Plasma concentrations of diazepam and desmethyldiazepam during chronic diazepam therapy.

1 Plasma concentrations of diazepam and its metabolite, desmethyldiazepam, have been measured in both in- and out-patient groups treated with diazepam for periods varying between 1 month and 10 years. 2 The diazepam concentration was directly related to the dose of diazepam ingested and inversely related to the age of the patient. 3 A highly significant relationship was obtained between the concentration of desmethyldiazepam and diazepam. 4 The plasma concentrations of both diazepam and its metabolite were independent of sex, duration fo therapy and patient group.


Sterelogical analyses of leukaemic cells.

With the development of different treatment schedules for acute and chronic leukaemias it has become essential that a uniform, standardized classification of malignant cell type is achieved. Without a standardized diagnosis it is impossible to compare the results of differing treatment regimes. Present diagnosis is still based on clinical criteria and subjective assessment of cell morphology; objective and quantitative data are required. Stereological analyses are capable of providing quantitative data. Stereological techniques have been applied to the study of malignant cell types in different leukaemias for the first time and quantitative data has been obtained. The nuclear and mitochondrial volume fractions of the cells have been estimated for normal lymphocytes, the lymphocytes of chronic lymphocytic leukaemia, acute lymphoblastic leukaemia, the abnormal cells of hairy cell leukaemia and several other haematological malignancies. The results clearly indicate that the cell types fo different leukaemias possess measurably different properties and that stereological methods can be used to investigate the leukaemic cell quantitatively.


Measles retinopathy during immunosuppression.

A 14-year-old boy had an acute attack of measles while on cytotoxic chemotherapy for a testicular neoplasm. Two months later a fatal measles encephalopathy developed, verified by histological, ultrastructural, and immunofluorescent studies. Ophthalmoscopy showed progression of flat depigmented areas of the retina as well as prominent lesions mimicking central serous retinopathy or retinitis. Histopathological studies showed focal retinal necroses with invasion of pigment-laden macrophages into the retina. There were no inflammatory cell infiltrations in the choroid. The presence of structures with the morphological and antigenic properties of measles virus in the affected areas of the retina was shown by electron microsocpy and indirect immunofluorescence. With the increase of immunosuppressive therapy for various purposes the incidence of opportunistic measles virus infections is likely to rise. In the clinical management of such complications ophthalmological examination may prove very helpful.


Does enucleation of the eye containing a malignant melanoma prevent or accelerate the dissemination of tumour cells.

A reappraisal of survival data on patients with uveal melanomas has led us to these impressions: (a) that the mortality rate before enucleation is low, estimated at 1% per year; (b) that the mortality rate rises abruptly following enucleation, reaching a peak of about 8% during the second year after enucleation; and (c) that approximately two-thirds of the fatalities could be attributed to the dissemination of tumour emboli at the time of enucleation. From these impressions we believe the following conclusions are warranted: (a) enucleation as it has been performed in the past may have for many patients an adverse rather than a beneficial effect with respect to the development of metastatic disease from malignant melanoma of the choroid and ciliary body. (b) A long-term follow-up study of untreated patients with melanomas of the choroid and ciliary body is indicated. (c) New techniques for enucleation designed to prevent the dissemination of tumour cells must be developed and tested to enable the ophthalmic surgeon to remove safely the tumour-containing eye that has developed such complications as uncontrollable glaucoma, panophthalmitis, or proptosis from extraocular extension.


Selectivity of RNA chain initiation in vitro. 1. Analysis of RNA initiations by two-dimensional thin-layer chromatography of 5'-triphosphate-labeled oligonucleotides.

A method for the rapid and quantitative analysis of 5'-terminal oligonucleotides of RNAs made in vitro is described. The method involves synthesis of RNA in the presence of [gamma-32P]ATP or GTP, isolation of the RNA, and digestion with T1 or pancreatic ribonucleases to release labeled 5'-triphosphate termanated oligonucleotides. The oligonucleotides are then subjected to chromatography on a polyethyleniminecellulose thin-layer system using 2 M LiCl, 0.01 M EDTA (pH 6.5) in the first dimension and 1.5 M LiCl, 1.8 M formic acid, 0.005 M EDTA (pH 2.0) in the second. RNAs made with E. coli RNA polymerase and lambdacb2, T7, T4, and adenovirus 2 DNA yield characteristic fingerprint patterns. The utility of this method in studying selectivity of in vitro RNA chain initiation is discussed.


Selectivity of RNA chain initiation in vitro. 2. Correlation of 5'-triphosphate-labeled oligonucleotides on polyethyleniminecellulose thin-layer chromatography with RNA transcripts of bacteriophage lambda cb2 and T7.

Methods are described for the correlation of 5'-terminal oligonucleotides separated by two-dimensional polyethyleniminecellulose thin-layer chromatogrphy with specific RNA transcripts made in vitro from DNA of phages T7 and lambdacb2. The 5'-terminal oligonucleotides transcribed from DNA containing mutations and alterations which affect the RNA transcripts from specific promoters were compared with fingerprints of RNA from wild type DNA. Specific RNAs were purified on polyacrylamide gels and digested and their 5'-terminal oligonucleotides subjected to chromatography. Transcription of DNA fragments containing specific promoters was carried out and the 5'-oligonucleotide fingerprints of the RNA products compared with fingerprints of the RNA from the whole DNA. Using these methods and information known about T7 and lambdacb2 RNA 5'-terminal sequences, it was possible to identify many of the oligonucleotides separated on the polyethyleniminecellulose chromatography system.


Properties of 3-methyladenine-DNA glycosylase from Escherichia coli.

An Escherichia coli enzyme that releases 3-methyladenine and 3-ethyladenine in free form from alkylated DNA has been purified 2800-fold in 7% yield. The enzyme does not liberate several other alkylation products from DNA, including 7-methylguanine,O6-methylguanine, 7-methyladenine, N6-methyladenine, 7-ethylguanine, O6-ethylguanine, and the arylalkylated purine derivatives obtained by treatment of DNA with 7-bromomethyl-12-methylbenz[a]anthracene. The reaction of the enzyme with alkylated DNA leads to the introduction of apurinic sites but no chain breaks (less than one incision per ten apurinic sites), and there is no detectable nuclease activity with native DNA, depurinated DNA, ultraviolet-irradiated DNA, or X-irradiated DNA as potential substrates. The enzyme is termed 3-methyladenine-DNA glycosylase. It is a small protein, Mr = 19 000, that does not require divalent metal ions, phosphate, or other cofactors in order to cleave base-sugar bonds in alkylated DNA.


Studies of virus structure by laser-Raman spectroscopy. Turnip yellow mosaic virus and capsids.

Laser-Raman spectroscopy of the turnip yellow mosaic virus (TYMV) and its capsid indicate the following features of the structure and assembly of the virion. The secondary structure of coat-protein molecules in TYMV is comprised of 9 +/- 5% alpha-helix, 43 +/- 6% beta-sheet, and 48 +/- 6% irregular conformation and is not altered by the removal of the RNA from the capsid. Introduction of as many as 200 chain scissions per RNA molecule also does not affect the overall secondary structure of the encapsulated RNA, which is 77 +/- 5% in the A-helix form. Tryptophan and cysteine residues of the coat protein appear to be in contact with the solvent, while only one of three tyrosines per coat protein is available for hydrogen bonding of its p-hydroxyl group with H2O molecules. Both cytosine and adenine residues of TYMV RNA are protonated in substantial numbers near pH 4.5, suggesting elevation of their respective pKa values within the virion. The Raman data are consistent with chemical evidence favoring interaction between protonated bases of RNA and amino acid side chains of coat protein in TYMV.


Kinetic studies on the inactivation of Escherichia coli RTEM beta-lactamase by clavulanic acid.

The kinetic details of the irreversible inactivation of the Escherichia coli RTEM beta-lactamase by clavulanic acid have been elucidated. Clavulanate is destroyed by the enzyme and simultaneously inhibits it by producing two catalytically inactive forms. One of these is transiently stable and decomposes to free enzyme (k = 3.8 X 10(-3) S-1), while the other corresponds to an irreversibly inactivated form. The transient complex is formed from the Michaelis complex at a rate (k approximately 3 X 10(-2) S-1) which is some threefold faster than the rate of formation of the irreversibly inactivated complex. The transient complex is, therefore, the principle enzyme form present after short time periods. In the presence of excess clavulanate, however, all the enzyme accumulates into the irreversibly inactivated form. The number of clavulanate turnovers that occur prior to complete enzyme inactivation is 115.


Chemical studies on the inactivation of Escherichia coli RTEM beta-lactamase by clavulanic acid.

Incubation of clavulanic acid with the beta-lactamase from Escherichia coli RTEM leads to enzyme-catalyzed depletion of clavulanic acid, to transient inhibition, and to irreversible inactivation of the enzyme. Both the transiently inhibited and the irreversibly inactivated species show a marked increase in the absorbance at 281 nm that is proportional to the decrease in enzyme activity. Hydroxylamine treatment of irreversibly inactivated enzyme restores about one-third of the catalytic activity, with a concomitant decrease in absorbance at 281 nm. Polyacrylamide isoelectric focusing of the irreversibly inactivated enzyme shows three bands of approximately equal intensity, different from native enzyme. Upon hydroxylamine treatment, one of the three bands disappears and now focuses identically with native enzyme. It is evident that the irreversible inactivation of enzyme by an excess of clavulanic acid generates three products, one of which can be reactivated by hydroxylamine.


Interaction of lipopolysaccharide with detergents and its possible role in the detergent resistance of the outer membrane of Gram-negative bacteria.

In the presence of MgCl2, amounts of detergents which disrupted phospholipid vesicles caused lipopolysaccharide I from Proteus mirabilis to aggregate and form vesicular, membrane-like structures. Vesicle formation with P. mirabilis lipopolysaccharide II containing longer O-polysaccharide chains was extremely poor. Lipopolysaccharides of Salmonella minnesota R mutants (chemotypes Ra, Rc and Re) displayed a growing tendency for vesicle formation with increasing deficiency of the R core polysaccharide. Lipopolysaccharides of chemotypes Rc and Re produced vesicles even in the absence of MgCl2 and detergent. Spherical aggregates consisting of P. mirabilis lipopolysaccharide I MgCl2 and detergent were unable to either entrap or retain [14C]-sucrose, [3H=inulin or [3H]dextran. On the other hand, S. minnesota R mutant lipopolysaccharides of chemotypes Rc and Re could entrap all three saccharides and retain them for at least short periods of time. Leakage of [3H]-inulin out of re-lipopolysaccharide vesicles was greatly retarded by addition of MgCl2 to the vesicle system. Incorporation of P. mirabilis lipopolysaccharide I or S. minnesota Rc lipopolysaccharide into phospholipid vesicles protected these model membranes from disruption by detergent. This suggested a similar protective function of lipopolysaccharide in the outer membrane of enteric bacteria against the action of surfactants occurring in their normal intestinal habitat.


