dodecyl sulfate, ultracentrifugation, and gel filtration. The enzyme has enzymological and physiochemical properties similar to the enzyme isolated from other sources, including B. stearothermophilus. The E. coli enzyme has a mol wt of 100,000 +/- 3000 and is composed of two apparently identical subunits. The amino acid composition of both the mesophilic and thermophilic enzyme has been determined and found to present large similarities. The E. coli enzyme shows a high degree of specificity for nicotinamide adenine dinucleotide (NADP) and it is inhibited by reduced NADP (NADPH). Cysteine residues are involved in the catalytic activity, since on incubation of the enzyme with p-chloromercuribenzoate or 5.5'-dithiobis(2-nitrobenzoic acid) strong inhibition occurs, activity being restored by treatment with excess of beta-mercaptoethanol. The substrate 6-phosphogluconate protects partially the enzyme from inactivation. Both the mesophilic and thermophilic 6-phosphogluconate dehydrogenases are inactivated by Rose Bengal in the presence of light by similar kinetics and protected against photoinactivation by the enzyme substrate. The E. coli enzyme, on the other hand, showed distinct differences in stability against heat and unfolding agents in respect to the B. stearothermophilus enzyme. Heating at 50 degrees C or incubation in 8 M urea results in rapid inactivation. The gross structure of the mesophilic and thermophilic enzyme was very similar as judged by circular dichroic measurements. The far-ultraviolet circular dichroic spectrum had a negative band centered at about 220 nm. In both cases, the fluorescence emission spectrum indicates that the environment of the tryptophan residues is similar, since both enzymes show an emission maximum at 334 nm upon excitation at 295 nm. Circular dichroism measured at various temperatures between 25 and 80 degrees C showed the mesophilic enzyme to be conformationally stable below about 45 degrees C and the thermophilic enzyme below 60 degrees C. The secondary structure of the E. coli enzyme was very sensitive to the denaturing action of urea, since in 8 M urea it rapidly unfolded. Partial renaturation after urea treatment occurred on dilution with buffer or dialysis, as evidenced by spectral properties of the renatured enzyme. The results show that the mesophilic and thermophilic enzymes are very similar and that differences in thermal stability depend on subtle differences in the architectures of the proteins.


Hydrolytic action of aminoacyl-tRNA synthetases from baker's yeast: "chemical proofreading" preventing acylation of tRNA(I1e) with misactivated valine.

Phe-tRNAPhe-C-C-A, Val-tRNAVal-C-C-A, and Ile-tRNAIle-C-C-A, which accept their amino acid on the 2'-OH of the 3'-terminal adenosine, are hydrolyzed readily by their aminoacyl-tRNA synthetase. If the 3'terminal adenosine in these tRNAs is replaced by either 3'-deoxyadenosine or formycin, little if any hydrolysis can be observed. Correspondingly Ser-tRNASer-C-C-A which accepts serine on the 3'-OH of the 3'-terminal adenosine is hydrolyzed by seryl-tRNA synthetase, whereas Ser-tRNASer-C-C-2'dA and Ser-tRNASer-C-C-F are not. Tyr-tRNATyr-C-C-A and all modified Tyr-tRNATyr-C-C-N, which can accept tyrosine on either the 2'OH or the 3'-OH of the 3'terminal adenosine, are not hydrolyzed by tyrosyl-tRNA synthetase. The data can be rationalized assuming that hydrolysis takes place only if the amino acid is bound to the nonaccepting OH and hence is not positioned at the amino acid binding site upon formation of the complex between aminoacyl-tRNA and aminoacyl-tRNA synthetase. In the formycin-carrying tRNA, the amino acid bound to the nonaccepting OH seems to be inaccessible to the enzymatic groups responsible for hydrolysis. Val-tRNAIle-C-C-3'dA and Ile-tRNAIle-C-C-3'DA cannot be hydrolyzed by isoleucyl-tRNA synthetase. Val-tRNAIle-C-C-A is hydrolyzed by the enzyme five times more rapidly than Ile-tRNAIle-C-C-A. Whereas Ile-tRNAIle-C-C-F is absolutely stable, Val-tRNAIle-C-C-F si hydrolyzed immediately. As shown by the earlier finding that valine misactivated by isoleucyl-tRNA synthetase cannot be permanently transferred to tRNAIle-C-C-A but to tRNAIle-C-C-3'dA, the 3'-OH is essential for preventing transfer of misactivated valine. It thus appears that valine is hydrolyzed off Val-tRNAIle-C-C-N if it is bound to the accepting 2'-OH in the binding site for isoleucine. A hypothesis is offered attempting to explain the experimental observations in mechanistic terms. We consider the hydrolytic action of the aminoacyl-tRNA synthetases as a general mechanism of "chemical proofreading" in the protein biosynthesis.


Reductive microbial conversion of anthracycline antibiotics.

Reductive conversion of several anthracycline glycosides to their 7-deoxyaglycones occurs during their microaerophilic incubation with strains of Aeromonas hydrophila, Citrobacter freundii, and Escherichia coli. Further, extracts of microaerophilically grown A. hydrophilia catalyze DPNH-dependent reductive conversion of the same compounds. Anthracycline substrates cleaved by both whole cells and by the cell-free system include steffimycin, steffimycin B, nogalamycin, cinerubin A, and daunomycin. Investigation of glycoside cleavage as a function of both time and anthracycline concentration demonstrated the superiority of A. hydrophila over C. freundii and E. coli in regard to reaction rate and efficiency of conversion. Interestingly, some degree of anaerobicity was required for glycoside cleavage by all three organisms. Evidence supporting 7-deoxyaglycone formation via direct reductive cleavage, as opposed to a multienzyme-catalyzed process involving hydrolysis followed by dehydration and reduction, includes the following. Equilibrium mixtures of glycoside substrate and 7-deoxyaglycone product prepared using both whole cells and their extracts display no anthracycline hydrolysis products. Further, authentic steffimycinone (aglycone), the expected product of hydrolytic sugar cleavage of steffimycin, was shown to be converted to 7-deoxysteffimycinone (7-deoxyaglycone) at a rate slower than steffimycin. These data indicate that, if steffimycinone were present as an unbound metabolic intermediate, it should have been visible in the equilibrium mixture, but none was detected.


Purification of thymidylate synthetase from enzyme-poor sources by affinity chromatography.

The adsorption of thymidylate synthetase from Escherichia coli B to aminoalkyl-Sepharose with the increasing length of carbon chain (2--6 carbon atoms) was investigated. A correlation was found between the chain length and adsorption effectiveness, increasing from the two- to the six-carbon chain. A hydrophobic chromatography of the enzyme on aminobutyl-Sepharose gave about 20-fold purification. A new affinity chromatography carrier was synthesized containing tetrahydromethotrexate linked to aminoethyl-Sepharose via its carboxylic groups. The carrier adsorbed the enzyme from the crude preparation only in the presence of deoxyuridine 5'-monophosphate (dUMP) in a concentration of 2 X 10(-5) M. The specifically adsorbed thymidylate synthetase was eluted with sacharose-containing buffers in which dUMP was omitted. The purification procedure was applied to a crude thymidylate synthetase preparation from resting E. coli, calf thymus, Sarcoma 180, and Gardner lymphosarcoma. The purified enzyme from all mentioned sources showed one protein band on disc electrophoresis corresponding to enzymatic activity. The formation of a reversible noncovalent complex enzyme-tetrahydromethotrexate-dUMP on the affinity column is supposed.


Circular dichroism study of the interaction of glutamyl-tRNA synthetase with tRNAGlu2.

The interaction of glutamyl-tRNA synthetase with tRNAGlu2 has been studied. The enzyme was purified to apparent homogeneity, and consists of a single chain with a molecular weight of 59 000. The sedimentation coefficient (sdegrees20,w) was found to be 3.7 S and suggests this enzyme is quite asymmetric. The enzyme binds 1 mol of tRNAGlu2 and has a binding constant of 5 X 10(6) M-1 at pH 7.0 in 0.1 M sodium chloride. A circular dichroic study of the interaction under the same solvent conditions implied both the synthetase and tRNAGlu2 underwent a change in conformation as the complex was formed. In the case of the enzyme there appears to be some loss of alpha-helical structure. The tRNAGlu2 results can be interpreted to indicate a change in the conformation of one or more of the helical regions of this molecule. A residue in the anticodon loop, 5-methylaminomethyl-2-thiouridine, has a distinct circular dichroic band at 340 nm in the free tRNAGlu2. As the complex is formed this band is shifted to the blue. This was interpreted to indicate that the enzyme forms a hydrogen bond with this residue in the anticodon loop, with a change in the conformation of the loop possibly also having occured.


Energy metabolism of Saccharomyces cerevisiae discrepancy between ATP balance and known metabolic functions.

The contribution of metabolic pathways to the catabolism of glucose, galactose and ethanol by Saccharomyces cerevisiae in aerobiosis has been studied. The results suggest that: 1. Of the total ATP formed in catabolism yeast obtain as much as 60% from ethylic fermentation during logarithmic growth on glucose. However, about 80% of ATP is formed in oxidation of galactose. Oxidation seems to be the only important catabolic pathway of ethanol. 2. The ratios between growth yield and ATP formed in catabolism were approx. 9, 7 and 3 g dry yeast/mol ATP in glucose, galactose and ethanol cultures, respectively. 3. The balance between ATP produced in catabolism of substrates and the requirements of ATP for the biosynthesis of cellular material indicates that as much as 60% of ATP is spent in functions other than net biosynthesis. 4. The rate of ATP expenditure in non net-biosynthetic functions during growth was approx. 20 mmol/g dry yeast per h. 5. In conditions in which no growth occurred but cell viability was maintained, that is, in the absence of exogenous carbon and nitrogen source, the ATP production rate was approx. 1 mmol ATP/g dry yeast per h. 6. These results indicate that the ATP required for maintaining the yeast alive, what would be considered maintenance energy "sensu stricto", is only a minor proportion of the ATP spent in non net-biosynthetic functions during growth. The identification of the processes related to growth which spend more energy than that required for net biosynthesis could lead to important insights in cell biology.


