A dnaB analog ban, specified by bacteriophage P1: genetic and physiological evidence for functional analogy and interactions between the two products.

Bacteriophage P1 has been shown previously to determine a product ban that can substitute in DNA replication for the protein specified by cis-tron dnaB of Escherichia coli. However, ban product furnished by P1 bac prophage (ban constitutive) substitutes only poorly for DNA replication in the absence of dnaB product in a strain bearing an unsuppressed amber mutation, dnaB266, as shown by the cryosensitivity of the dnaB266 (P1 bac) lysogen and its unability to support lambda growth. An additional mutation (termed crr) in the P1 bac prophage has been obtained which confers cryoresistance to the sup+ dnaB266 (P1 bac crr) lysogen and restores its ability to support lambda growth. ban product produced in P1 bac crr lysogen fulfills all dnaB roles in vivo, especially in the various instances in which ban product expressed in P1 bac lysogens does not. The ban product is expressed constitutively in P1 crr prophage. The crr-1 mutation is tightly linied to the bac-1 and ban-1 mutations and is dominant over crr+. The nature of the crr mutation is discussed: two hypotheses are considered, that of a mutation in the ban gene rendering the ban product more active or that of a site mutation in the ban operon increasing the level of ban expression. Expression of ban product (wild type or altered) leads to interactions with the variously altered dnaB product. Both positive and negative interactions are described. Genetic results presented here suggest that ban and dnaB subunits interact to form hybrid dnaB-like molecules; the average composition of which depends on the relative quantities of ban and dnaB subunits in the cell.


A new pleiotropic bacteriophage P1 mutation, bof, affecting c1 repression activity, the expression of plasmid incompatibility and the expression of certain constitutive prophage genes.

In bacteriophage P1 an amber mutation in a new gene, bof, has been isolated. The bof-1 phage mutant exhibits a pleiotropic phenotype; bof product is non-essential, and acts as a positive modulator. In P1 bac-1 mutants, in which a dnaB analog product, ban, is expressed constitutively, the bof product activates ban expression both in the prophage state and in lytic growth: P1 bof bac prophages have a reduced ban activity and in lytic growth P1 bof bac phages show a lower ban activity than P1 wild type. This effect on ban activity is observed specifically in P1 bac-1 mutants; it is not mediated by the c1 repressor of the lytic functions (repressor of the ban operon) since this effect occurs even if the phage carries a heat sensitive c1 repressor. Thus we concluded that the bac mutation put the ban operon under an abnormal, unknown control, modulated by the bof product. P1 bof lysogens show an increased immunity to superinfecting P1 phage and are affected in their inducibility properties; in the presence of the altered c1-100 repressor, bof product is required for maintenance of lysogeny, as shown by the induction of P1 c1-100 bof-1 lysogens at 30 degrees. P1 bof superinfecting phage can be established together with a resident P1 bof prophage in a recA host, unlike P1 wild type which cannot form double lysogens. P1 bof double lysogens are unstable and segregate one or the other prophage. P1 Cm bof and P1 Km bof lysogens show higher levels of antibiotic resistance than the corresponding bof+ lysogens. The bof gene has been mapped, in an interval defined by P1 prophage deletion end points, far from both ban and c1. All bof phenotypes are reversed by single mutations.


Construction of an HpaI and HindII plasmid vector allowing direct selection of transformants harboring recombinant plasmids.

The construction of the vector plasmid PKN80 is described, which can be used as HpaI or HindII cloning vehicle with direct selection on transformants harboring hybrid plasmids. pKN80 carries the EcoRI.C fragment of phage Mu DNA coding for a killing function which is efficiently expressed upon transformation of pKN80 into Mu-sensitive bacteria. Cloning of DNA fragments at the single HpaI site of pKN80 results in insertional inactivation of the killing function. Whereas religated pKN80 molecules yielded only a few transformants, the transformation efficiency had been increased by a factor of at least ten when HpaI fagments of lambda DNA were added to the linearized vector prior to ligation. More than 90% of the transformants tested containted hybrid plasmids.


Failure of response of menopausal vasomotor symptoms to clonidine.

A double-blind crossover trial of clonidine failed to show any effect of the drug on menopausal vasomotor symptoms. A significant placebo effect was observed, the effect being greater in those with long-standing symptoms and a high neurotic index.


Picro-mercuro-formol: a fixative for histochemical studies.