Effects of purine riboside on nucleic acid synthesis in ascites cells.

Purine riboside (nebularine, 9-beta-ribofuranosylpurine) is a naturally occurring base analog which closely resembles adenosine. It inhibits carcinogenic growth. Purine riboside strongly inhibits RNA and DNA synthesis in different cancer ascites cells. Gel electrophoretic analysis of RNA synthesis in vivo in the presence of purine riboside shows the ribosomal components to be inhibited the most. A method for assaying purine riboside or its phosphates intracellularly has been devised, and by using this it has been shown that purine riboside is extensively phosphorylated in the cells. The triphosphate derivative of purine riboside has been isolated and tested in the Escherichia coli RNA polymerase assay. It appears not to be incorporated into this type of RNA and to competitively inhibit this reaction with regard to ATP.


Isolation and characterization of nascent albuminyl peptides synthesized in vivo from rat liver.

A procedure is described for the isolation of nascent albuminyl peptides from rat liver polysomes which is based on the isolation of total peptidyl tRNA by ion-exchange chromatography on ECTEOLA cellulose followed by immuno-affinity chromatography employing monospecific anti-albumin antibodies immobilized on Sepharose 4B. Identity of the isolated nascent albuminyl peptides was assayed by tryptic peptide fingerprint analysis. Quantitation and determination of the specific activity of the nascent albuminyl peptides, labeled in vivo with l-[14c]leucine, were made by subjecting the peptides to acid hydrolysis, dansylation and resoultion of the amino acids by thin-layer chromatography, and determination of the specific activity of dansyl leucine.


A small angle X-ray study of the elongation factor complex Tu . GDP from Escherichia coli.

The protein elongation factor complex Tu. GDP from Escherichia coli was investigated in the presence of 0.01 mM GDP using the small-angle X-ray method. The overall shape and the molecular parameters of the Tu . GDP complex were determined using a least-squares method where the experimental data were used directly without desmearing. The best fit to the experimental data was obtained assuming the molecule to be an ellipsoid of revolution with the semiaxes A = B = 4.08 nm, and C = 1.18nm. Determination of the molecular weight gave the result Mr = 46 000, which corresponds to a water content equal to 26% (by weight).


Comparison of a nitrate reductase-inactivating enzyme from the maize root with a protease form yeast which inactivates tryptophan synthase.

A maize root fraction which inactivates nitrate reductase has been shown to have protease activity which can be measured by the hydrolysis of azocasein. This inactivating enzyme was also found to inactivate yeast tryptophan synthase. Yeast proteases A and B, which inactivate this latter enzyme, also gave a specific inactivation of the maize nitrate reductase. The maize root inactivating enzyme, like yeast protease B, degraded casein, and was inhibited by phenylmethylsulphonyl fluoride. A partially-purified yeast inhibitor prevented catalysis by the yeast proteases and maize root inactivating enzyme, but purified yeast inhibitors were without effect on the latter protein. The level of nitrate reductase-inactivating activity, and associated azocasein-degrading activity, increased with age of the maize root. Evidence was obtained for a heat stable inhibitor which maintained them in an inactive state, especially in the young root tip cells.


Azasterol inhibitors in yeast. Inhibition of the 24-methylene sterol delta24(28)-reductase and delta24-sterol methyltransferase of Saccharomyces cerevisiae by 23-azacholesterol.

The effects of 23-azacholesterol on sterol biosynthesis and growth of Saccharomyces cervisiae were examined. In the presence of 0.2, 0.5, and 1 micron 23-azacholesterol, aerobically-growing yeast produced a nearly constant amount of ergosta-5,7,22,24(28)-tetraenol (approx. 36% of total sterol) and slowly accumulated zymosterol with a concommitant decline in ergosterol synthesis. Growth and total sterol content of yeast cultures treated with 0.2-1 micron 23-azacholesterol were similar to that of the control culture. Yeast cultures treated with 5 and 10 micron 23-azacholesterol produced mostly zymosterol (58-61% of total sterol), while ergosta-5,7,22,24(28)-tetraenol production declined to less than 10% of total sterol. The observed changes in the distribution of sterols in treated cultures are consistent with inhibition of 24-methylene sterol 24(28)-sterol reductase (total inhibition at 1 micron 23-azacholesterol) and of 24-sterol methyltransferase (71% inhibition at 10 micron 23-azacholesterol). Yeast cultures treated with 10 micron 23-azacholesterol were found to contain 4,4-dimethylcholesta-8,14,24-trienol and 4alpha-methylcholesta-8,14,24-trienol, which were isolated and characterized for the first time.


Phospholipid biosynthesis and secretion by a cell line (A549) which resembles type II aleveolar epithelial cells.

The A549 cell line is a continuous cell line derived from a human adenocarcinoma of the lung. At low cell population density the cells contain relatively few lamellar bodies, but in mature cells in very confluent cultures lamellar bodies are abundant. The lamellar bodies from these cells are enriched for phosphatidylcholine and disaturated phosphatidylcholine. In mature cells, 45% of newly synthesized phosphatidylcholine is disaturated. Stimulation with the calcium ionophore A23187 produces exocytosis of phosphatidylcholine (46% disaturated). The A549 cell synthesizes, stores in lamellar bodies, and secretes phosphatidylcholine, and thus has many important biological properties of the alveolar epithelial type II cell.


New procedure for concentration and analytical isoelectric focusing of proteins.

Proteins in aqueous salt solutions (up to 4 mil) were adsorbed by hydrophobic interaction on phenyl-Sepharose gel (0.1 ml) in small columns. After washing out excess salt, gels were applied on the surface of flat bed polyacrylamide gels for isoelectric focusing, which resulted in efficient desorption and transport of protein out of the phenyl-Sepharose gel. There was no difficulty in obtaining a fifty-fold concentration. The following parameters at adsorption of protein were studied: (i) salt concentration in the protein solution; (ii) phenyl-Sepharose gel adsorptive capacity for protein; (iii) suitable volume of washing solution for the phenyl-Sepharose gel. Theoretical aspects on factors promoting adsorption and desorption of proteins on phenyl-Sepharose gel are discussed. Also discussed are earlier used procedures for concentration and/or dialysis. When dilute protein solutions are to be examined for analytical purposes, the proposed procedures seems to be a valuable aid, which does not require expensive equipment, and is quick and simple to perform.


The differential enzyme susceptibility of bovine immunoglobulin G1 and immunoglobulin G2 to pepsin and papain.

Purified bovine immunoglobulins IgG1 and IgG2 were subjected to enzymatic degradation with pepsin and papain. Results were monitored using density gradient ultracentrifugation, acrylamide electrophosesis and immunodiffusion employing subclass- and light chain-specific antisera. The results indicated a marked enzymatic susceptibility of IgG1 to digestion with pepsin. This differential susceptibility can also be demonstrated in unfractionated bovine gamma-globulin. No striking differences between the two subclasses were observed during treatment with papain in the presence of cysteine and after 24 h, most IgG1 and IgG2 was degraded to Fc and Fab fragments. The pepsin Fc fragment generated from IgG2 was larger than that generated from IgG1 although the F(ab')2 fragments were simialr in size. These results are consistent with the hypothesis that the Fc region of IgG1 contains multiple cleavage sites for pepsin whereas IgG2 has few. Rabbits immunized with the first elution peak from a 30 h pepsin digest of bovine gamma-globulin fractionated on Sephadex G-150, responded primarily to common gamma-chain and IgG2-specific determinants. Thus, the differential susceptibility of bovine subclasses to pepsin provides a method for stimulating IgG2-specific antibodies in rabbits.


A mutant of Escherichia coli which accumulates large amounts of coproporphyrin.

A mutant of Escherichia coli which accumulates a large amount of coproporphyrin, presumably because of a block in heme biosynthesis, has been isolated after nitrosoguanidine mutagenesis. On rich media, the mutant forms colonies which give bright orange fluorescence when illuminated with ultraviolet light. The mutant appears to be similar to a Salmonella typhimurium mutant, deficient in uroporphyrinogen III cosynthase, described by Sasarman and Desrochers ((1976) J. Bacteriol. 128, 717--721). A striking property of the mutant is that coproporphyrin is retained within the cells in rich media but is almost totally excreted out of cells in minimal glucose medium.


Calcium and pancreatic beta-cell function. 4. Evidence that glucose and phosphate stimulate calcium-45 incorporation into different intracellular pools.

beta-Cell-rich pancreatic islets were microdissected from ob/ob-mice and used for studies of 45Ca uptake and washout. Irrespective of whether the experiments were performed at 21 or 37 degrees C both glucose and phosphate stimulated the net uptake of lanthanum-nondisplaceable 45Ca. The stimulatory effect of phosphate was additive to that produced by glucose. 45Ca incorporated in response to phosphate differed from that taken up in the presence of 20 mM glucose in being easily washed out although it was not affected by the glucose concentration of the washing medium. The efflux of 45Ca was reduced after introducing phosphate into a medium used to perifuse islets which had accumulated 45Ca in response to 20 mM glucose. This suggests that the outward calcium transport can be influenced also by intracellular trapping of the cation. The glucose-stimulated insulin release was inhibited by phosphate; an effect reversed by the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine. It is concluded that a common effect of glucose and phosphate is to trap calcium in the pancreatic beta-cells but that there are fundamental differences between their effects on intracellular distribution of calcium and on insulin release.


A rapid high-yield purification procedure for the cyclic adenosine 3',5'-monophosphate receptor protein from Escherichia coli.

A new method for the purification of the cyclic adenosine 3',5'-monophosphate receptor protein from Escherichia coli has been developed. The method is rapid and simple, and a very high yield of the purified protein is obtained which is not contaminated with either DNAase or RNAase activity.


Maximum entropy constrained balance randomization for clinical trials.

A specialization of the biased coin clinical trial randomization schemes of Pocock and Simon (1975) and Efron (1971) is proposed which gives guidelines for the coin biasing probabilities of assignment to treatment. The scheme parameterizes the often conflicting goals of random treatment assignment and allocation for stratum balance in terms of a single trade-off parameter specifying a desired degree of determinism. Treatment radomization probabilities are obtained iteratively in an exponential form which maximizes randomization entropy subject to a specified balance constraint.