ATP concentration in Escherichia coli during oxygen toxicity.

Escherichia coli, strain E-26, grown in defined salts medium with glucose as the sole carbon and energy source, contained 1.50+/-0.16-10(6) molecules of ATP/cell. ATP was extracted with HC104 and assayed with a Dupont Luminescence Biometer using the luciferin-luciferase assay. Exposure during exponential growth at 37degreesC to 4.2 atm of oxygen resulted in complete growth cessation within 5 min, and to cyclic changes in cellular ATP concentration over a 2 h period. However, significant decrease in cellular ATP concentration occurred after growth inhibition in hyperbaric oxygen; hence, lack of ATP was not the cause of growth inhibition from oxygen toxicity.


The isolation and properties of a DNA-directed RNA polymerase from yeast mitochondria.

A method is described for the rapid isolation of yeast mitochondrial DNA-directed RNA polymerase. The enzyme obtained had a specific activity of 1.56 nmol UMP incorporated per mg protein in 20 min at 37 degrees C, and is some 95% pure. This purified enzyme upon  polyacrylamide gel elctrophoresis consists of a single polypeptide of 68 000 mol. wt. However, the enzyme forms aggregates easily which are affected by ionic strength, an increase decreasing the apparent molecular weight of the aggregates. This property also explains the presence of two peaks of activity upon DEAE-cellulose chromatography. The purified enzyme is still sensitive to rifamycin and to a number of rifamycin derivatives. The enzyme's sensitivity to rifamycin and rifamycin derivatives was compared with Escherichia coli and yeast nuclear RNA polymerases.


Spermine stabilizes the conformation of tRNAPhe in crystals.

Crystals from yeast tRNAPhe were dissolved and compared with tRNAPhe that had not been srystallized. A number of differences were found regarding the interaction with ethidium bromide, the melting point and the circular dichroic signal. These differences were assigned to the presence of spermine in the dissolved crystals indicating a transient stabilization of the conformation of tRNAPhe, probably as a tRNAPhe-spermine complex, after dissolving.


Permeability to cobalt ions and action of colicin K in a mutant that overproduces cardiolipin.

A mutant of Escherichia coli that produces excess cardiolipin becomes less capable of transporting Co2+. Cardiolipin therefore does not act as an ionophore under these conditions. Colicin K brings about the typical increase in permeability to Co2+ in the mutant.


Effect of oral papaverine on cerebral blood flow in normals: evaluation by the xenon-133 inhalation method.

Several issues related to the use of vasoactive drugs in the treatment of neuropsychiatric disorders secondary to cerebrovascular insufficiency remain unresolved due to lack of a practical method for the quantitative measurement of cerebral blood flow (CBF). The present report concerns the use of a noninvasive CBF method in the evaluation of a commonly used vasoactive drug (oral papaverine). Series CBF measurements were made by the Xe133 inhalation method in 21 healthy young adults, using a double-blind crossover experimental design. The findings show that oral papaverine (300 mg b.i.d.) significantly increases blood flow under conditions of both normal breathing and hyperventilation, the latter being used to induce cerebral vasoconstriction. There were large variations in individual response to the drug, and the average increase in CBF (6 to 9%) was smaller than that observed by other investigators using intravenous papaverine. No adverse reactions or alterations in blood pressure were encountered.


Urinary excretion of 3-methoxy-4-hydroxyphenylglycol in depressed patients: modifications by amphetamine and lithium.

The excretion of 3-methoxy-4-hydroxyphenylglycol (MHPG), the urinary metabolite which may best reflect brain norepinephrine, was measured in eight depressed patients on a longitudinal basis during the administration of placebo, d- and 1-amphetamine (30 mg/day), lithium carbonate (0.9 - 2.1 g/day), and d- and 1-amphetamine together with lithium. d-Amphetamine significantly decreased MHPG excretion. No significant change in MHPG was observed during administration of 1-amphetamine or when d- or 1-amphetamine was given together with lithium carbonate. Six patients responded with behavioral activation and euphoria to d-amphetamine, and these patients tended to have lower base line MHPG values in comparison to the nonresponders. Reductions in MHPG excretion during amphetamine administration were greatest in the patients with minimal behavioral responses to the drug, while some of the patients demonstrating more marked stimulant effects had elevated levels of MHPG during the amphetamine administration period. Variable changes, including elevated levels of MHPG in the responders, might represent the contribution of amphetamine-related changes in clinical state, including increased physical activity and (in two of the responding patients) the development of hypomania.


Two cases of malaria presenting with psychiatric symptoms.

Mood and behavior disorder may appear either due to primary psychiatric disorder or may signify an underlying, frequently undetected, physical pathology. Two instances of malaria falciparum presenting with psychosocial symptomatology are described.


A psychometric study of chronic cerebellar stimulation in man.

Groups of patients with intractable epilepsy or spasticity of varying etiologies underwent psychological testing before and during chronic cerebellar cortex stimulation, a neurosurgical technique designed for the relief of these symptoms. The battery of psychological tests permitted a standardized assessment of intellectual, memory, and perceptual functions. Other behavioral dimensions were assessed through structured interviews. No apparent declines in higher integrative functions followed shorter or longer term cerebellar stimulation. In contrast, stimulated patients tended to show increments in tests of recent memory and verbal output beyond that of unoperated comparison groups. Subjective reports of increased "alertness" and reduced depression and anxiety were also frequently given. Psychological and neural factors may both contribute to the observed behavioral alterations. It was concluded that the cerebellum participates in behavioral functions by modifying cortical and subcortical mechanisms relevant to integrative behavior and emotions. Specific hypotheses were presented.


Distribution of transplantation antigens on cell surfaces.

Significant advances have been made over the past few years in elucidating the genetics, the chemical composition, and, more recently, the in situ relation of the major histocompatibility antigens of the mouse and man. Attempts to map the arrangement of individual antigens on the surface of cells have revealed that some antigens specified by a given subregion of both the H-2 and HL-A systems are in close proximity on the cell membrane and that attachment of antibody to one site to a certain degree blocks or inhibits the binding of antibody to the adjacent site. Allelic antigens in the H-2 system tend to inhibit binding. H-2D and H-2K antigens show either inhibition or noninteraction, possibly reflecting a cis-trans effect. Unlike with the H-2, inhibition of binding occurs only between HL-A antigens specified by homologous chromosomes. Also, a number of instances have been noted where inhibition of binding is unidirectional, possibly reflecting a polymeric nature of antigen or stratification of moieties at cell surface. Inhibition of antibody attachment between several alloantigenic systems on thymocytes in mice and also a variation in the mobility of the histocompatibility antigens suggest that the moieties bearing histocompatibility antigens are comprised of several gene products. Further work is needed to establish the validity of this assumption and to fully define the composition of these units. Ample data have been obtained from both biological and biophysical experiments to support the suggestion that single or multiple complexes of glycoproteins can move in the plane of the membrane. Although the composition of these cell membrane components remains a question, direct visualization by fluorescence and electron microscopy indicates that these moieties are small and, under natural conditions, distributed uniformly over the cell surface. Direct and indirect labeling techniques have shown that the complexes have no fixed position in the cell membrane and can be displaced laterally in the plane of the membrane without affecting the distribution of other surface molecules, such as sIg and species-specific antigens. Additional evidence suggests that H-2D and H-2K antigen complexes, as well as their gene products specified by different parent chromosomes, may be displaced separately. These observations are especially interesting and must be reconciled and data obtained by proximity analysis which indicate an association of some allelic products and possibly certain combinations of D and K antigens. Whether the differences noted in reactivity of the various surface antigens following attachment of antibodies are attributable to difference in size or to differences in the manner of their intercalation in the cell membrane remains to be elucidated. The rapid advances in elucidation of the molecular structure of biological membranes suggest that experimental work should be done on the biophysics of the structure of the antigenic sites and the mechanism of migration...


Intracellular localization and immunogenic capacities of phenotypic products of mouse histocompatibility genes.

The transplantation antigens are the phenotypic products of genes which control histocompatibility in vertebrate species. The products of major histocompatibility locus of the mouse, H-2, have been studied as a model. The H-2 transplantation antigens are expressed on cellular membranes in all tissues examined. These gene products have been isolated from cells associated with subcellular membranes. These membranes have been assayed both for their antigen content (antigenicity) and for their capacities to induce a primary humoral and a cell-mediated response (immunogenicity). In all tissues examined, the H-2 antigens (products of the K and D regions of H-2) were found expressed in high concentration on cell surface membrane. However, immunogenic activity was observed only with spleen and thymus preparations, consisting mainly of intracellular membranes (MLP). Immunogenic MLP was also isolated from lymphoblast and fibroblast cells, and again was derived mainly from endoplasmic reticulum. In other tissues, such as liver, kidney, and erythrocytes, H-2 antigens were found only on surface membrane and in an antigenic but nonimmunogenic form. A novel method for tagging surface membrane of mammalian cells is presented. It consists of binding, to whole cells in a covalent linkage, purified preparations of the beta-galactosidase of E. coli. The bound enzyme has proved to be an unambiguous marker for surface membrane. With this marker, the stability of surface membrane to shear forces during homogenization could be assessed. A number of considerations suggest that immunogenicity of transplantation antigens may be due to factor(s) present on the membranes in addition to the H-2 antigenic determinants. There are indications that these factors may be controlled by the I region of the H-2 complex. It is interesting to note that normal tissues which have Ia antigens on their surface membranes yield immunogenic MLP (spleen and thymus), whereas those without Ia surface antigens yield an antigenic MLP that has no immunogenic capacity (liver, kidney, and erythrocytes).


The lagged normal family of probability density functions applied to indicator-dilution curves.

The generalized lagged normal functions are a family of probability density functions whose members consist of a normal density function convolved with one or more exponential functions. The functions are not expressible in closed form, but an error function approximation procedure allows rapid and accurate calculation of the value of these functions. The approximation procedure can be used in conjunction with a nonlinear regression program to fit a lagged normal function to experimental data.