An application of multivariate ratio methods for the analysis of a longitudinal clinical trial with missing data.

This paper presents an analysis of a longitudinal multi-center clinical trial with missing data. It illustrates the application, the appropriateness, and the limitations of a straightforward ratio estimation procedure for dealing with multivariate situations in which missing data occur at random and with small probability. The parameter estimates are computed via matrix operators such as those used for the generalized least squares analysis of catetorical data. Thus, the estimates may be conveniently analyzed by asymptotic regression methods within the same computer program which computes the estimates, provided that the sample size is sufficiently computer program which computes the estimates, provided that the sample size is sufficiently large.


Combined therapy of human interferon (HI) and secretory immunoglobulin (S-IgA) in the treatment of human herpetic keratitis.

The topical action of a combined therapy of human interferon (3000 U/ml) and secretory immunoglobulin IgA (1,5 mg/ml) was studied in 56 patients with herpetic keratitis. The pain and photophobia disappeared within 48 h after the beginning of treatment and a marked reduction of the corneal lesion during the first week of treatment was observed in all the patients. The therapy was effective, with complete healing of the lesion in 94.8% of cases; 72.2% of them healed in less than 15 days. The highest frequency of healing was between 5 and 10 days, and the rest up to 30 days. Humoral, immunological and delayed hypersensitivity studies were carried out in 36 patients.


Tubular basement membrane deposits in human renal allografts.

In 18 patients who received renal allografts, tubulo-interstitial lesions were studied, according to the types of rejection or the possible recurrence of primary disease. Immunofluorescent (IF) investigation revealed tubular basement membrane (TBM) deposits of IgG and/or C3 and sometimes other components of Complement. The pattern of these deposits was: "linear" in 3 cases, "granular" in 9 cases and "atypical" with linear and granular segments in 6 cases. The IF study of own kidneys of 3 patients, before the transplantation, revealed no TBM deposits. TBM deposits were accompanied by arterial wall deposits of IgM and/or C3 and sometimes C5 in 17 cases. Electron-microscopy study of renal tubules showed thickening, lamellation and/or deposits along the tubular basement membrane. The immunopathogenesis of these deposits was discussed.


The bacteriology of cavitating pulmonary infections and empysema. Studies of transtracheal aspirates and pleural fluid.

A prospective study of bacteriology in 14 cases of cavitating pulmonary infections and empyema is reported. Bacteriologic results were based on transtracheal aspirates or pleural fluid. Appropriate anaerobic bacteriologic methods were employed. Anaerobic bacteria were recovered in 11 patients (79%); they were the only pathogens isolated in 6 patients. The predominant species were Fusobacterium nucleatum, Bacteroides melaninogenicus, Bacteroides fragilis and Peptostreptococcus. Aerobic bacteria were present in 7 patients. The results indicate that anaerobes play a key role in most cases of cavitating pulmonary infections and empyema, and that proper culture of adequat specimens will generally establish the bacteriological diagnosis.


Effect of a long term cholinergic therapy on the gastric acid secretion and on the calculated acid concentration of the parietal component in hypochlorhydric patients.

Fourteen hypochlorhydric patients suffering from atrophic gastritis were tested with pentagastrin and randomly distributed in groups treated per os : one by a placebo, the other by eserine amine oxide. In the placebo group we did not observe a variation in the acid output nor in the acid concentration of the parietal component. In the patients treated with eserine amine oxide, the acid secretion and the acid concentration of the parietal component increased on treatment (p less than 0.01). In two other groups of hypochlorhydric patients tested with pentagastrin (n = 25) or with histamine ( n = 38) and treated for periods of one to three months we also observed an increase in the acid output and the acid concentration of the parietal component. These facts could be explained by an increase in the number of functional cell units, accompanied by a qualitative variation in the parietal component during treatment the patients suffering from atrophic gastritis.


Histo-immunology of myelofibrosis: a study of 100 cases.

A search for immunoglobulin deposits in normal and fibrous interstitial tissue was carried out on 100 bone marrow biopsies (44 myeloproliferative disorders and 56 other hemopathies). This preliminary study enabled one to draw three conslusions: 1) The search for immunoglobulins is only possible on non-decalcified bone marrow. The direct immunofluorescent technique gives better results than peroxidase staining: 2) In myeloproliferative disorders immunoglobulins (especially G) are more frequent and more often associated with a myelofibrosis than in other hemopathies, and 3) The more pronounced the fibrosis the more frequent the immunoglobulin deposits, except at the osteomyelosclerosis stage. They are never found in the fibrosis secondary to intramedullary carcinomatous metastases.


Recent advances in the study of membrane bound saccharides.

The review discusses with the biosynthesis, the changes upon cell density or transformation, the external exposure and the physiological functions of the saccharide moieties of membrane bound sphingoglycolipids and glycoproteins. The structure of biological membranes and some cellular recognition functions tentatively ascribed to membrane bound glycosyl transferases are also reviewed.


Induced radioresistance in four strains of Escherichia coli, two with lambda lysogens.

Cells of E. coli that are recA+ and lex+ show a phenomenon of induced radioresistance. A preexposure to ultraviolet light, or ionizing radiation followed by incubation to allow protein synthesis, followed by treatment with rifampin to prevent further induction, renders the cells resistant to further doses of radiation. When this is attempted with lambda lysogens of the same strains, no radioresistance is seen, even though the preexposure is too small to induce lambda itself. If the lysogens are ind-, namely lambda C1857, about the normal radioresistance can be developed by pretreatment. These findings suggest that the lambda repressors can bind to single-strand breaks caused by the inducing agent and can modify the course of induction.


A means for orienting flat cells in flow systems.

Flattened cells, such as red blood cells, epithelial cells, and sperm of many species, cause problems for fluorescence-activated cell analysis and sorting machines because the flow systems of such devices are unable to control the orientation of these cells as they flow past the detectors. For this reason, the fluorescence or scattered light measurements for identical cells may vary greatly. A flow geometry is here described that orients flat cells in a coaxial flow system so that each cell presents the same aspect to the observation device. A wedge-shaped exit on the sample injection tube in a coaxial flow system is sufficient to produce the desired orientation effect when used with low sample flow rates. Data is presented showing the effect of orientation of fixed chicken erythrocytes on histograms of small forward-angle light-scattering measurements.


Quasi-elastic light scattering from migrating chemotactic bands of Escherichia coli.

We report the observation of migrating chemotactic bands of Escherichia coli in a buffer solution. The temporal development of the bacterial density profile is observed by the scattered light intensity as the band migrates through a stationary laser beam. We have made  a preliminary analysis of the observed band profile with help of the Keller-Segel theory. The model accounts for only some aspects of the observed time evolution of the density profile. The microscopic motility characteristics of the E. coli in the band are simultaneously studied by photon correlation. The measured correlation functions are analyzed to obtain the spatial dependence of the half-width within the band. A simple analytical model is proposed to account for the contribution of the twiddle motion to the correlation function. By analyzing the correlation function as a superposition of straight-line and twiddle motions, we obtain a satisfactory agreement between the theory and the measured angular dependence of the line shape. As a consequence we are able to extract a parameter beta, which measures the average fraction of twiddling bacteria in the center of the band at a given time.


Isolation and physical studies of the intact supercoiled, the open circular and the linear forms of ColE1-plasmid DNA.

For the study of DNA conformations, conformational transitions, and DNA-protein interactions, covalently closed supercoiled ColE1-plasmid DNA has been purified from cultures of Escherichia coli harboring this plasmid and grown in the presence of chloramphenicol according to the method of D.B. Clewell [J. Bact. 110 (1972)667]. The open circular and linear forms of the plasmid were prepared by digestion of the covalently closed, supercoiled form with pancreatic deoxyribonuclease and EcoRI-restriction endonuclease, respectively. The linear form was found to be very homogeneous by electron microscopy and sedimenting boundary analysis. Its physical properties (s0 20,w=16.3 S,D0 20,W=1.98 X 10(-8) cm2 s-1 and [eta]=2605 ml g-1) have been carefully determined in 0.2 M NaCl, 0.002 M NaPO4 pH 7.0,0.002 M EDTA, at 23 degrees C. Combination of s0 20, w (obtained by quasielastic laser light scattering) gave Ms,D=4.39 x 10(6). This value is in reasonable agreement with the molecular weight from total intensity laser light scattering M=4.30 x 10(6). The covalently closed and open circular forms of the ColE1-plasmid are less homogeneous due to slight cross-contamination and the presence of small amounts of dimers in these preparations. The weight fractions of the various components as determined by boundary analysis or electron microscopy are given together with the average quantities obtained in the same solvent for the supercoiled form ((s0 20,w)w=25.4 S, (D0 20,w)z=2.89 x 10(-8) cm2 s-1, [eta]= 788 ML G-1,Ms,D=4.69 x 10(6) and Mw=4.59 x 10(6)) and the open circular form (s0 20, w)w=20.1 S, (D0 20,w)z=2.45 x 10(-8) cm2 s-1, [eta]=1421 ml g-1,Ms,D=4.37 x 10(6) and Mw=4.15 x 10(6)). Midpoint analysis of the sedimenting boundaries allows unambiguous determination of the sedimentation coefficients of these two forms: s0 20,w=24.5 S and s0 20,w=18.8 S, respectively. Also deduced from total intensity light scattering were radii of gyration Rg (103.5, 134.2 and 186 nm) and second virial coefficients A2 (0.7, 4.8 AND 5.4 x 10(-4) mole ml/g2) for the supercoiled, the open circular and linear forms, respectively. The data presented are discussed in relation to the conformational parameters for the three forms in solution.


[Dynamics of insulin secretion in dogs with alloxan diabetes].

It was shown that hypoinsulinemia in the superior pancratico-duodenal vein and inhibition of the first phase of insulin secretion by the pancreas characteristic of alloxan diabetes of different severity was absent in the femoral vein. The data obtained in the insulin and glucose concentration study in the superior pancreatico-duodenal vein stressed the leading role of the pancreatic factor in the pathogenesis of alloxan diabetes in dogs. Analysis of these parameters in the peripheral femoral vein failed to reflect this statement adequately.


[High-frequency generator for the electrocoagulation of the brain tissue].

The authors present a wiring diagram of a high frequency generator intended for coagulation of animal tissue. Stabilization of output current and automatic formation of gradual increase, time delay and decrease of load current permits its employment for stereotaxic decerebration of animals with a good recurrence of the results of coagulation and least traumatic effect.


[Routes of E. coli 055 bacterial penetration through the intestinal wall in gnotobiotic and ordinary animals].

Peculiarities of permeability of intestinal barrier in germfree animals for enteropathogenic E. coli 055 were studied. Both germfree and conventional guinea pigs and rats were used. An increase of bacteriemia was revealed in gnotobiotes during the first day of oral E. coli 055 monocontamination; bacteriemia was transient in conventional animals. Under electron microscopy alterations of intercellular contacts and formation of spaces between enterocytes containing numerous microorganisms were found in the intestinal mucous membrane in gnotobiotes. Also more pronounced changes of microvessels of the intestinal mucous membrane were discovered in gnotobiotes. The processes of ingestionan and digestion of Escherichia coli by enterocytes and leukocytes were noted in conventional animals. The revealed derangements of the intestinal barrier in gnotobiotes explain the cause of higher bacteriemia in germfree animals. An important role of the microbial factor in the formation of intestinal barrier is indicated by the data obtained.


[Effect of ionophore A23187 on the strength of contraction and transmembrane action potential of guinea pig papillary muscle].

Calcium ionophore A23187 being added at 2.5 mM concentration induced 2--3-fold increase of peak contractile force of isolated guinea pig papillary muscle stimulated with the frequency of 0.2 Hz. The increase of force of contraction is not followed by any change in amplitude and duration of membrane action potential. A23187 decreases testing tension as well as duration of the twitch and increases relaxation rate of the muscle. This phenomenon probably reflects increased capability of sarcoplasmic reticulum in the process of calcium ion accumulation. In the presence of ionophore appearance of shoulder on the rising part of the twitch has been registered. In this case the single twitch being differentiated reveals two positive maxima. Addition of D-600 leads to decrease of action potential duration and eliminates the second part of the twitch, the amplitude and the velocity of the first phase of contraction being without change. It was proposed that the first component of the twitch appearing in the presence of A23187 and insensitive to D-600 corresponds to the release of calcium from some intracellular stores.


[Effect of stromal fibroblasts on antibody formation in cultures deficient in A-cells].

Stromal fibroblasts from the monolayer cultures of the rabbit thymus, spleen, and bone marrow when added to suspension cultures of non-adherent rabbit spleen cells had a significant effect on the response of the plaque-forming cells (PFC). Thymus-derived stromal fibroblasts caused an increase in the number of PFC, and bone marrow stromal fibroblasts suppressed the antibody formation in these cultures. Spleen-derived stromal fibroblasts influenced the PFC response similarly to adherent cells. Thymus-derived stromal fibroblasts irradiated by 5000 R influenced the PFC response in the same way as non-irradiated cells. The action of stromal fibroblasts on the antibody formation in the cultures depended on their attachment to the culture flask surface.


[Characteristics of T-cells synthesizing macrophage migration inhibition factor in an H-2 system].

Migration inhibition factor (MIF) was found to be produced by T-cells rather than B-cells as shown by the indirect macrophage migration inhibition test in the H-2 system performed with fractionated lymphocytes. MIF-producers appeared to be more sensitive than T-killer cells to the cytotoxic action of anti-theta-antibodies, plus complement. MIF synthesis by lymph node cells developed earlier after immunization than the cytotoxic activity of these cells. These findings indicated that T-cell subpopulation producing MIF differed from the cytotoxic T-lymphocytes.


[Lymphocyte reactions at early periods after heart transplantation].

The authors determined the quantitative and qualitative composition of the white blood and dehydrogenase activity of lymphocytes in the blood from the afferent and afferent vessels of the transplanted heart at the early postoperative periods in dogs. There was noted a reduction of the lymphocyte count in the blood flowing from the transplanted heart on account of retention of these cells in the transplant, and also an inversion of the mitochondrial dehydrogenases activity due to the lymphocyte contact with foreign antigens of the transplanted heart. A hypothesis is put forward that the inversion mechanism was connected with the lymphocyte functions in the capacity of immunocompetent cells.


[Bound class M, A and G immunoglobulins in the thymus of myasthenia gravis patients].

Deposits of granular material containing IgM, IgA, and IgG were revealed in the thymus of patients with myasthenia by direct immunofluorescence. Treatment of the thymus sections by unlabeled preparations against individual classes of human immunoglobulins inhibited the reaction of the granular material with the homologous labeled preparations. Disappearance of fluorescence of these deposits was also seen in treatment of the sections with glycine-HCl-buffer, pH 2.8. These data permitted a suggestion that granular material represented immune complexes where IgM, IgA, and IgG served as antibody, and thymus tissue components--as an antigen. The presence of bound immunoglobulin in the thymus indicated that an autoimmune process directed against the tissues of this organ occurred in myasthenia.


[Role of normal microflora in the treatment of vibrio carrier state in rat gnotobionts].

Materials on the study of interactions between Vibrio cholera El Tor and some indigenous bacteria of human intestinal tract, which proved to be active antagonists in vitro, are presented in this work. Experiments were conducted on gnotobiotic rats (Fisher), using various combinations and variants of the order of the microorganism administration; there were revealed no antagonistic relations between Virbrio cholerae El Tor and Escherichia coli M-17, Lactobacillus fermenti, Lactobacillus plantarum. Vibrio cholera El Tor disappeared completely from the intestinal tract of the rats after its contact with the fecal microflora of healthy rats.


[Dynamics of formation of antibodies of different immunochemical types to several L-forms and mycoplasmae].

Both IgM- and IgG-immunoglobulins were revealed in the sera of rabbits immunized with L-forms of hemolytic streptococcus, group A, and Mycoplasma fermentans. In the process of immune response the hemagglutinin titres proved to increase, with IgG antibodies prevalence in the hyperimmune sera.


[Compatibility of the F-like plasmid FB1drd with standard F-group plasmids in E. coli K12 strains].

Compatibility of depressed F-like plasmid FB1drd integrated into E. coli K12 cell chromosome with standard plasmids of FI-FVI compatibility groups was studied. The results obtained show that such plasmids can stably coexist in the same cell with plasmid FB1drd. On these grounds it is supposed that the latter belongs to the new compatibility group (FVII).


[Interrelationships in experiments in vitro between the migration activity of leukocytes and RNA and DNA synthesis].

The following correlations were revealed in the parallel study of leukocyte migration in vitro in the presence of a specific antigen and of spontaneous RNA and DNA synthesis in the cultured lymphocytes: 1) a direct correlation between the RNA and DNA synthesis in lymphocytes; 2) a close correlation between the antigen-induced migration and the levels of RNA and DNA synthesis. The effect of the antigen was evidenced by the inhibition or stimulation of leukocyte migration. A high ratio of RNA synthesis to DNA synthesis corresponded to the migration inhibition and a low one--to the migration stimulation. The ratio value varied mainly on account of the changes in the level of DNA synthesis. Participation of T and B cells in the regulation of the antigen-induced leukocyte mobility is discussed.


[Lysosome activity in mice during rejection of a heterotopic heart implant].

Allotransplantation of the heart from newborn mice C57B1/6 into the pinna of adult CBA mice on the 3rd day after operation led to increased acid phosphatase activity and activation of the Golgi apparatus. Numerous lysosomes whose membrane became disrupted on the 7th-9th day from the time of implantation appeared later. At the same time histochemical study demonstrated local loss of acid phosphatase activity and a diffuse staining (brown) of the cytoplasm. No destruction of the lysosome membranes or release of the enzymes into the cytoplasm occurred in case of heart isografts (CBA-GBA).


Leukemic relapse 5 1/2 years after allogeneic bone marrow transplantation.

A 13-yr-old male with acute myelogenous leukemia was treated with various chemotherapy regimens for 3 1/2 yr and then underwent an allogeneic bone marrow transplantation. The donor marrow was successfully engrafted, and the patient remained in remission free of all chemotherapy. Then, 5 1/2 yr later, he developed an extramedullary relapse with a chloroma of his maxillary sinus. This case illustrates the need for prolonged followup of transplant recipients and suggest that statements proposing cure as a result of this procedure may be premature.


Decrease of mast cells in W/Wv mice and their increase by bone marrow transplantation.

Production of tissue mast cells was evaluated in genetically anemic mice of W/Wv genotype and was found to be abnormal. In the skin of adult W/Wv mice the number of mast cells/cm was less than 1% of the number observed in the congeneic +/+ mice. No mast cells were detectable in other tissues of the W/Wv mice. After transplantation of bone marrow cells from +/+ mice the number of mast cells in the skin, stomach, caecum, and mesentery of the W/Wv mice increased to levels similar to those of the +/+ mice. These results show that the W/Wv mouse is a useful tool for the investigations concerning the physiologic roles and the origin of mast cells.


[Experimental models: nutrition and intestinal cancers].

Experimental models of colonic carcinoma chemically induced in rats explored the effects of several factors involved in colon carcinogenesis: high fat level of the diet, quality and quantity of fat, fecal excretion of neutral and acid steroids, intestinal microflora and fiber content of the diet. They suggested an association linking colon cancer to dietary fat and fecal acids and neutral sterols.


[Carcinogens in food (author's transl)].

There is no evidence at present that certain chemicals are responsible for certain human cancers (with the exception of aflatoxin, which is the most suspect agent in the development of primary liver cancer in some districts of Africa and in Thailand). A number of chemicals which are carcinogenic to animals are present in human food. Any chemical which induces tumours in animals should be considered as a potential human carcinogen. But special studies must be undertaken before the role played by these substances in the development of human cancer can be definitely evaluated. First priority should be given to polycyclic aromatic hydrocarbons and N-nitroso compounds by reason of their general distribution in the human environment and their high efficiency in animal experiments. An appropriate approach to such studies would be the establishment of correlations between cancer morbidity data and the data on levels of these chemicals in special environments and, if possible, their total intake. This approach has proved fruitful in the aflatoxin intake studies carried out in countries where liver cancer is endemic. The IARC is currently carrying out a study of this kind in areas of high oesophageal cancer morbidity in Iran and in the north of France.


The study of auditory deprivation from birth.

This paper reviews the research published on auditory deprivation from birth. The extent of the damage caused to animals by this is discussed in relation to humans, deprived of hearing at birth, but later provided with some auditory input through a fitted hearing aid. Lack of research findings prevents a conclusive statement but there appear to be strong grounds for expecting some resulting permanent deficiency at a cortical level in the hearing of complex sounds such as speech.


[Urinary LHRH: radioimmunological assay and physiological significance].