Decrease of endothelaemia during immunosuppression.

The counts of circulating endothelial cells estimated by a new quantitative method were significantly decreased in a group of patients after renal transplantation. Subsequent animal studies have shown that such a decrease may be attained by the administration of immunosuppressive drugs. The explanation of this observation may be based on the suppression of a normal endothelial cell replacement.


Casein phenotyping in witch's milk.

18 neonatal colostrum samples were analyzed by urea-starch-gel electrophoresis to determine their casein patterns. It was found that precise typing of both alpha-and beta-casein could be obtained after the secretory burst. This genetic marker can be employed at the family level.


Simultaneous allotransplantation of kidney and spleen in dogs with or without previous splenectomy.

29 simultaneous kidney and spleen transplantations have been performed on dogs. Time of rejection, similar for both organs, occurs in a delay very close to the control series. No statistically difference of the day of rejection was observed in relation with the implantation of the spleen either on the iliac or on the mesenteric vessels. Splenectomy few weeks prior to transplantation had no effect on the day and the histological type of rejection of both transplanted organs.


Genetic resistance to marrow transplantation as a leukemia defense mechanism.

(C57 X AKR) F1 hybrid mice show genetic resistance to C57 parental bone marrow cells, but not to AKR parental bone marrow cells. (C3H X AKR) F1 hybrids show no genetic resistance to bone marrow transplantation from either parental strain. Transplantation of AKR lymphoma cells into lethally irradiated "resistant" (C57 X AKR) F1 and "non-resistant" (C3H X AKR) F1 hybrids produce lymphomatous spleen colonies in "non-resistant" hybrids but not in "resistant" hybrids. Thus "resistant" (C57 X AKR) F1 hybrids can recognize and reject AKR lymphoma cells, but not normal AKR bone marrow cells. A normal biological role of lymphoma surveillance is postulated for genetic resistance.


Capillary migration of the cells of three murine tumours.

Cells of a murine myeloid leukemia, a lymphoid leukaemia, and a reticulosarcoma, were tested for their ability to migrate from capillary tubes. As compared with normal cells from the speeen, bone marrow, lymph nodes or peritoneal exudate, tumour cells migrated rather poorly. When present, the migration could be attributed to the existence of normal cells in the suspension of tumour cells. No correlation has been found between the migratory behaviour and the mode of growth of the tumour "in vivo".


Mobility measurement by analysis of fluorescence photobleaching recovery kinetics.

Fluorescence photobleaching recovery (FPR) denotes a method for measuring two-dimensional lateral mobility of fluorescent particles, for example, the motion of fluorescently labeled molecules in approximately 10 mum2 regions of a single cell surface. A small spot on the fluorescent surface is photobleached by a brief exposure to an intense focused laser beam, and the subsequent recovery of the fluorescence is monitored by the same, but attenuated, laser beam. Recovery occurs by replenishment of intact fluorophore in the bleached spot by lateral transport from the surrounding surface. We present the theoretical basis and some practical guidelines for simple, rigorous analysis of FPR experiments. Information obtainable from FPR experiments includes: (a) identification of transport process type, i.e. the admixture of random diffusion and uniform directed flow; (b) determination of the absolute mobility coefficient, i.e. the diffusion constant and/or flow velocity; and (c) the fraction of total fluorophore which is mobile. To illustrate the experimental method and to verify the theory for diffusion, we describe some model experiments on aqueous solutions of rhodamine 6G.


Geometry of normal mammalian platelets by quantitative microscopic studies.

The shape distributions of normal and hardened human and rabbit erythrocytes and platelets were obtained for edge-on orientations of a few hundred freely rotating cells from analyses of microphotographs obtained similarly as by Ponder(1930, Q. J. Exp. Physiol. 20:29) by phase-contrast microscopy at 800 X magnification. Major average diameters (d) and thicknesses (t) were estimated for both normal and hardened cells, and were used to calculate an average geometric axis ratio, rp = t/d, which increases to unity as cells become more spherical. Our fixation procedure did not alter these shape parameters: rp was unchanged for erythrocytes, with d and t values similar to those reported by Ponder (1930); platelets had d X t = 3.6 +/- 0.7 mum X 0.9 +/- 0.3 mum and 3.1 +/- 0.4 mum X 0.6 +/- 0.3 mum, respectively, for human and rabbit cells, with rp = 0.26 and 0.20, respectively. Agreement in rp was found with data obtained by a novel rheo-optical method which allows for a direct statistical averaging for large populations (greater than 100 X 10(3) cells). Histograms and linear correlation studies were made of the above three parameters (d,t,rp), as well as volume (V), total surface area are (S), and sphericity index (S.I.) calculated for both "prolate ellipsoid" and "disc with rounded edges" models. Results indicate very high linear correlations between rp - t, rp - S. I., and d -S, with high correlations for t - V,d -V and S. Data are in agreement with the few reports in the literature determined by other methods, with the best model for platelets appearing to be an oblate spheroid.


The so20,w of unsheared DNA from whole cell lysates of Escherichia coli.

We present measurements of the sedimentation coefficients of DNA present in whole cell lysates of E. coli. The method used is a preparative version of the band sedimentation experiment of Bruner and Vinograd. We show that in order to obtain reliable data on the time dependence of sedimentation, it is necessary to accelerate and decelerate the rotor over much longer times than the standard centrifuge allows. We describe the necessary modifications to the preparative centrifuge and use them to determine the So20,W of unsheared E. coli DNA. The value for the fastest moving components in the lysate is 220 S. The molecular weight of the DNA corresponding to this sedimentation coefficient is probably 1.7 X 10(9) g/mole. However, alternative values cannot be ruled out.


Separation of tumor cells by density gradient centrifugation: recent work with human tumors and a discussion of the kind of quantitation needed in cell separation experiments.

The application of centrifugation to the purification of individual kinds of cells from tumors has been reviewed. Recent applications of a previously described isokinetic gradient to the purification of subpopulations of cells from human tumors are presented. Individual kinds of cells have been separated from Hodgkin's disease by a method described previously, and the cells from Hodgkin's disease which contain the tumor associated antigen described by Order and his colleagues have been identified for the first time. The importance of the thorough documentation of cell separation procedures is discussed, and we have specifically listed several kinds of data which we believe are important in the documentation of cell separation techniques.


Carbon dioxide inhibition of yeast growth in biomass production.

Saccharomyces cerevisiae was grown under aerobic and substrate-limiting conditions for efficient biomass production. Under these conditions, where the sugar substrate was fed incrementally, the growth pattern of the yeast cells was found to be uniform, as indicated by a constant respiratory quotient during the entire growing period. The effect of carbon dioxide was investigated by replacing portions of the nitrogen in the air stream with carbon dioxide, while maintaining the oxygen content at the normal 20% level, so that identical oxygen transfer rate and atmospheric pressure were maintained for all experiments with different partial pressures of carbon dioxide. Inhibition of yeast growth was negligible below 20% CO2 in the aeration mixture. Slight inhibition was noted at the 40% CO2 level and significant inhibition was noted above the 50% CO2 level, corresponding to 1.6 X 10(-2)M of dissolved CO2 in the fermentor broth. High carbon dioxide content in the gas phase also inhibited the fermentation activity of baker's yeast.


Utilization of cellulosic materials through enzyamtic hydrolysis. I. Fermentation of hydrolysate to ethanol and single-cell protein.

Ethanol fermentation studies were conducted with Saccharomyces cerevisiae ATCC "4126, to determine the optimal conditions of oxygen tension and feed sugar concentration. In long-term continuous culture maximum ethanol production was found to occur at 0.07 mmHg oxygen tension and 10% glucose feed concentration. Preliminary process design and cost studies are developed for industrial scale fermentations to produce ethanol and torula yeast from sugars obtained by enzymatic hydrolysis of newsprint.


Allogeneic marrow grafting for treatment of aplastic anemia: a follow-up on long-term survivors.

Eleven of twenty-four patients with severe aplastic anemia given marrow grafts from HLA-identical siblings between October 1970 and March 1973 are alive with normal marrow function and continued evidence of engraftment 3-5 yr later. Ten have been leading normal lives with no immunosuppressive or other drug therapy since day 100 postgrafting. One has had chronic graft-versus-host disease of the skin which is now slowly improving with no therapy. He returned to full-time employment in the summer of 1975. The long-term well-being of almost half of our initial patients emphasizes the importance of marrow transplantation for the treatment of severe aplastic anemia.


Establishment of erythropoiesis following bone marrow transplantation in a patient with congenital hypoplastic anemia (Diamond-Blackfan syndrome).

Marrow transplantation was attempted in a 13-yr-old boy with congenital hypoplastic anemia who had never responded to corticosteroid therapy. Prior to the transplant, he had received 238 transfusions, at least 12 of which were from his father. He was prepared for grafting with antilymphocyte globulin, procarbazine, and total body irradiation (1000 rads). The patient, whose red cells were Group B, then received marrow cells from his Group O, histocompatible, sister. Thereafter, reticulocytes, Group O erythrocytes, and female leukocytes appeared in the peripheral blood. Erythroid precursors were seen in the patient's marrow for the first time in his life, and all lacked fluorescent Y chromosomes. Dividing cells were all female. After initially progressing well, the patient developed interstitial pneumonia and died 55 days after the transplant. The successful erythroid graft suggested that this patient's failure to produce red blood cells was due to a defective stem cell rather than to a humoral defect, plasma inhibitor, or abnormal marrow microenvironment. It suggested further that sibling marrow may be engrafted in patients who have received multiple transfusions, even from a parent.


Effect of sodium valproate on tardive dyskinesia.

The effect of sodium valproate, a drug which has been demonstrated to increase gamma-aminobutyric acid levels in the CNS, on tardive dyskinesia and psychiatric symptoms was investigated in a double-blind cross-over study on 32 chronic psychiatric patients. The oro-facial dyskinesias were totally or significantly relieved in 17 cases. During the active treatment period the involuntary movements of the extremities and dystonic spasms were also significantly relieved in 7 out of 9 patients. In two patients, however, the extrapyramidal symptoms became slightly worse. A significant improvement was noted in the psychiatric symptoms of 14 out of 32 patients during sodium valproate administration. The psychiatric state of 4 out of 32 patients deteriorated. Three was no correlation between the serum concentration of sodium valproate and its effect on the dyskinesia or on the psychiatric symptoms. Some of the elderly subjects showed a slight accumulation of the drug.


A dialogue with Mandeville.

In A Treatise of the Hypochondriack and Hysterick Passions published in 1711, Bernard Mandeville, writing by way of Information of Patients' describes three dialogues between an imaginary physician, Philoperio, and his patient, a confirmed hypochondriac, Misomedion. The book is original in its conception inasmuch as it represents an early recorded conversation between a physician and his patient. As such it represents a development in the writing on therapeutic technique of the eighteenth century. Though Mandeville's originality has been emphasized by contemporary medical historians, this book seems to have had little influence on the therapeutic style of the physicians of his day.


The origins and practice of psychodrama.

Psychodrama, a method of human relations training and psychotherapy, is traced from its creation by Moreno fifty years ago to its present resurgence with the growth of 'humanistic' psychology. The basic techniques are described, together with Moreno's concepts of social interaction, e.g. encounter, spontaneity, the moment, cathersis and role playing. Its uses and limitations are discussed briefly.


Disulfiram implantation: placebo, psychological deterrent, and pharmacological deterrent effects.

In an effort to examine the placebo, psychological deterrent, and pharmacological deterrent effects associated with implanted disulfiram, subjects were given either disulfiram implants or sham operations. Ethanol challenges elicited no disulfiram-ethanol reactions (DERs), indicating that at the time of the challenge neither a pharmacological deterrent nor a placebo effect was operating. Of the patients who resumed drinking, only those with disulfiram implants experienced DERs. Sham operation subjects continued to drink after their first post-challenge drink; four of five disulfiram implant recidivists remained abstinent following their experience of a DER. It is concluded that the pharmacological deterrent effect of the disulfiram implant may have been underestimated in previous reports.


A comparative study of flurbiprofen and aspirin in soft tissue trauma.

A double blind study using flurbiprofen (2-(2-fluoro-4-biphenylyl propionic acid) 150 mg daily and soluble aspirin (3.6 g daily) for 5 days immediately after injury, was carried out in 52 soft tissue injuries to the lower limb in professional sportsmen. Flurbiprofen was more effective than aspirin in producing analgesia (when daily pain scores were considered) after day 2 (p less than 0.02); and flurbiprofen produced a more effective resolution of soft tissue trauma when days to training and match play were considered (p less than 0.05). The inhibitory effects of flurbiprofen on prostaglandin biosynthesis and tissue action are mentioned and the use of anti-inflammatory agents given immediately after soft tissue injuries discussed.


New methods of imaging in diagnostic radiology Sylvanus Thompson Memorial Lecture.

A brief history of the recent developments in computer-assisted tomography (CAT) is presented. The development of whole-body scanners from the basic brain scanner, the trend towards higher-speed scanners using multiple detectors in a fan-type geometry, and some of the unsolved problems related to CAT scanners are discussed, as well as the potential use of whole-body scanning in radiotherapy treatment planning and in radiobiology. A review of recent developments in electrostatic imaging is presented and a new method of imaging called ionography is described in detail. It is shown how a single exposure can be used to produce a number of copies, each with a different amount of edge contrast, under the control of the radiologist. Further, it is shown how this can be done using a closed system so that the ionography chamber never needs to be opened. Some of the recent commercial developments in this field are described. It is suggested that in the future radiology departments may be replaced by departments of imaging, which will include all ways of obtaining diagnostic information and where such a department would be problem oriented rather than technique oriented. The need for basic scientific support by a medical physics and engineering group in such a department is emphasized.


Autotransplantation of the kidney for fibromuscular dysplasia of the renal arteries.

A young man was discovered to have symptomless hypertension, which subsequently proved to be caused by fibromuscular dysplasia of the renal arteries. Arterial disease was present on both sides but was more severe on the right. Samples of blood were obtained by selective catheterisation of each renal vein for the determination of plasma renin activity. The renal venous renin was greater on the right than on the left (ratio greater than 1-5 to 1) and secretion of renin by the left kidney was suppressed. Autotransplantation of the right kidney to the left internal iliac artery lowered the blood pressure at once and it has remained normal for 6 months. The implications of this procedure for the management of a bilateral disorder of the renal arteries, such as fibromuscular dysplasia, are discussed.


Amputation for vascular disease.

Amputation for peripheral ischaemia still has a depressingly high early and late mortality, and morbidity and the end result are usually less than satisfactory. Individual surgeons probably see too few amputees to treat them with maximal efficiency, and these patients create a large burden on beds and resources. There is room for improvement in all aspects of our management of amputees. Primary healing rates might be better with less heroic attempts to obtain a distal amputation. Sepsis is lessened by the use of prophylactic antibiotics. Tight bandaging and the intra-operative fitting of prostheses are undesirable. Simple tests of skin blood pressure may aid prediction of the degree of ischaemia at the proposed level of limb section and the chances of healing. The late mortality is high and merits study of methods designed to reduce it such as long term anticoagulation.


Randomised controlled trial of antibiotics in patients with cough and purulent sputum.

Two hundred and twelve adults with cough and purulent sputum of up to one week's duration were allocated randomly to treatment with doxycycline or placebo capsules for up to 10 days. Cough, purulent sputum, feeling "off colour," and time off work lasted as long in treatment and control groups, but running nose persisted for a shorter time in the doxycycline group. The number of new episodes of lower respiratory tract infections, vaginal infections, gastrointestinal upsets, and otitis media over the next six months were the same in both groups, but fewer new upper respiratory infections were experienced by the doxycycline-treated patients. There is no consensus among doctors about using antibiotics in patients with cough and purulent sputum, and these results indicate that otherwise healthy people with these symptoms will usually get better without antibiotic treatment.


Hyposensitisation with house dust mite vaccine in bronchial asthma.

In a double-blind placebo-controlled trial of tyrosine-absorbed Dermatophagoides pteronyssinus vaccine no improvement was seen in 45 patients with asthma sensitive to house dust mite. In particular there was no significant improvement in symptom scores, spirometry, and skin and nasal challenge test results. Some patients in other studies have benefited from doses of vaccine much stronger than those now commercially available, but the incidence of side effects has also been high.


Bleeding in renal failure: a possible cause.

Increased concentrations of factor VIII-related antigen (VIIIRA), factor VIII-procoagulant activity (VIIC), and decreased factor VII-von Willebrand activity (VIIIVWF) were found in the plasma of patients with chronic renal failure (CRF). This functional abnormality of the factor VII protein may partly explain the prolonged bleeding time commonly found in CRF. It was not improved by dialysis, but it was no longer found in patients with normally functioning grafted kidneys after the sixth month after transplantation. VIIIVWF levels remained decreased when compared with VIIIRA or VIIIC in transplanted patients undergoing acute reversible rejection soon after transplantation. Yet, not only VIIIC and VIIIRA but also VIIIVWF were greatly increased in patients with hyperacute irreversible rejection. Possibly a high VIIIVWF level in these patients is a thrombogenic factor.


Computer interrogation of patients.

A system of routine interrogation of patients using a computer has been developed. It includes a visual display unit with a specially designed response keyboard, and the program has been designed to adapt to the individual patient. The system was evaluated objectively, using the criteria of accuracy in eliciting symptoms, acceptability to the patient, and cost. While doctors will always take the ultimate management decisions, it seems that machines can be programmed to undertake the routine interrogation of patients, elicit evidence accurately and acceptably, and calculate the probabilities of disease as effectively as doctors.


IgM-FTA test in syphilis in adults. Its relation to clinical findings.

IgM-FTA tests have been carried out on 209 sera from 169 patients with treated or untreated syphilis at various stages and on 128 sera from 109 patients, born in areas where yaws is or was prevalent, with treated or untreated latent treponemal disease. IgM anti-treponemal antibody was found in virtually all cases of untreated early or early latent syphilis but in only 23 per cent. of sera from patients with untreated late latent syphilis. After treatment the antibody usually disappeared within one year, but it persisted in a minority of patients, including some treated for late symptomatic or congenital syphilis. Except in isolated cases there was no clinical evidence to suggest continued disease activity, although a third of the patient in whom the antibody persisted for more than 2 years after treatment were noted to be homosexuals. The test may assist in differentiating untreated early latent from late latent syphilis.


Reactivity of lymphocytes from patients with syphilis towards T. pallidum antigen in the leucocyte migration and lymphocyte transformation tests.

The reactivity of lymphocytes to Treponema pallidum antigen was studied before and after treatment in nine patients with early syphilis using a leucocyte migration test and a lymphocyte transformation test. Lymphocyte reactivity was also investigated in six patients treated for syphilis within the last 4 years, and in five untreated patients with a positive result to the T. pallidum immobilization test, but negative results to other serum tests for syphilis antibodies and without any known exposure to risk of infection by syphilis. Ten seronegative patients with different dermatological disorders served as a control group. A significant increase in lymphocyte reactivity to T. pallidum antigen was recorded in both tests in vitro after treatment. There was no difference in lymphocyte reactivity to T. pallidum antigen between the other patients studied and the control group. In early syphilis the spontaneous migration was found to be inhibited before treatment. Tuberculin skin tests were also performed and found to be suppressed in patients with primary and secondary syphilis. No difference in phytohaemagglutinin response was found between any of the groups. Plasma from patients with primary and secondary syphilis was found to change the in vitro reactivity of normal lymphocytes when stimulated with different mitogens.


Improved transport and culture system for the rapid diagnosis of gonorrhoea.