A LHRH-like substance is detectable in the urine. It is extractable by glass powder, which suggests that it is a peptide, and it can be concentrated by anti-LHRH affinity chromatography, strongly suggesting that it is not a contaminant in the RIA. It is composed of very little intact LHRH, and the great majority appears as multiple smaller fractions, although the possibility of "urinary LHRH" being a slightly larger peptide containing a sequence similar or identical to that of LHRH has not been entirely eliminated. Urinary measurements show no correlation with the clinical state, and the widely differing values obtained using different antisera in the present study emphasize the need for in-depth antiserum binding studies prior to clinical use in the LHRH radioimmunoassay.


[Stimulation of growth hormone secretion: a novel extrapigmentary action of alpha-melanotropin].

Alpha-melanotropin stimulated the spontaneous secretion of human growth hormone in vivo and in vitro. This novel extrapigmentary action of alpha-MSH is also demonstrated in preparations of isolated pituitary tissue and cell cultures. It is concluded that alpha-MSH meets some of the criteria of a genuine growth hormone releasing hormone.


A study on socio-medical variables of drug-dependent persons volunteering for treatment in Penang, Malaysia.

There has been a rapid increase in the past five years in the numbers of hospitalized drug dependents. While the studied population was clearly unrepresentative of the country-wide drug using population, it illustrates how the problem is neither limited to one particular stratum of society, nor to the few "traditional" drugs. Indeed, an increasingly youthful group of individuals drawn from all backgrounds is not only becoming dependent upon opiates, but is also using a range of other drugs, all of which are available on the market at relatively low cost. The market prices of drugs have an effect on the pattern of drug use; and many individuals move directly from tobacco to heroin smoking. Drug abuse continues to be a considerable public and governmental concern, and enforcement and treatment programmes are rapidly expanding in attempts to resolve this problem.


Prostacyclin (PGI2) inhibits the formation of platelet thrombi in arterioles and venules of the hamster cheek pouch.

1 Isolated rings of hamster aorta produced an unstable substance which inhibited platelet aggregation in vitro and had the same characteristics as prostacyclin. 2 Prostacyclin inhibited adenosine diphosphate (ADP)-induced aggregation of hamster platelets in vitro. 3 The effects of prostacyclin on ADP-induced platelet thrombi in the microcirculation of the hamster cheek pouch were studied with a television microscope. 4 Prostacyclin caused a dose-dependent increase in the time of iontophoretic application of ADP which was required to induce platelet thrombi formation and embolization in venules (30 to 40 micron diameter). 5 Prostacyclin caused a dose-dependent reduction in the total time during which ADP-induced thrombi were observed following local electrical damage to arterioles (40 to 80 micron diameter). 6 Thrombus formation in venules and arterioles was abolished by 500 ng/ml prostacyclin in the Krebs solution superfusing the hamster cheek pouch. 7 Prostacyclin was approximately twenty times more potent than prostaglandin E1 in preventing thrombus formation in the microcirculation.


Kindling as a model for alcohol withdrawal syndromes.

Periodic brain stimulation, particularly in the limbic system, at stimulus intensities initially too low to produce any behavioural or EEG effects, progressively produces EEG changes, motor automatisms, and eventually convulsions, an effect called kindling. Data are presented and reviewed that suggest that the severity of alcohol withdrawal symptoms progressively increases over years of alcohol abuse in a stepwise fashion similar to the kindling process. The model is presented that the limbic system hyperirritability which accompanies each alcohol withdrawal serves over time to kindle increasingly widespread subcortical structures. These long-term changes in neuronal excitability might relate to the progression of alcohol withdrawal symptoms from tremor to seizures and delirium tremens, as well as the alcoholic personality changes between episodes of withdrawal.


Continuation therapy with amitriptyline in depression.

Thirty-two patients who had responded to amitriptyline (150 mg daily) when suffering from a depressive illness were allocated either to receive placebo or to remain on the same medication for one year. Plasma concentrations of the drug were regularly estimated. There was no correlation between plasma concentration and subsequent residual affective morbidity. In spite of considerable encouragement, three of the patients did not take the prescribed amitriptyline and they all relapsed. Five out of sixteen patients who received placebo relapsed. None of the patients who continued to take amitriptyline relapsed. It is emphasized that the patients studied were selected, inasmuch as they were apparent responders to amitriptyline. It is concluded that this group of patients should continue to be treated with antidepressant medication for eight months after apparent recovery, and care should be taken to ensure the patients' compliance.


Sodium valproate and tardive dyskinesia.

Twenty-five schizophrenic patients with tardive dyskinesia was given 600 mgs sodium valproate daily with their neuroleptic medication. After one month there was no change in their signs, as judged by a panel of nine viewing films of them taken before and at the end of this treatment.


The response of E. coli AB2463 recA to fast neutron beams with mean energies in the range 4 to 27 MeV.

The radiosensitivity of E. coli AB2463 recA, given as the reciprocal of the mean lethal dose, Do-1, has been shown to be the same for four fast neutron beams with widely different energy spectra. It is proposed that this organism can be used to intercompare dosimetry on fast neutron beams with mean energies in the range 4 to 25 MeV with an accuracy of +/- 5%.


Day care after operations for hernia or varicose veins: a controlled trial.

Alternative systems of care after operations for varicose veins or hernia were compared in a total of 360 selected patients, of whom 121 were allocated to be managed in an acute ward for 48 h, 122 in a convalescent hospital for 48 h and 117 to be discharged directly home to the care of the district nursing sister and general practitioner. There were no deaths or major complications. Anaesthetic or surgical problems caused 5 patients (3 convalescent and 2 day care) to be retained in hospital on the day of operation. Minor complications were recorded in approximately one-third of the patients. The majority of these were effectively dealt with by the district nursing sister and only one-third of the complications needed the attention of the general practitioner. Two of the ward patients and 1 of the convalescent patients required readmission to hospital (1 per cent in all). No significant difference was demonstrated in the medical outcome between the three groups after operation. Day care was the most economical of the three systems of care. Inquiry into the patients' opinions elicited the highest proportion of favourable responses in the day care group.


Substance P containing and cholinergic projections from the habenula.

Electrolytic lesions and surgical transection of the habenulo-interpeduncular-ventrotegmental tract have established the existence of separate habenulo-interpeduncular-ventrotegmental substance P and cholinergic projections. Micro-knife lesions separating the habenula nuclei showed the medial habenular nucleus to be the source of substance P fibres running via the fasciculus retroflexus to the ventral tegmental area. The lateral habenular nucleus receives a substance P projection from the medial habenular nucleus and is the source of cholinergic projection to the interpeduncular nucleus and to the medial habenular nucleus. Lesions of the ventrotegmental-interpeduncular area did not modify the levels of substance P and choline acetyltransferase in the habenula. These observations suggested that there are no substance P or ACh containing afferents to the habenula from the ventrotegmental-interpeduncular area and the accumulation of substance P and AChE proximal to but not caudal to transections of the fasciculus retroflexus confirmed this view.


Partial purification of a lipolytic enzyme from Escherichia coli.

An enzyme with phospholipase Al activity was purified some 500-fold from Escherichia coli cell homogenates. Lipase, phospholipase A2, and lysophospholipase copurified with phospholipase A1 and the four activities displayed similar susceptibility to heat treatment. The phospholipase A and lipase activities were recovered in a single band when partially purified preparations were subjected to SDS gel electrophoresis. Phospholipase, lysophospholipase, and lipase all required Ca2+ for activity. Phosphatidylcholine, phosphatidylethanolamine, and their lyso analogues were all hydrolysed at equivalent rates and these were substantially greater than the rate of methylpalmitate or tripalmitoylglycerol hydrolyses under similar incubation conditions. Evidence for a direct but slow hydrolysis of the ester at position 2 of phosphoglyceride was obtained; however, release of fatty acid from this position is mostly indirect involving acyl migration to position 1 and subsequent release of the translocated fatty acid. Escherichia coli, therefore, appears to possess a lipolytic enzyme of broad substrate specificity acting mainly at position 1 but also at position 2 of phosphoglycerides and on triacylglycerols and methyl fatty-acid esters.


The lipase activity of a partially purified lipolytic enzyme of Escherichia coli.

Escherichia coli lipase was found to have a broad pH optimum between pH 8 and 10. Long-chain acyl triacylglycerols such as trioleolglycerol were hydrolysed at a relatively slow rate, whereas, the shorter-chain acyl derivative tracapryloylglycerol was not. Triacylglycerols and diacylglycerols were broken down at a rate 10 to 15 fold greater than that for monoacylglycerol. Simple esters such as methyloleate and cetylpalmitate were hydrolysed at rates greater than that of triacyglycerol. Water-soluble esters such as p-nitrophenylacetate were not attacked. Hydrolysis of lipase substrate occurred more readily in the presence of an anionic detergent such as taurocholate. The enzyme had no marked preference for the 1- or 3-position of triacylglycerols but attacked these positions much more readily than position 2. The enzyme also catalyzed transacylation reaction with simple alcohols such as methanol or ethanol.


relA Gene control of bacterial glycogen synthesis.

Starvation of Escherichia coli K12 for an amino acid results in the stimulation of bacterial glycogen synthesis in cells containing the relA+ gene, but not in cells carrying the relA- allele. Similarly, a large difference in glycogen content is demonstrable between relA+ and relA- cells in stationary phase. It is concluded that guanosine 5',3' -bis(diphosphate) (ppGPP) or some related relA -dependent metabolite is involved in the regulation of bacterial glycogen synthesis. Detection of significant basal levels of glycogen in a relA- strain of E. coli and in unstarved relA+ C. coli indicates that relA control is not absolutely required for glycogen synthesis but serves as a signal for modulation in response to nutrient availability.


The relationship between the spoT gene, the synthesis of stable RNA, ribosomal proteins, and the beta beta' subunits of RNA polymerase following a nutritional shiftup of Escherichia coli.

The level of ppGpp and rates of synthesis of stable RNA, ribosomal protein, and the beta and beta' subunits of RNA polymerase were measured following a nutritional shiftup in Escherichia coli strains, NF 929 (spoT+) and NF 930 (spoT-). In the spoT+ strain, ppGpp levels decreased 50% within 2 min following shiftup, and the rates of synthesis of stable RNA, ribosomal proteins, and the beta and beta' subunits of RNA polymerase increased with little or no lag. In contrast, in the spoT- strain, ppGpp levels transiently increased 40% during the first 6 min following shiftup. An inhibition in the rate of stable RNA synthesis and a delay in the increased synthesis of ribosomal proteins and beta and beta' subunits occurred concurrently with the transient increase in ppGpp. In addition, the DNA-dependent synthesis in vitro of the beta and beta' subunits of RNA polymerase was inhibited by physiological levels of ppGpp. Because of the timing and magnitude of the changes in ppGpp levels in the spoT- strain versus the timing when the new rates of stable RNA, ribosomal protein, and beta and beta' subunits synthesis are reached, it is concluded that ppGpp is not the sole element regulating the expression of these genes.