The Jembec/Neigon transport-and-culture system for the rapid diagnosis of gonorrhoea has been assessed by 167 sets of investigations in 128 named female contacts of infected men. The system offers the same advantages of efficiency and speed as the Transgrow transport-and-culture system, but it is also more flexible and much less time-consuming.


Non-specific urethritis. A placebo-controlled trial of minocycline in conjunction with laboratory investigations.

The results of a double-blind therapeutic trial on 81 men suffering from non-specific urethritis (NSU) show that minocycline was more effective than a placebo. Before treatment Chlamydia trachomatis was isolated from 31 per cent. of the men, ureaplasmas from 58 per cent., and Mycoplasma hominis from 7-5 per cent. There is evidence that chlamydiae are a cause of urethritis. However, after minocycline therapy improvement in the clinical response of patients from whom only ureaplasmas were isolated was less significant, so that the evidence that these organisms are pathogenic is less convincing. Possible reasons for this are discussed, as are the implications of finding minocycline-resistant ureaplasmas in at least 6 per cent. of the patients who harboured these organisms. The symptoms and signs of patients from whom micro-organisms were not isolated also improved after minocycline therapy, implying that the aetiological agents in this group of patients are antibiotic-sensitive. However, the results do not exclude the possibility that a minocycline-resistant agent is the cause of urethritis in a minority of patients.


Neutron scattering measurements with the label triangulation method on the 50 S subunit of E. coli ribosomes.

In the E. coli 50 S ribosomal subunit, proteins L7/L12 and L10 were deuterated by partial reconstitution. The distance between L7/L12 and L10 was measured by the label triangulation method and was found to be approximately 100 A or, with low probability, 60 to 70 A, depending on the concentration.


The production of deuterated E. coli.

Our results indicate that the preparation of deuterated macromolecules for neutron scattering from E. coli can be done easily and relatively cheaply. The entry of deuterium into macromolecules of differing species during cell growth can be controlled by varying the D2O content of the medium and the choice of carbon sources, all in minimal medium. Considering the range of deuterations obtainable with simple protonated carbon sources, there seems to be no reason to use deuterated carbon sources except for the rare occasions when a fully deuterated specimen is required.


The conversion of leucine to alpha-ketoisocaproic acid and its metabolic consequences for Escherichia coli K12.

The amino acid L-leucine serves as a good auxiliary nitrogen source for Escherichia coli K12, and in so doing is converted to alpha-ketoisocaproic acid which is excreted into the medium. L-Leucine does not serve as sole nitrogen source. Cells incubated with L-leucine as sole nitrogen source do not grow, although they do metabolize leucine, and accumulate ketoisocaproic acid in the medium. Where glycine is the only other nitrogen source, the presence of L-leucine greatly increases the growth rate even at concentrations so low that its contribution as nitrogen donor is unlikely to be important.


ICMSF methods studies. VII. Indicator tests as substitutes for direct testing of dried foods and feeds for Salmonella.

The correlation between the occurrence of indicator organisms and salmonellae in the same sample homogenate of various classes of dried foods was studied. In the first phase of the work involving 5837 samples, 1943 contained fecal coliforms and 100 contained Salmonella. Fecal coliforms were not detected in 53 of the Salmonella-positive samples. In the second phase, an additional 637 samples were tested for the presence of Enterobacteriaceae, coliforms, fecal coliforms, and Salmonella. Enterobacteriaceae were detected in 538 samples, coliforms in 506, fecal coliforms in 343, and Salmonella in 49. All Salmonella-politive samples were also positive for Enterobacteriaceae and coliforms. However, 6 of 49 Salmonella-positive samples were negative for fecal coliforms. Therefore, the use of the fecal coliform test as an index of Salmonella contamination would result in the rejection of Salmonella-negative foods and the acceptance of Salmonella-positive foods. It is concluded that none of these indicator groups are reliable as an index of Salmonella contamination in dried foods.


Isolation of bacteriophage lambda containing yeast ribosomal RNA genes: screening by in situ RNA hybridization to plaques.

We have developed an in situ hybridization technique which can be used to screen large numbers of hybrid bacteriophage for the presence of a particular inserted DNA sequence. Plaques of hybrid phage are formed on E. coli lawns on nitrocellulose filters, and their DNA is released, denatured, and fixed directly on the filters for hybridization to radioactive RNA probes. We have used this technique to isolate a number of hybrid bacteriophage lambda which contain EcoRl restriction fragments of the ribosomal RNA genes from yeast, and have examined the DNA from several of these phage.


Association of the folded chromosome with the cell envelope of E. coli: characterization of the proteins at the DNA-membrane attachment site.

Gentle lysis of E. coli cells in the presence of a DNA counterion (either 1.0 M NaCl or 5 mM spermidine) permits the isolation of the folded intact bacterial chromosome associated with membrane fragments. Most of the proteins in these chromosomes are also found in purified membrane preparations, and they can be identified as belonging to either the inner or the outer bacterial membrane. Ultraviolet irradiation of the membrane-attached chromosomes causes the formation of a stable complex between two inner membrane proteins (molecular weight 80,000 and 56,000 daltons) and 5-bromodeoxyuridine (BrdU)-substituted DNA. The photochemical attachment of BrdU-substituted DNA to specific membrane proteins suggests that these proteins may be bound to the DNA in vivo. Such DNA-membrane-binding proteins may have a role in the attachment of the folded chromosome to the bacterial envelope.


Mutagenic activity of carcinogenic and noncarcinogenic nitrofurans and of urine of rats fed these compounds.

The nitrofurans, 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2), N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT), nitrofurantoin, 5-nitro-2-furoic acid, 5-nitro-2-furamidoxime, 5-nitrofurfurylidene diacetate and the urine of rats fed these compounds, were assayed for mutagenic activity in Salmonella typhimurium strains TA100 and TA100FR1. All the nitrofurans were mutagenic in the order: AF-2 and FANFT greater than nitrofurantoin greater than 5-nitro-2-furamidoxime greater than 5-nitrofurfurylidene diacetate greater than 5-nitro-2-furoic acid. Strain TA100 was more sensitive than TA100FR1 to the mutagenic influence of these nitrofurans. Only the urine of rats fed AF-2, FANFT and nitrofurantoin had mutagenic activity. Again, TA100 was more sensitive than TA100FR1. The mutagenicity of the urine was not increased by treatment with beta-glucuronidase. AF-2, 2-amino-4-(5-nitro-2-furyl)thiazole (deformylated product of FANFT) and nitrofurantoin were excreted in the urine of rats fed these compounds; whereas the other nitrofurans were not excreted.


[Extramucosal single-layer suture with multifilament stainless steel wires in surgery of the colon. Experimental research].

With a view to evaluating whether and how far a fuller case documentation may contribute to a better knowledge of the real possibilities of multifilament stainless steel wires in surgery of the colon, the Authors present and discuss the results they obtained during the course of experimental research on extramucosa single-layer sutures. In the light of their personal histological documents, they emphasize the absolute biological inertia of this suture material, a property which, together with the capacity of not suffering the colliquative action of germs, means that multifilament stainless steel wires can be given full rights of domicile in colon surgery.


Single-passage extraction and permeability estimation of sodium in normal dogs lungs.

Bolus injections of 24NaCl, 131I-albumin, and indocyanine green dye were made into the right atria of anesthetized, ventilated dogs. Blood was sampled from the femoral artery, and from the dilution curves, instantaneous extractions, E2(t), and area averaged extractions, E3(t), were calculated for sodium at various plasma flows (Fp). Flow reduction was produced by transient inferior vena cava obstruction. E2(t) and E3(t) within any dilution curve generally started off with a high value, then decreased with time. The E3 that occurred at the peak of the albumin-dilution curve were about 0.11 for plasma flow of 0.75 liter/min and tended to decrease as flow increased. Parallel study of the sodium extravascular space at equilibrium gave values of 0.3-0.4 g of plasma sodium per g of tissue, suggesting that this volume is not infinitely small. Since the E was low it is unlikely that the high initial E(t) and the decreasing E(t) were due to early back flux. Calculation of capillary permeability surface area product [PS = Fp loge (1 - E)] showed an increasing PS with plasma flows. The injection of 9-mum microspheres at low and high total blood flow gave evidence supporting a decrease of capillary surface area (S) with decreasing total blood flow. Regional pulmonary blood flow also showed marked heterogeneity. Because of the low average extraction of sodium in the lung, insensitivity of the method in normal lungs cannot be excluded.


Vasomotor control of capillary transit time heterogeneity in the canine coronary circulation.

A set of indicator-dilution studies of the coronary circulation of the intact functioning heart was carried out in the dog to provide a data base for defining the effect of changes in vasomotor control on the exchange of materials across the myocardial capillaries. The reference substance used was 125I-labeled albumin; the diffusible substance used was 14C-labeled sucrose. Model analyses of the data were carried out. In previous models of the capillary exchange in the coronary circulation, it had been assumed that a single capillary transit time is representative of the whole. The data acquired here indicate that there is a very large heterogeneity of capillary transit times in the intact heart, and that the single transit time model is approximately true only when the resistance vessels are maximally dilated. The present pattern of findings is explained best by a model of the coronary microcirculation based on capillary-large vessel units with a variable heterogeneity of flow or capillary lengths, hence of transit times. We conclude that a major determinant of the extraction of each diffusible substance, on a microscopic level, is the distribution of capillary transit times.


Effect of steric restriction on the intracortical distribution of microspheres in the dog kidney.

The effect on the intracortical distribution of microspheres and radioactivity caused by steric hindrance of the free movement of spheres into afferent arterioles are described by two mathematical models. The results are compared with corresponding experimental data obtained in six kidneys from normotensive dogs. The first model (A) assumes that spheres are distributed as blood flow, regardless of their size, except for those having diameters greater than that of an afferent arteriole and which do not enter this vessel. The second model (B) includes the Ferry correction. The experimental data show that the percent recovery of spheres with diameters of 20-25 mum was significantly greater in the outer cortex and significantly less in the juxtamedullary cortex than recovery of the smaller spheres, and that the distribution of spheres with diameters of 10 mum to about 17 mum seems uninfluenced by the sphere size. The experimental results we have obtained fit best with model A. We found that according to both models steric restriction is a factor of major importance in relation to the intracortical distribution of spheres, and the analysis shows that the blood flow in the inner part of the renal cortex is grossly underestimated by the method of isotope labeled microspheres when diameters of 15 +/- 5 mum are used in the dog. Furthermore we found that dilation of the afferent arterioles will change the steric hindrance so that a redistribution of spheres and radioactivity may occur without any redistribution of blood flow. It is suggested that the results interpreted as redistribution of blood flow can be explained as due to altered steric hindrance alone, i.e., as a methodological artifact.


Echocardiographic patterns of ventricular contraction in the Wolff-Parkinson-White Syndrome.

Echocardiograms of 52 patients with the Wolff-Parkinson-White (WPW) syndrome were investigated. Abnormal patterns of motion were observed in the left ventricular posterior wall (LVPW) and the interventricular septum (IVS). The abnormal LVPW motion was presumably specific for the syndrome and included early onset of the anterior motion which preceded the first heart sound (S1) and a premature peak formation of the anterior displacement which occurred before the second heart sound. The latter was usually followed by a second lower peak. These findings seem to suggest that both contraction and relaxation of LVPW begin earlier than normal due to ventricular pre-excitation initiated at or near the LVPW. The IVS abnormality characteristic of the syndrome was a posterior protrusion, the onset of which preceded S1. Subsequent motion of IVS was either paradoxical or hypokinetic during ejection. These IVS abnormalities were quite similar to those observed in left bundle branch block. The abnormal LVPW motion was observed in all 20 Type A patients and 10 of the 32 Type B patients. The abnormal IVS motion was observed in 10 Type B patients. We feel that echocardiography could be used as an auxiliary noninvasive means of locating the pre-excitation site in patients with WPW.


Intracardiac electrography in endocardial cushion defects.

Conduction of the sino-atrial impulse from the high right atrium to the ventricles was studied by intracardiac electrography in 21 unoperated patients, age 3 months to 11 years, with endocardial cushion defects (ECD). The high right atrium-to-low right atrium conduction time was prolonged in 15 of 18 subjects (mean 57 +/- 20 msec). The low right atrium-to-His bundle conduction time (LRA-H) was normal in 16 of 17 subjects (mean 82 +/- 30 msec), prolonged in one. The His-to-ventricle conduction time (H-V) was normal in 16 of 17 subjects (mean 37 +/- 8 msec), equivocally short in one. Nine patients with ECD, age 3 to 21 years, were studied postoperatively. One had an acquired complete atrioventricular block in the His bundle. Two had prolonged LRA-H and two prolonged LRA-H and two prolonged H-V. The surface ECG failed to identify accurately either prolonged atrioventricular conduction or the site of prolongation.


Noninvasive assessment of mitral insufficiency by transcutaneous Doppler ultrasound.

Instantaneous aortic arch blood velocity was recorded transcutaneously from the suprasternal notch, using a 2.2 MHz Doppler ultrasound unit, in 18 normals and 16 patients undergoing cardiac catheterization who had murmurs of mitral regurgitation. In normals aortic blood velocity rose rapidly in early systole to a midsystolic peak then fell to zero velocity. These roughly parabolic patterns had area ratios beneath the first and second halves of the curves measuring 52:48 +/- 3 (SD). With increasingly severe mitral regurgitation the pattern became skewed leftward such that the percent in the first half of systole ranged from 53-79%. From the angiograms of our sixteen patients an estimate of true percent regurgitation was made using the Fick cardiac output and ventricular volume measurements. When compared with the area under the first half of the velocity curve a strong correlation was found (r = 0.84) indicating that this Doppler technique can be used to evaluate mitral insufficiency.


Acute hemodynamic interventions shift the diastolic pressure-volume curve in man.

Frame-by-frame analysis of angiograms in 16 patients revealed that hemodynamic interventions are capable of producing substantial shifts in the diastolic pressure-volume curve. Angiotensin raises blood pressure and shifts the entire pressure-volume curve up, and nitroprusside lowers blood pressure and shifts the curve down. Indirect measurements of pleural pressure in seven patients (via esophageal pressure) showed that pleural pressure changes were too small to account for these shifts. Analyzing our results in terms of a theoretical pressure-volume equation previously validated in dog studies did not show the observed shifts to be the product of acute changes in the elasticity of the myocardium itself. This same analysis suggested that indirect changes in the external mechanical constraints acting on the left ventricle such as the right ventricular pressure, the pericardium, and perhaps viscoelastic effects related to changes in filling rate account for the pressure-volume curve shifts with intervention. The fact that one cannot in general relate a specific volume to a given pressure in the face of hemodynamic interventions calls into question the use of end-diastolic pressure interchangeably with end-diastolic fiber length when interpreting systolic events in terms of the Frank-Starling mechanism.


Response to exercise in patients after total surgical correction of Tetralogy of Fallot.

Heart rate, blood pressure, physical working capacity, and electrocardiographic changes were evaluated during upright bicycle exercise in 43 asymptomatic patients, aged seven to 41 years, one to 14 years after total surgical correction of tetralogy of Fallot (TF). One hundred and nine normal subjects between the ages of five and 42 years served as controls. The patient and control groups, subdivided by sex and body surface area (BSA), were similar in height and weight. When comparing males to males and females to females with BSA greater than or equal to 1.2 m2, maximal heart rates and working capacities were lower in the patient groups than in the control groups. An inverse relationship was observed between maximal working capacity and age at surgery in both male and female patient groups. By contrast, especially in the males with BSA less than 1.2 m2, the mean maximal heart rates and working capacities did not differ significantly between the patient and control groups. Premature atrial or ventricular contractions were recorded in ten of 43 patients (23%) after exercise. Five of these ten patients had multifocal premature ventricular contractions (PVC) and four had unifocal PVC. In the five patients with multifocal PVC, a short burst of ventricular tachycardia occurred in two, coupling in one, and bigeminal rhythm in two. Cardiac arrhythmia was not observed in the control group. Although our current surgical results are excellent, this study suggests that impaired cardiovascular function persists after corrective surgery and that early surgical treatment may be more desirable. Furthermore, additional data suggest that the exercise procedure may be useful in detecting and managing patients who may develop life-threatening arrhythmias following intraventricular surgery.


Effects of visibility of a prepalatal cleft  on the evaluation of speech.

Sixty speech pathology students participated in a double-blind factorial experiment to assess the impact of a distorted lip and nares on the evaluation of various speech dimensions. The subjects were randomly assigned to one of four treatment groups. All Ss read identical case studies about a child born with a cleft. One of two photographs was attached to each case study. One was an accurate photography of a girl with post-operative disfigurement. The second was of the same girl, but it had been retouched to eliminate the disfigurement. Half of the Ss in each photograph condition listened to a recording of a normal speaker while the other half of the Ss listened to a nasal speaker. The results of various analyses indicated that a visible disfigurement significantly affected the Ss evaluation of nasal speech, increasing their perception of nasality and of the need for speech therapy.


Treatment of cleft lip and palate during the Revolutionary war: bicentennial reflections.

Cleft palate surgery was not performed either in Europe or in America during the time of America's Revolutionary War. Cleft lip surgery was performed by the pinning and figure-of-eight thread method of closure in keeping with current European teachings on the subjects. This surgery was frequently performed by itinerant mendicants, charlatans, and also by the more legitimate members of the surgical community living in the 13 states at the time of the Revolution. A brief review of the surgical methods employed form the major portion of the article.


Congenital bilateral anorchia in childhood: a clinical, endocrine and therapeutic evaluation of twenty-one cases.

An evaluation of twenty-one boys, including a discordant pair of identical twins, is presented in whom bilateral anorchia was found with a negative family history and without history of breech presentation or of postnatal testicular trauma, torsion or orchitis. The most likely cause is prenatal testicular torsion. The incidence of the condition in our hospital is 1 in 177 cases of cryptorchidism. Prepubertal growth was normal before treatment, and testosterone replacement therapy allowed a normal pubertal growth spurt and skeletal maturation. Although demonstrable basal urinary testosterone was found in the subjects with a postpubertal bone age, most patients tested showed no increase after stimulation with human chorionic gonadotrophin. In the presence of a normal penis and scotum, such findings, together with a high basal FSH and an increased response of plasma LH to LHRH, make surgical exploration unnecessary. In the rare patient who shows a positive but subnormal response of testosterone to HCG, Leydig cells are presumed to be present either ectopically or in rudimentary testes, and further surgical exploration is indicated.


Effect of oestradiol on gonadotrophin release induced by LHRH in men.

In order to study the role of the oestradiol on the feed-back mechanism, fourteen men were subjected to LHRH test before and after treatment with oestradiol. The oestrogen was administered i.m., 3 mug/kg/24 h and 150 mug/kg/24 h, in two groups of patients. Doses of 3 and 150 mug respectively produced a reduction of LH release of 74% and 60% below control. FSH response to LHRH was almost abolished by oestrogens. Our findings show that oestrogens in men decrease the pituitary response to LHRH.


Significance of genetic factors for the plasma insulin response to glucose in healthy subjects.

The intravenous glucose tolerance and plasma insulin response to glucose infusion were analysed in a twin and family material, comprising 279 healthy subjects. The relation between the blood glucose and plasma insulin values was studied by an analysis of the principal eigenvalues. The variables obtained were corrected for sex, age and weight, and standardized with regard to mean and variance. The results showed that at least four of the variables have appreciable familial correlations, corresponding to a heritability (h2) varying between 0.38 and 0.72. These correlations could not be accounted for by common environment alone. Thus, the beta cell function in normal man, as measured by a glucose challenge test, appears to be genetically regulated.


Congenital absence of gluteal muscles. Report of two sibs.