Structure and function of aspartate transcarbamoylase studied using chymotrypsin as a probe.

Aspartate transcarbamoylase from Escherichia coli is composed of six catalytic (c) and six regulatory (r) polypeptides. We have studied the structure and function of this enzyme using chymotrypsin as a probe. The protease inactivates the isolated catalytic subunit (c3) but has not effects on the native enzyme (c6r6). Under identical conditions, the c3r6 complex is inactivated at a much slower rate than c3. The presence of the substrate analogue succinate together with carbamoyl phosphate reduces substantially the rate of inactivation. Extended exposure to chymotrypsin converts the catalytic subunit into a partially active derivative with a fourfold higher Michaelis constant. This derivative is indistinguishable from the unmodified catalytic subnit in gell electrophoresis under nondenaturing conditions. However, in the presence of sodium dodecyl sulfate, the major fragment in the electropherogram is smaller than that of the intact catalytic polypeptide. The results could be explained by postulating the presence of a chymotrypsin-sensitive peptide bond at or near the active site. Since X-ray crystallographic studies have indicated that the active sites are located in a central cavity, the resistance of the native enzyme towards inactivation may be due to the inability of chymotrypsin to enter this cavity.


Isolation of Ureaplasma from bovine granular vulvitis.

Cultures for mycoplasmatales, viruses and bacteria were made from bovine vulvar swabs to determine whether ureaplasma was associated with a clinical granular vulvitis observed in 16 Ontario dairy herds. Ureaplasma was isolated from 23.5% of 34 clinically normal cows, 74% of 27 cows with mild to moderate vulvar hyperemia but no discharge and 100% of 20 cows with acute vulvar hyperemia accompanied by purulent discharge. There were statistically significant differences in rates of isolation among clinical groups. Mycoplasma bovigenitalium was isolated from 7.7% and 20% of cows with moderate or acute vulvitis respectively but not from normal cows. Haemophilus somnus was isolated from 25% of cows with acute vulvitis. There were no significant differences in isolations of Escherichia coli, Corynebacterium pyogenes and alpha-hemolytic streptococcus between normal and clinically affected animals. Cultures of 135 repeat samples from 33 cows revealed that ureaplasma persisted in some animals for at least three months. No viruses were isolated from any of the animals in this study.


Effect of different treatments on the activity of the heat labile enterotoxin of Escherichia coli F11(P155).

A whole cell lysate preparation of the Escherichia coli strain F11(P155) was treated with the following agents: heat (60 degrees C for 30 minutes), pronase, lipase, amylase, formalin (0.1%), sodium lauryl sulfate (0.05%) and sarkosyl NL30 (0.05%). Except for the amylase treatment, the treated whole cell lysate was inactivated when tested in rabbit gut loops or on Chinese hamster ovary cells. The preparations treated with heat or formalin could be used to produce a neutralizing antiserum or could remove the neutralizing capacity of an anti-F11(P155) serum. Finally, the attempt to demonstrate enterotoxicity with the cytoplasmic membrane of the bacterial cell failed.


Optimal macroculture method for studying mitogenic stimulation of turkey lymphocytes.

This report describes a method for culturing turkey lymphocytes in disposable, unwashed glass test tubes with Morton closures and for recovering lymphocytes on fiber glass filters with a cell harvester made of common laboratory equipment for assay of mitogenic stimulation. Optimal conditions for culture were established.


A radiotracer method for the measurement of central nervous system catecholamines in vivo.

A new strategy for measurement of brain catecholamines was tested in an animal model. [3H]Norepinephrine was infused intravenously in rabbits to label the peripheral norepinephrine pools. The specific activity of urinary 3-methoxy-4-hydroxymandelic acid was consistently higher than that for 3-methoxy-4-hydroxyphenyglycol (MHPG). Central sympathectomy with 6-hydroxydopamine abolished this difference. Using the formula we propose, it is estimated that 30-50% of urinary MHPG originates from the central nervous system.


Evolution of ethylene by Saccharomyces cerevisiae as influenced by the carbon source for growth and the presence of air.

Effects of the carbon source and oxygen on ethylene production by the yeast Saccharomyces cerevisiae have been studied. The amounts of ethylene evolved by the yeast culture were less than those detected in the blank (an equal volume of uninoculated medium), suggesting a net absorption of ethylene by the yeast cells. Addition of glucose to the lactate-grown yeast culture induced ethylene production. This glucose-induced stimulation of ethylene production was inhibited to a great extent by cycloheximide. Results suggested that the yeast cells in the presence of glucose synthesized an ethylene precursor and passed it into the medium. The conversion of this precursor to ethylene might be stimulated by oxygen. The fact that ethylene was produced by the yeast growing anaerobically and also by respiration-deficient mutants isolated from the wild-type yeast suggested that mitochondrial ATP synthesis was not an absolute requirement for ethylene biogenesis.


Detection of Mycoplasma hominis and Mycoplasma orale in cell cultures by immunofluorescence.

Mycoplasma contaminants of animal and human cell cultures were rapidly detected and identified by an indirect immunofluorescent technique. Cells suspected of being contaminated by mycoplasmas were grown as monolayers on chamber slides in a culture medium selected to promote mycoplasmal growth. Before fixation by acetone, the monolayers were subjected to a hypotonic treatment to cause swelling of the mycoplasmas. Detection and identification were then performed by indirect immunofluorescence using rabbit antisera to various mycoplasma species. The correlation between results obtained by the standard isolation procedure and those obtained by this method was very close.


Effect of amino acid deprivation and chloramphenicol treatment on cell sizes of rel+ and relA- strains of Escherichia coli.

The effects of inhibition of protein synthesis on the cell size distributions of rel+ and relA- derivatives of Escherichia coli K-12 were determined. Amino acid deprivation resulted in a reduction in the cell sizes of rel+ strains but not of relA- strains. Treatment with chloramphenicol (CAM) did not alter the size distributions of either rel+ or relA- strains except when they were rel+ dap-. CAM treatment of rel+ dap- strains resulted in an increase in cell size. It is proposed that these results reflect differences in the structures of the cell envelopes of rel+ and relA- bacteria.


Regulation of brain pyruvate dehydrogenase multienzyme complex.

A number of excellent and comprehensive reviews on various aspects of pyruvate dehydrogenase multienzyme complex have been written recently. The purpose of the present review is to summarize briefly the reaction mechanism and the regulation of this enzyme. Emphasis is put on the most recent literature not covered by previous reviews. Particular attention is also paid to the regulation of brain pyruvate dehydrogenase multienzyme complex, since a number of patients with neuromuscular diseases, such as Friedreich's ataxia, show a decreased rate of pyruvate oxidation.


Ex vivo removal of serum IgG in a patient with colon carcinoma: some biochemical, immunological and histological observations.

Plasma of a patient with metastatic colon carcinoma was perfused over Formalin and heat-killed S. aureus, in an extracorporeal filtration apparatus, in order to nonspecifically remove IgG and its complexes. Twenty ex vivo absorption procedures were done, over a five-month period, with a minimum of discomfort to the patient. Extracorporeal perfusion of plasma on S. aureus effectively reduced the levels of IgG and immune complexes in the perfused plasma. The nonspecific removal of IgG resulted in 1) slight biochemical alterations in the serum, 2) a transient reduction in the serum blocking activity and appearance of complement-dependent serum cytotoxicity, 3) an increase in the serum IgM levels, 4) a transient increase in the Ig surface-bearing lymphocytes and a decrease in "E" rosetting lymphocytes in the first 24-48 hours postperfusion, particularly during the early treatments, 5) an improvement in general condition of the patient and decrease in tumor size, and 6) histological changes in the tumor consistent with tumor destruction. The potential problems and clinical applications of procedures involving ex vivo specific or nonspecific immunoabsorbents are discussed.


Randomized phase II clinical trial of adriamycin, methotrexate, and actinomycin-D in advanced measurable pancreatic carcinoma: a Gastrointestinal Tumor Study Group Report.

Sixty-six patients with advanced pancreatic carcinoma were randomized to receive single agent chemotherapy with either adriamycin, methotrexate, or actinomycin-D using conventional dose, route and schedule of administration. All patients had measurable lesions which were used to objective assessment of response. For adriamycin, 2 of 25 patients (8%) evidenced a partial response (2 of 15 (13%) previously untreated patients). One of 25 patients treated with methotrexate and one of 28 received actinomycin-D responded. The duration of responses ranged from 43-64 days for those patients with no chemotherapy prior to study entry. The median survival of patients who received adriamycin as initial treatment was 12 weeks compared to 8 weeks for methotrexate and 6 weeks for actinomycin-D therapy.


Primary malignant teratoma of the thyroid: case report and literature review of cervical teratomas in adults.

A case of primary malignant teratoma of the thyroid in a gravid 37-year-old female is reported and the literature on cervical teratomas in adults reviewed. The patient was treated by total thyroidectomy and left radical neck dissection followed by cervical and mediastinal radiotherapy and chemotherapy. She fied of recurrent tumor and related complications 9 months after discovery of the lesion. This is the 6th case of primary teratoma of the thyroid and the 9th case of cervical teratoma reported in a adult. These tumors are characteristically large, rapidly growing and highly malignant. The longest survival reported is 10 months. The differentiation of this lesion from carcinosarcomas of the thyroid is discussed.


Treatment of histiocytic and mixed lymphomas: a comparison of two, three and four drug chemotherapy.

The Eastern Cooperative Oncology Group has studied 187 patients with generalized progressive malignant lymphoma classified as having the histologic sub-types histiocytic or mixed. Histology review by the Pathology Panel for Lymphoma Clinical Trials demonstrated a 31% disparity with contributing institution's pathologists in regard to cell type, but good agreement with interpretation of nodular or diffuse nodal pattern. Patients were assigned at random to treatment with cyclophosphamide 1 g/m2 on day 1 and prednisone 100 mg/m2 daily for five days (CP); CP plus vincristine 1 mg/m2 on day 1 (CVP); or CVP plus BCNU 60 mg/m2 on day 1 (BCVP). Chemotherapy was given for nine, twenty-one day cycles. The observed complete remission rates were CP-21%, CVP 34%, BCVP 34%. Both CVP and BCVP had significantly more complete remissions than CP, but survival following CVP (118 weeks) was significantly longer than that following either BCVP (76 weeks) or CP (74 weeks). Histologic sub-type, lymph node pattern, response to chemotherapy, performance status and stage of disease were also found to influence survival.