A family is described with the following features: 1) Two propositi, a male and a female, with congenital absence of gluteal muscles and with spina bifida occulta. 2) Both parents and two apparently normal siblings with sacral spina bifida occulta. 3) Two siblings of the propositi who died soon after birth, one with anencephaly and the other with a probable spina bifida. Two alternative hypotheses for the etiology of these malformations are suggested: first, the muscular defect could be caused by an autosomal recessive gene independent of the open neural-tube defects; second, both types of malformations could be due to the same autosomal recessive gene. Then compensatory muscular changes which allow the propositi to walk are discussed.


Autosomal reciprocal translocations and 13/14 translocations: a population study.

Fifteen children with autosomal reciprocal translocations were found in an incidence study of chromosome aberrations among 11,148 consecutively liveborn children in a Danish Maternity Hospital (1.34 per 1,000). The segregation rate of the balanced forms was 60% and that of unbalanced forms 4%; the frequency of familial cases was 73%. None of the probands or their relatives who carried the balanced translocation had any physical or mental abnormalities which could be associated with the chromosome aberration. We found two children with unbalanced translocations among the 45 relatives examined, but more studies of unselected population groups are needed before any segregation rates for unbalanced derivatives of autosomal reciprocal translocations can be estimated. We found a significantly higher frequency of abortions, stillbirths and perinatal deaths in the progeny of carriers with the translocation, compared with non-carriers in the same families. The unbalanced forms of autosomal reciprocal translocations in most cases are probably not compatible with the development of a foetus to a liveborn, healthy child. Fifteen children with 13/14 translocation were found among 11,148 consecutively liveborn children. Incidence, segregation rates and type of translocation are discussed. Fertility and risk for carriers of D/D translocations of producing children with autosomal trisomy are discussed.


A membrane-associated autoantibody in a case of myasthenia gravis with chronic hepatitis.

A new antibody reacting with a cell membrane-related antigen is described in a patient with myasthenia gravis and chronic liver disease. The antibody gave a striking peripheral immunofluorescence in all cells tested and did not react with smooth muscle fibres, being distinct from anti-actin and anti-myosin antibodies. The antigen was widely distributed in human and animal tissues. The exact relationship of this antibody to the clinical condition is not obvious but its presence is further evidence of widespread disturbances of humoral immunity in myasthenia gravis.


Liver cell surface localization of hepatitis B antigen and of immunoglobulins in acute and chronic hepatitis and in liver cirrhosis.

This paper describes immunofluorescence studies on liver cell surface localization of hepatitis B surface antigen (HBsAg) and of IgG in acute and chronic hepatitis and in cirrhosis. In acute hepatitis B, HBsAg was found at the surface of hepatocytes in an early phase of the disease, but not during the recovery. This finding is consistent with the hypothesis that immune reactions to HBsAg may be responsible for the liver cell lysis. In HBsAg-positive chronic hepatitis and cirrhosis the antigen was found in the cytoplasm, but not on the surface of the hepatocytes, while in HBsAg-negative cases the antigen could not be detected in the liver cells. Both in HBsAg-positive and in HBsAg-negative chronic active hepatitis (CAH) and cryptogenic cirrhosis IgG bound to the membrane of the hepatocytes could be detected, suggesting a role of antibodies in the pathogenesis of the disease.


Studies on the pathogenesis of experimental autoimmune renal tubulointerstitial disease in guinea-pigs. III. The role of adjuvants in the induction of disease.

Experimental renal tubulointerstitial disease is induced in guinea-pigs by anti-tubular basement membrane autoantibodies. The complete mechanism of mononuclear cell accumulation in the target organ is not known; however, in addition to antibody, complement and radiosensitive leucocytes are important. In the present experiments we explored the influence of adjuvant in the accumulation of mononuclear cells in kidneys of actively or passively immunized guinea-pigs. We found that renal disease could be induced without adjuvant, by multiple injections of rabbit tubular basement membrane. Lesions were comparable to those groups which received a single dose of antigen in Freund's complete or pertussis vaccine adjuvant. Passive transfer of nephritis confirmed that adjuvant is not necessary for either the accumulation of mononuclear cells or the formation of antibodies with particularly potent pathogenicity.


Heterogeneity of stem cells in severe combined immunodeficiency.

Two patients with severe combined immunodeficiency disease (SCID) having variable B-cell development have been shown to have marrow precursors of lymphoid cells which can be induced in vitro by thymic factors to express certain T-cell surface characteristics (HTLA+ phenotypes). Their marrow cells could not, however, be induced by these same factors to develop the E-rosette marker or functional activities of T lymphocytes. The marrow of these children also showed, when compared to that of normal adults, a different distribution of cellular elements on density gradient fractionation. The findings support the view that the disorder under study has a different pathogenesis from other forms of SCID previously analysed.


The Crithidia luciliae kinetoplast immunofluorescence test in systemic lupus erythematosus.

An immunofluorescence test for antibodies to native DNA, using the kinetoplast of Crithidia luciliae as substrate, has been assessed in comparison with the Farr precipitation technique, on a total of 395 sera from sixty-three patients with systemic lupus erythematosus, 185 other hospital patients and sixty healthy controls. The immunofluorescence test appears to have great specificity as a diagnostic test for SLE, though lacking the sensitivity of the Farr technique. Like the latter, it is altered by immunosuppressive treatment, and in patients with SLE nephritis on immunosuppression it does not show good correlation with activity of renal disease. Its specificity and simplicity nevertheless make it a valuable clinical test.


Chronic mucocutaneous candidiasis accompanied by enhanced antibody production.

In four cases of chronic mucocutaneous candidiasis (CMCC) enhanced antibody production against candida antigen was detected in association iwth depressed cellular immunity. These patients showed extremely high agglutinin titre to candida antigen, while they were unable to mount delayed hypersensitivity reactions to candida antigen, tuberculin-purified protein derivative (PPD) or to dinitrochlorobenzene (DNCB). They also showed prolonged survival of skin homografts. There was no correlation between migration inhibitory factor (MIF) production and blast transformation in response to candida antigen.


Cutaneous melanoma of the head and neck.

Melanoma of the head and neck, if diagnosed early and treated with aggressive appropriate surgical therapy, is potentially curable in up to 80 to 90 per cent of the cases. The use of microscopic staging by level of invasion and thickness of the tumor is helpful in determining the appropriate surgical procedure for the individual patient and is of prognostic significance. If possible, prophylactic incontinuity regional node dissection should be performed for melanomas of the head and neck that have invaded to Level III or deeper, especially those that are greater than 1.5 mm in thickness. The histological status of the regional nodes is beneficial both therapeutically and prognostically, in that patients who have negative nodes have a better prognosis than those with microscopically positive nodes. Also, these patients with microscopically positive nodes have a much better survival than those with macroscopically positive nodes. Melanoma of the head and neck should be treated very aggressively with wide excision of the primary tumor in order to prevent local recurrence and further spread of the disease. Since surgical treatment is the only effective curative measure for melanoma, all localized tumor in the region of the primary and solitary distant metastasis should be removed if possible. The adjunctive use of chemotherapy and immunotherapy when regional nodes are involved with melanoma is being studied and may be of some benefit. The combinations of surgical therapy, chemotherapy, immunotherapy, and radiotherapy offers the patient with advanced disease significant palliation, sometimes for prolonged periods. The treatment of head and neck melanoma is best summarized by the statement in the December 4, 1965 of The Lancet.-.29 "The surgeon who first operates on a malignant melanoma has a great responsibility. Prompt and competent action will give the patient a chance of survival better than in most other forms of cancer. The only additional operative surgical skill required is the ability to cut and apply split skin grafts. If he lacks confidence therein, let the surgeon refer the case at once and certainly before he has ruined, by niggling interference, the patient's chance of survival."


Microsurgical composite tissue transplantation: a method of immediate reconstruction of the head and neck.

A new approach utilizing microsurgical composite tissue transplantation is advocated for the management of difficult reconstructive problems in the region of the head and neck. This method is especially useful when local flap coverage may be inadequate. Morbidity and hospitalization is reduced and reconstruction is complete upon discharge from the hospital. Extirpative surgery for a malignancy can now have the defect reconstructed immediately with less disfigurement and psychological insult to the patient and family.


Cancer of the nose: ablation and repair.

Cancer of the nose is very common and lesions seen by reconstructive surgeons are often recurrent and extensive. Surgical removal of cancer of the nasal skin can usually be accomplished under local anesthesia, and in most instances frozen section histologic examination should be used to confirm the adequacy of excision. The location and three dimensional extent of the tumor will dictate the choices of repair or reconstruction as well as the timing thereof. Very small lesions can be excised with primary closure; other well circumscribed tumors can be excised and the defect closed with an appropriate nasal flap. In our experience most nasal skin cancers have been managed by excision and full-thickness skin grafting. We have found the skin of the neck and that of the preauricular region to provide the best skin cover except in the upper third of the nose where upper eyelid skin provides excellent coverage. We have used composite grafts from the ear to replace up to two-thirds of an alar rim. Nasolabial, cheek, and midline forehead flaps are useful in a variety of instances, but usually when less than one-half of the nose has been excised. We have been pleased, in most instances, with the Converse scalping flap for near total, subtotal, and extensive three-dimensional lower nasal defects. (Transverse superficial temporal artery pedicle flaps can be successfully used to reconstruct large nasal defects with adjacent cheek loss.) We have rarely used distant flaps. Regardless of what regional pedicle flap has been transferred to the nose, subsequent revisions of a relatively minor nature will nearly always enhance the result. Patients who have undergone extended total nasectomies are probably best managed with a prosthesis, as prognosis is often guarded and flap reconstruction may be quite unsatisfactory. In our experience, defects in lining and support can usually be repaired with local nasal tissue.


Facial resurfacing with the Limberg flap.

The Limberg flap is a valuable tool in teaching the art of local flap design. It is a very versatile addition to the armamentarium of any surgeon which provides a precise mathematical design for closure of both the primary and the secondary (donor) defect. The lines of maximum extensibility of the skin are crucial in the design of the rhomboid and the flap. Attention to these details avoids complications and assures good aesthetic and functional results.