Casein and alpha-lactalbumin detection in breast cancer cells by immunocytochemistry.

Casein and alpha-lactalbumin are characteristic proteins produced by normal mammary cells under hormonal stimulation. Specific antisera were used to immunochemically stain for these two proteins in the unstimulated R3230AC transplantable rat mammary tumor as well as in normal lactating rat mammary tissue. Specific staining was observed in the lactating rat alveolar epithelial cells and in the cells lining the intraalveolar ducts with both antisera. In the R3230AC tumor some cells were shown to contain casein or alpha-lactalbumin; the majority of the tumor cells were unstained. These findings indicate that normal mammary cells as well as a small population of cells within the R3230AC tumor are actively synthesizing casein or alpha-lactalbumin. Furthermore, the results suggest that immunocytochemistry may be used to determine the functional heterogeneity of mammary tumors directly.


Overview of cardiac pathology in relation to anthracycline cardiotoxicity.

A review is presented of cardiac pathologic changes associated with acute and chronic toxicity of anthracyclines. These changes consist of cardiac dilatation, degeneration and atrophy of the muscle cells, and interstitial edema and fibrosis. The degeneration of cardiac muscle cells is a complex phenomenon that involves the myofibrils, the nucleus, the mitochondria and the membrane systems of the T-tubules, the sarcoplasmic reticulum, and the intercellular junctions. The pathogenesis of these alteration is discussed in terms of the biochemical effects of anthracyclines.


Perspective on the physiology of hypertension.

Present understanding of the physiology of arterial pressure regulation indicates that the renal-body fluid volume system determines the level at which the mean pressure resides over long periods of time. The relationships between blood volume, and size and compliance of the entire vascular system, and intrinsic regulation of tissue blood flow determine the sequence of observed changes in cardiac output and total peripheral resistance. Current evidence is compatible with the concept that functional changes in the renal vasculature or tubular system, either intrinsic or extrinsic in origin, reflect the final common pathway in the genesis of all forms of experimental and human hypertension. At the present time the nature of these renal changes appears to alter the fundamental relationships between renal perfusion pressure and sodium and water excretion. One of the major challenges in the field of hypertension today is to test the hypothesis that changes in renal function, either extrinsic or intrinsic in form, are involved in all forms of hypertension.


Perspectives on physiological monitoring: junctional-type potentials in the food ventricle.

1. Many toads monitored throughout survival with no support other than protection against drying, pass terminally through a remarkable evolution which is described here in the full details of a single experiment lasting some 40 hours. 2. The essential features of this particular sequence is block of the Luciani-Wenckebach type affecting SA, AV, and intraventricular conduction. SA block was apparently the major cause of periods of arrest and of cycles of heart beats. Periodically PR delay based on progressive AV block was observed but it was not an outstanding feature. 3. Progressive, rate-determined intraventricular block during the cycles of ventricular beats was the first new feature of these observations. 4. As intraventricular block progressed, an initial ventricular deflection separated itself from the rest of QRS. 5. This initial deflection diminished in amplitude throughout each cycle of ventricular beats, its rate of rise diminished, and the interval separating it from the rest of the ventricular complex increased until the whole initial deflection was revealed. 6. Thereafter, with a small decrease in amplitude of the initial deflection, the remainder of the ventricular electrogram failed to follow and the complex stood alone. 7. Its polarity indicated its origin at the base of the ventricle, the interval separating it from the origin of P indicated that it was downstream from the AV conduction mechanism. 8. This deflection, now a local ventricular potential (LVP) then progressively declined in amplitude and disappeared. 9. The possibility has been discussed that the potential represents (a) a true action potential localized by block or (b) a local, nonpropagated potential akin to junctional potentials like: (1) end-plate potentials, (2) generator potentials, (3) excitatory postsynaptic potentials (EPSPs), or (c) a pacemaker potential. The experiments that have revealed the phenomenon have not provided other than suggestive but inconclusive information about its nature. 10. The observations are new or certainly not well known and further study should shed light on the problem of intracardial impulse formation and conduction.


Localization of the pituitary lactotropes and thyrotropes within Ambystoma gracile by histochemical and immunochemical methods. A developmental study of two populations.

The prolactin-producing cells are the first hormone-producing cells of the pars distalis to be differentiated within Ambystoma gracile. They first appear when the larvae attain a length of approximately 3.0 cm snout to vent length (SVL). Thyrotropes are observed as the next chromophilic cells to appear, and they occur when the larvae are approximately 4.5 cm SVL. Both thyrotropes and lactotropes increase in numbers until metamorphosis. Gonadotropes begin to appear when larvae attain a size of 5.0 cm SVL and become extremely abundant when larvae are in excess of 7.0 cm SVL. Animals, generally, exhibit the greatest number of thyrotropes just prior to the mean size for metamorphosis, and metamorphosing animals exhibit a dramatic reduction in the number of thyrotropes. Neotenous larvae have an abundant number of thyrotropes which are mainly located along the caudal periphery of the pituitary.


B lymphocyte differentiation in lethally irradiated and reconstituted mice. A histological study using immunofluorescent detection of B lymphocytes.

The recovery of the B lymphocyte compartments was investigated in lethally irradiated mice reconstituted with fetal liver cells. This was done by means of immunofluorescence on frozen sections of spleen, lymph nodes and Peyer's patches. The first B lymphocyte recovery in the spleen was observed on day 8, a few days earlier than in lymph nodes and Peyer's patches (day 13). These early B cells in the spleen were found in the central part of the periarteriolar lymphatic sheath (PALS). Later on, while increasing in number, the B cells formed growing follicles at the periphery of the PALS. Subsequently, brightly fluorescent B cells appeared in the marginal zone, which surrounded the follicles. Another two weeks later, around day 30, also germinal center formation was observed in the follicles of the spleen. B cell development in lymph nodes and Peyer's patches started somewhat later than in the spleen, but once started, the recovery of the different compartments was completed very fast. Germinal center reactions were found in lymph nodes and Peyer's patches already on day 25, and thus earlier than in the spleen, but later than the first occurrence of the strongly fluorescent cells in the marginal zone. Apparently, germinalcenter formation is not essential for the recovery of the population of brightly fluorescent B cells in the marginal zone after irradiation and reconstitution.


Shunt quantification by mathematical analysis of indicator dilution curves.

Mathematical models are presented for describing and analyzing indicator dilution curves recorded in patients with intracardiac and great vessel shunts. The models treat individual segments of the circulation as linear system blocks, each having, at its output, a characteristic time response to a rapid injection of indicator at its input. These blocks are combined in feedback and feed-forward configurations to simulate left-to-right, right-to-left, and bidirectional shunts. A shunt analysis algorithm, using discrete analogs of the linear system models, was implemented in a computer program and used to analyze thermodilution curves recorded in patients with congenital heart defects. Results are presented comparing shunt fractions obtained from thermodilution curve analyses with oximetrically determined values in 20 patients. Comparing left-to-right shunts measured by the two methods, the mean systematic difference was 0.7% of pulmonary flow and the standard deviation was 7.6% of pulmonary flow. Statistical validation of the bidirectional shunt method will require acquisition and analysis of more data; however, reasonable shunt fractions were computed in five cases studied and good agreement with oximetric determinations was obtained in two cases where complete oximetric data were available.


Patterns of protein synthesis in E. coli: a catalog of the amount of 140 individual proteins at different growth rates.

The amount of 140 individual proteins of E. coli B/r was measured during balanced growth in five different media. The abundance of each protein was determined from its absolute amount in 14C-glucose-minimal medium and a measurement of its relative amount at each growth rate using a double labeling technique. Separation of the proteins was carried out by two-dimensional gel electrophoresis. This catalog of proteins, combined with 50 additional ribosomal proteins already studied, comprises about 5% of the coding capacity of the genome, but accounts for two thirds of the cell's protein mass. The behavior of most of these proteins could be described by a relatively small number of patterns. 102 of the 140 proteins exhibited nearly linear variations with growth rate. The remaining 38 proteins exhibited levels which seemed to depend more on the chemical nature of the medium than on growth rate. Proteins, including the ribosomal proteins, that increase in amount with increasing growth rate account for 20% of total cell protein by weight during growth on acetate, 32% on glucose-minimal medium and 55% on glucose-rich medium. Proteins with invariant levels in the various media comprise about 4% of the cell's total protein.


Cell surface markers for distinguishing different types of rat dorsal root ganglion cells in culture.

The distribution of three cell surface markers on dissociated cultures of the rat dorsal root ganglion has been investigated using indirect immunofluorescence. Tetanus toxin binds to a nondividing class of cells with neuronal morphology. Ran-1 is expressed by Schwann cells. Thy-1 is expressed both by fibroblasts and by all the neurones. These assignments have been confirmed by detection of any two of these markers in a single culture using antisera conjugated to different fluorochromes. Approximately 20% of the nonneuronal cells in these cultures were negative for all three markers.


A yeast mutant which accumulates precursor tRNAs.

It has been proposed that the conditional yeast mutant ts136 is defective in the transport of mRNA from the nucleus to the cytoplasm (Hutchinson, Hartwell and McLaughlin, 1969). We have examined ts136 to determine whether it is defective in tRNA biosynthesis. At the restrictive temperature, the mutant accumulates twelve new species of RNA. These species co-migrate on polyacrylamide gels with some of the pulse-labeled precursor tRNAs. Three of the new RNAs (species 1a, 1b and 1c are large enough to contain two tandom tRNAs. Although RNAs 1a, 1b, and 1c do not contain detectable levels of modified and methylated bases, at least one of them hybridizes to DNA from an E. coli plasmid containing a yeast tRNA gene. All the remaining RNAs (2--8) contain modified and methylated bases typical of tRNA. Three of these species were tested and were found to hybridize to tRNA genes. Ribosomal RNA synthesis is also defective in ts136. It is suggested that ts136 may be defective in a nucleolytic activity, which is a prerequisite to RNA transport.


Transcription and processing of intervening sequences in yeast tRNA genes.

Genes for yeast tRNATyr and tRNAPhe have been sequenced (Goodman, Olson and Hall, 1977; Valenzuela et al., 1978) which contain additional nucleotides (intervening sequences) within the middle of the gene that are not present in the mature tRNA. We have isolated precursors to rRNATyr and tRNAPhe from a yeast temperature-sensitive mutant (at the rna1 locus) which accumulates only certain precursor tRNAs at the nonpermissive temperature. The tRNATyr and tRNAPhe precursors were analyzed by oligonucleotide mapping; they each contain the intervening sequence and fully matured 5' and 3' termini. Furthermore, these precursors were used as substrates to search for an enzymatic activity which can remove the intervening sequences and religate the ends. We have shown that wild-type yeast contains such an activity, and that this activity specifically removes the intervening sequences to produce mature-sized RNAs.


Biochemical mechanisms accounting for the toxic action of oxygen on living organisms: the key role of superoxide dismutase.

It seems that superoxide dismutase plays the key role in protecting aerobes against O2 toxicity, but there is a whole range of ancillary mechanisms: enzymes to remove H2O2 (catalase, peroxidases) and hence to control formation of .OH from O2, which requires H2O2; antioxidants (ascorbate, GSH, alpha-tocopherol, carotenoids), which also react with singlet oxygen and/or .OH and often inhibit lipid peroxidation and last, but not least in animals, glutathione peroxidase, which controls the rate of lipid peroxidation. These mechanisms cope well at normal O2 concentrations but are insufficient at higher levels.


[A probable laboratory contamination with Plasmodium falciparum].

A student working in a medical biology department presented a severe Plasmodium falciparum malaria, with high thrombopenia, hypercalcemia and hypophosphoremia. The mode of infection seems to be a laboratory contamination with a highly contaminated blood sample through skin excoriations.


[2 cases of Plasmodium falciparum malaria. Probable responsibility of Anopheles transported by air].

Report of two cases of Plasmodium falciparum malaria which seem to be originary from the Pariser area: the first one in a luggage carrier working in an airport; the second one in a subject living in the neighbourhood of this airport. These cases perhaps question the hygiene measures taken on the arrival of planes coming from impalutaded regions.


[Post-transfusion pernicious Plasmodium falciparum attack in a patient with renal insufficiency].

The authors report one case of transfusion induced falciparum cerebral malaria. Among the parasites responsible for transfusion malaria, Plasmodium falciparum is rarely implicated, but may be accompanied by serious complications. Different ways of artificial transmission of the parasites are reported. Clinical findings are few, so the examination of blood films is essential for diagnostic. The dangerous blood donor must be identified by immunofluorescent methods. The specific treatment is quinine for cerebral malaria or chloroquine in other cases.


Comparison of immunofluorescence and immunoperoxidase methods for viral diagnosis at a distance: a WHO collaborative study.

Nasopharyngeal secretions collected in Newcastle were examined in both Newcastle and Stockholm for the presence of influenza virus type A and respiratory syncytial (RS) virus by immunofluorescence and immunoperoxidase techniques. A total of 139 specimens were examined in this way and the agreement between the two centres for immunofluorescence was 94% for influenza virus A, 95% for respiratory syncytial virus, and 95% for negative specimens. This technique can therefore be used for examining specimens taken at great distances from the collecting laboratory. The results of the immunoperoxidase technique were unsatisfactory mainly because of the presence of endogenous peroxidase. Measures taken to remove this also destroyed RS virus antigen. It is premature to introduce the immunoperoxidase technique for rapid virus diagnosis, but in due course, when the problems outlined in this article are solved, it could become a useful technique.


Progress in the rapid diagnosis of viral infections: a memorandum.

This Memorandum reports the progress that has been made over the past year in the development of laboratory techniques for the rapid diagnosis of viral infections. Progress is reported in the use of the immunoperoxidase and immunofluorescence techniques for detecting viruses in respiratory secretions, the use of the glutaraldehyde and periodate methods for peroxidase conjugation, the application of enzyme immunoassay techniques in the detection of viral antigens and antibodies, and training in the use of these techniques. A number of recommendations for further research are made.


Hepatic oxygen supply and plasma lactate and glucose in endotoxic shock.

Hepatic oxygen supply and selected blood parameters were recorded in fasted male rates given 20--30 mg/kg Escherichia coli endotoxin intraperitoneally. Mortality was 70% within 24 hours. Measurements during the initial eight hours postendotoxin recorded no differences of hematocrit, systemic arterial pressure, or arterial pO2 between survivors and eventual nonsurvivors. However, by the sixth or eighth hour nonsurvivors showed significantly higher plasma lactate, lower plasma glucose and blood pH, and a greater degree of hypocapnea. In addition, a mean hepatic pO2 had decreased from 25.2 mm Hg during the control to 3.8 mm Hg after six hours. A decline of hepatic oxygen supply also occurred in surviving rats but was significantly less severe. Control rats showed a mild degree of respiratory alkalosis but were otherwise stable over eight hours. The relationship of hepatic oxygen supply to differences of plasma lactate and glucose is discussed. Failure of hepatic circulation is cited as the probable cause of extensive liver anoxia and related developments in nonsurviving endotoxic rats.


[Comparative value of immunofluorescence and immunoenzymology on figured antigen in the immunological diagnosis of African trypanosomiasis].

Indirect immunoperoxydase method is compared with indirect immunofluorescence using immunized animal and human sera, and negative control sera. The obtained results show a good statisticly established concord and allow to propose the indirect immunoperoxydase method being used in the biological diagnostic of african Trypanosomiasis because its easy carrying out and its examin with an ordinary microscope.


Fibrinolysis and risk factors of atherosclerotic disease, with special emphasis on diabetes mellitus.

This paper reviews and discusses the evidence supporting the involvement of defective fibrinolysis in the pathogenesis of atherosclerosis, with emphasis on diabetes mellitus. According to the literature, defective fibrinolysis has been observed in association with virtually every major "risk factor" for coronary heart disease, including diabetes mellitus, hypercholesterolemia, hypertriglyceridemia, hypertension, obesity, cigarette smoking and lack of physical exercise. The interrelationships between disturbances in carbohydrate and fat metabolism and fibrinolysis are considered. Attention is drawn to the need for increased clinical attention to the potential role of defective fibrinolysis in atherogenesis, and periodic assessments of the fibrinolytic status are suggested as a promising approach toward early recognition of atherosclerotic tendency and risk. The judicious use of physiologic, dietary and pharmacologic means to correct defective fibrinolysis prophylactically and for the treatment of some forms of atherosclerosis is advocated.


Surgery using cardiopulmonary bypass in the elderly.

This study included 89 patients, 70-82 years (mean 72.8 years), who had procedures using cardiopulmonary bypass since 1955. Twenty-six patients had elective aortic valve replacement (AVR), with two hospital deaths. One patient who underwent emergency AVR for bacterial endocarditis died of septic shock. Ten patients had AVR and coronary artery bypass surgery (CABG), with one hospital death (10%). Fourteen patients had mitral valve replacement (MVR), with eight hospital deaths (57%). Two died of left ventricular rupture after leaving the operating room, and the remainder died of low cardiac output. Twenty-five patients had CABG with no early deaths. Seven patients had aneurysms of the thoracic aorta, with two early deaths. Six patients had other procedures with one death, making a total of 16 operative deaths in the 89 patients. Eighty-four of the patients (94%) were New York Heart Association (NYHA) Functional Class III or IV for congestive heart failure and/or angina, preoperatively. Of these, 12 were in extremis immediately before surgery, and six survived. There were 10 late deaths. The actuarial survival rates for one, two and five years for all patients were 69% (40 patients), 47% (20 patients) and 21% (seven patients), respectively. At recent follow-up (mean 20 months) 84% of the hospital survivors were symptomatically improved at least one NYHA Functional Class. We conclude that CABG and/or AVR can be performed in elderly patients with a low hospital mortality and with symptomatic improvement. However, MVR in the elderly carries an unusually high mortality (7.3 times greater than patients less than 70, in our experience), and this risk must be weighed when considering MVR in these patients.


Thermometric enzyme linked immunosorbent assay in continuous flow system: optimization and evaluation using human serum albumin as a model system.

Thermometric enzyme-linked immunosorbent assay (TELISA) is described. After the procedure of optimization, human serum albumin was assayed using anti-human serum albumin bound to Sepharose CL 4-B in the enzyme thermistor unit and catalase as label on the free antigen. The model system was used for assays down to 10(-13)M and the preparation of immobilized antibodies was used repeatedly up to 100 times. Comparative studies of the TELISA technique with bromocresol green, immunoturbidimetric and rocket immunoelectrophoretic methods were carried out and showed that TELISA could be used as an alternative method.


Hypothalamo-pituitary-gonadal function in male central precocious puberty.

Eleven boys aged 1-10 years with central precocious puberty were studied. According to the pubertal development six were classified as P2, one as P3 and four as P5. In all cases plasma testosterone levels were definitely elevated (1.7-5.8 ng/ml) when compared with pre-pubertal controls. Peak values after HCG (3 X 1500 units) in four of the boys were in the high adult range. The binding capacity of serum testosterone oestradiol binding globuline (TeBG) ranged between 0.5 and 7.30 microgram/dl. Basal plasma levels of LH and FSH were respectively 2.06 +/- 0.64 and 1.2 +/- 0.25 miu/ml, and peak levels after LHRH (0.1 mg/m2) 13.9 +/- 3.7 and 2.6 +/- 0.43 miu/ml respectively. The data demonstrated a significant increase of plasma testosterone and post LHRH LH peak levels in boys with central precocious puberty when compared with pre-pubertal controls. The patients at stage P2 exhibited high levels of plasma testosterone contrasting with the degree of pubertal maturation, high values of TeBG and low response to LHRH which were in the pre-pubertal range. These findings suggest that the testicular sensitivity to LH increases early in boys with central precocious puberty, while the testosterone responsiveness, both at peripheral and hypothalamic levels, is delayed.


Traumatic ankylosis of the temporomandibular joint.

Traumatic injuries to the region of the temporomandibular joint (TMJ) may result in varying degrees of hypomobility of the mandible. The physical and psychological problems associated with restricted jaw opening are outlined. The different forms of ankylosis are mentioned. The rationale behind the various surgical approaches to the problem is outlined, and mention is made of current concepts in joint reconstruction. The importance of supportive adjunctive therapy in the postoperative period, is emphasized.


Pompe's disease: diagnosis in kidney and leucocytes using 4-methylumbelliferyl-alpha-D-glucopyranoside.

The diagnosis of Pompe's disease by the assay of acid alpha-glucosidase in kidney and leucocytes was not previously possible because of the presence of another component which had activity at pH 4.0, but was not deficient in the disease. This problem was resolved either by the use of the inhibitors, turanose, maltose and citrate, or by isoelectric precipitation at pH 5.0, which enabled the estimation of acid alpha glucosidase in kidney and leucocytes.