Bone blood flow in the rat determined by the uptake of radioactive particles.

1. Two different suspensions of radioactive particles, 99Tcm-labelled microspheres and 59Fe-labelled resin particles, were injected into the rat central circulation in order to study the distribution of blood flow to the skeleton. 2. Measurements of particle uptake showed that the mean proportion of cardiac output flowing to the skeleton was 3-1%, a figure much lower than the valve of 27% found by Brookes (1970) in a similar experiment. 3. The blood flow per unit mass to individual bones covered a wide range.


Dental visits, teeth remaining, and prosthetic appliances in a Norwegian urban population.

Of a random sample (177 persons) of 35-year-old Oslo citizens, 117 subjects (66.1%) attended a dental examination. Questionnaires were returned by mail from 28 of the non-respondents. The most common reason for not attending was "difficulty in finding time for an appointment." About 89% of the respondents had visited a dentist regularly. Regular dental care seemed to be more common among people with a high level of education than among those with a low level of education. The most common treatment performed at the last dental visit was filling of cavities. The mean number of remaining teeth was 25.7. Females in the lower educational group had significantly less teeth than those in the higher educational group. No subjects were totally edentulous, but five persons had one edentulous jaw. Nine full or partial dentures were found. Twelve percent of the subjects had one or more fixed bridges, and 29.9% of the persons had one or more crown restorations. There seemed to be no differences between the sexes or educational levels in this respect.


Efficiency in clinical trials of caries preventive agents and methods.

The idea sequence of tests required to establish a caries prophylactic agent or procedure on a public health basis is described and a clear distinction is drawn between an experimental clinical trial, carried out under ideal conditions, and a community clinical trial carried out under real life conditions. Because of the artificial nature of an experimental clinical trial there are strict limitations placed on its objectives and the inferences that can be drawn from its results. Bearing these limitations in mind and using the data collected during a 3-year trial of a fluoride mouthrinse, two methods of improving the efficiency of experimental clinical trials of caries prophylactic agents are considered. Firstly, a reduction in length is suggested either by planning the trial to run for a shorter period in the first place or, alternatively, by stopping the trial when the criteria for success have been satisfied. Secondly, it is proposed that the number of subjects participating in the trial be reduced by excluding those whose contributions to the aims of the trial are likely to be low.


Franz Kafka.

The hereditary and environmental influences on the genius of Kafka's personality are discussed. His exterior, somatic constitution is represented by photos taken during his lifetime as well as some posthumous, imaginary portraits and sculptures by various, mostly Prague, artists. The concept of the artist's psychic formation - his mental makeup - is documented from his diary, dreams, letters, literary work and reports by his friends. The whole proves his introversion, ambivalence, hypersensitivity, obstinancy, anxieties, behavioral anomalies, a life rich in fantasies and his underestimation of his own literary work. (He speaks about it as an idiosyncrasy, wishing it to be burned after his death.) Some little-known drawings by Kafka complement the picture of his talents and creativity. The psychological and psychopathological roots of his extraordinary position in the cultural awareness of his epoch are discussed.


Transformations of consciousness. A cartography, II. The perception-meditation continuum.

The Japanese Zen and Indian Yoga meditation techniques of zazen, dhārnā, dhyān, savichār- and nirvichār-samādhi are cartographed on a perception-meditation continuum of increasing trophotropic arousal. Certain psychophysiological characteristics and experiential dimensions of these hypometabolic states are discussed and it is proposed that observer-observed interactions or trans-substantiation (commonly termed 'reality') may not be subject to Goedelian restrictions if they proceed in hyper- and hypoaroused states. The meaning of aroused, non-verbal and logically non-Aristotelean states cannot be conveyed to individuals in a non-aroused state. The converse is also true when the relationship of arousal levels is reversed.


Pictorial expression of ulcerative colitis.

The most prevalent emotional characteristics of patients suffering from ulcerative colitis are illustrated in pictures spontaneously drawn by a young patient while hospitalized. With the onset of the illness, the pathological content of the drawings surfaced. Using anthropomorphism of animals, the patient was able to identify with the story-book characters and reveal the problems of her own inner world and masked depression, which she was not able to express otherwise. Although she viewed herself at the center of the world, innocent and sensitive, she felt inferior and estranged from society. In defense, she withdrew from the hostile world and tried to hide self-directed aggression.


Comparative study on gastric secretory function between benign gastric polyp and elevated type of early stomach cancer with special reference to histological findings of gastric mucosa.

In cases of elevated type of early cancer and benign polyp of the stomach, gastric analysis was performed and the results were compared with histological findings of gastric mucosa. Both clinical entities were found to show lowered acid and pepsin secretory functions reflecting severe and extensive atrophic changes of gastric mucosa. Dissociation in secretory disturbance between acid and pepsin was also noticed.


Functional preservation of the mammalian kidney. IV. Functional effects of perfusion with dimethyl sulfoxide (DMSO) at normothermia.

Rabbit kidney were perfused at 37degreesC with various concentrations of DMSO in a K+-Mg2+-rich perfusate. The effects of DMSO on various functional parameters of the rabbit kidney perfused for 60 min were compared with the functional effects of perfusion without DMSO under the same conditions. DMSO produced deviations in vascular resistance and perfusate flow rate from control values. In kidneys perfused with 1.4 and 2.8 M DMSO these vascular changes resulted in changes in GFR at relatively unchanged filtration fractions. The closely parallel relationship between changes in GFR and urine flow in all groups indicates that perfusion per se or perfusion with DMSO may shift the regulation of urine flow rate from tubular reabsorption, which obtains in the in vivo situation, to glomerular filtration. This view was supported by the relatively unchanged parameters of Na+ reabsorption and fractional water excretion during perfusion with all concentrations of DMSO. Additionally, DMSO perfusion resulted in significantly greater weight gains than those observed in kidneys perfused without DMSO, and significantly depressed clearances of PAH, with 2.1 and 2.8 M DMSO.


The effect of freeze rate, duration of phase transition, and warming rate on survival of frozen canine kidneys.

The effect of cooling rate, warming rate, and duration of phase transition upon survival of frozen canine kidneys was investigated. In the present study, 11 kidneys out of 14 rapidly cooled (2--4degreesC/min) to --22degreesC and thawed (70--110degreesC/min) were viable following contralateral nephrectomy. The serum creatinine and BUN levels rose to a maximum of 8.4 and 30 mg%, respectively, on the eighth day post-contralateral nephrectomy. Average survival time was 10 days; however, two of the dogs in this group were allowed to survive, one for 3 months and one for over 2 years. Eight kidneys out of 16 slowly cooled (0.25-1.0degreesC/min) and either rapidly or slowly warmed (20-30degreesC/min) had function to produce small amounts of urine; however, they did not survive more than 5 days after contralateral nephrectomy. Cooling rates of 0.1 and 10degreesC/min were too harmful to the kidney to have renal function after reimplantation. The minimum renal cell damage as assessed by LDH and GOT in the post-freeze perfusate was found in the 2-4degreesC/min cooling rate following rapid warming (70degrees-110degreesC/min). Correlation of the duration of phase transition time to renal cell damage was linear for LDH and GOT (r=0.93). This result suggests that the duration of phase transition time also is an important factor during the freezing process, affecting post-thaw survival of canine kidneys.


Pulmonary function after recovery from the adult respiratory distress syndrome.

Eight adult patients with acute respiratory distress syndrome were treated with positive end-expiratory pressure (PEEP) ventilation. The results of detailed examinations of pulmonary function in these eight patients obtained after different periods of time following discharge from the respiratory care unit were analyzed to determine the degree of disturbances in pulmonary function. For comparison, examinations of pulmonary function were also performed on two patients who were treated with zero end-expiratory pressure ventilation. This follow-up study showed remarkably few abnormalities. Whe present, restrictive disturbances in pulmonary function, especially decreased static compliance and diffusing capacity, were found. No correlation was found between the pulmonary-function results of the eight patients and the duration of the PEEP treatment, nor between the results and the time interval between treatment at the respiratory care unit and the moment of the pulmonary function studies.


Treatment of bronchopleural fistula during continuous positive pressure ventilation.

Pneumothorax as a complication of continuous positive pressure ventilation may result in the formation of bronchopleural fistula. If positive end-expiratory pressure cannot be sustained, the functional residual capacity and the ratio of alveolar ventilation to perfusion may decrease, and pulmonary gas exchange may be severely impaired. Recommended therapy includes removal of positive airway pressure and institution of negative intrapleural pressure via thoracostomy tube, but this may cause severe hypoxemia in patients with acute respiratory failure in spite of potentially toxic inspired oxygen concentrations. We applied positive intrapleural pressure equal to the end-expiratory airway pressure of three patients who developed bronchopleural fistula during therapy for acute respiratory failure. Positive intrapleural pressure facilitated resolution of the bronchopleural fistula in each case.


The tethered spinal cord: its protean manifestations, diagnosis and surgical correction.

31 children with spina bifida occulta who have presented with back pain, scoliosis, a progressive neurological deficit involving lower limbs or a neurogenic bladder have been found to have a tethered spinal cord. Release of the tethered spinal cord has always relieved pain, frequently occrected a progressive scoliosis, and arrested or improved neurogenic foot deformities as well as neurogenic bladder.


[Lymphocele following renal transplantation].

Lymphoceles as seen after renal transplantation are cystic swellings filled with lymph, situated beneath the lower end of the transplant on the iliac vessels. It seems that the lymph originates from lymphatics cut during operation. 5 such cases have been seen in our series of 268 renal allotransplantations. Symptoms arose 1 to 6 months after transplantation. According to our own experience and to 39 cases of the literature, deterioration of renal function because of compression of the ureter is the most common symptom. Therefore, intravenous pyelography is the main diagnostic means. Some cysts can be visualized by lymphangiography. Marsupialisation to the peritoneal cavity is the treatment of choice.