The mixture of saturated solutions of pieric acid and mercuric chloride in the ratio of 9:9 forms a versatile fixative when added with 2 parts of 40% formaldehyde. It works satisfactorily with a number of histochemical reagents and even histological staining methods. The fixative may be applied for various types of tissue.


Quantitative image analysis: applications using sequential transformations.

The theoretical foundations of quantitative image analysis and its implementation in the Leitz T.A.S. have been discussed in the preceeding paper [4]. Especially the Mathematical Morphology and its recent developments have been pointed out. Sequential image transformations are a new approach of morphological analysis. Elementary transform steps, which are hardwired in the device, performed in specific sequences, which are implemented as macroinstructions in the programming language have proven to be a very useful tool in image analysis. A specific sequence of elementary steps will be referred to as morphological function. This new approach will be illustrated by four general examples: analysis of fibers, computing the number of edges for metallic grains, separation of overlapping cells, analysis of minerals which occur free and locked to another one.


A concept for stereological investigation of rat kidney.

An attempt is presented to adapt sampling procedures for stereological investigation of rat kidney. The proposed experimental design considers the zonal construction, in particular the structural and osmotic features of this organ. Every zone is treated as an own entity inregard to sample collection. In order to uncover possibly existing preferential orientation of structures the application of two mutually perpendicular section planes is recommended. Furthermore a method is described to determine absolute volumes and surface of structural components.


Isolation and partial characterization of exosporium from spores of a highly sporogenic mutant of Clostridium botulinum type A.

Homogeneous fragments of exosporium were isolated and purified from mature spores of a highly sporogenic mutant derived from Clostridium botulinum type A strain 190L. The exosporium was composed of three lamellae and showed a hexagonal array when negatively stained. The hexagonal array of isolated exosporium was resistant to sodium dodecyl sulfate, urea, dithiothreitol, and proteolytic enzymes such as trypsin, pronase, and nagarse, except for pepsin. The hexagonal array was partially disintegrated with 5 M guanidine-HCl and almost completely disrupted with 8 M urea in combination with 1% mercaptoethanol under alkaline conditions. The purified exosporium fraction was composed mainly of protein (69.1%) and lipids (13.8%). A small amount of amino sugars (2.5%) was present, but neutral sugars could not be detected. The exosporium protein had a predominantly acidic amino acid composition accompanied by low levels of cystine, methionine, and histidine.


Membrane glycoproteins shed in defence of the cells of the gastrointestinal tract.

The cells lining the gastrointestinal tract are exposed to various potentially harmful agents, including plant lectins and microorganisms. It is proposed that glycoproteins within the mucus layer of the gut protect against these agents. A hypothesis is presented that membrane glycoproteins are shed and, within the mucus close to the cell surface, block the binding of lectins or attachment of microorganisms to membrane receptors. A model is presented that demonstrates that such blocking can be achieved by soluble glycoproteins. The implications of this hypothesis in health and disease are discussed.


Efficacy of atenolol and oxprenolol in the treatment of arterial hypertension. A comparison.

Twenty-seven patients with mild to moderate arterial hypertension were treated "double-blind" with either atenolol of oxprenolol. Placebo was given for four weeks before the beta-blocking drugs were administered. Atenolol was given in doses of either 50 mg twice a day, 100 mg twice a day or 100 mg once a day for periods of four weeks. Oxprenolol was given in doses of either 80 mg or 160 mg twice a day for the same duration. Patients were assessed at the end of each four-week period. The mean blood pressure and pulse rate did not vary significantly for the two different dose regimens at which each drug was administered. There was no statistical difference between the reduction of systolic blood pressure produced by the two drugs, but there was a significant difference in the reduction in diastolic blood pressure in favour of atenolol (P less than 0.05 supine; P less than 0.01 erect). A single, 100-mg daily dose of atenolol was just as effective as 50 mg or 100 mg twice a day. Similarly, an 80 mg twice a day dose of oxprenolol was just as effective as that of 160 mg twice a day. Side effects for each drug were not statistically different from those recorded with placebo.


Labetalol and bendrofluazide: comparison of their antihypertensive effects.

A randomized double-blind crossover trial of labetalol, bendrofluazide and placebo was carried out in 18 patients with mild to moderate hypertension. The reduction in blood pressure observed with labetalol and bendrofluazide was similar. The plasma levels of labetalol showed a significant correlation with changes in blood pressure (r = 0.07; P less than 0.021); and plasma level determinations of this drug may be of value in the management of patients who receive antihypertensive agents.


Cimetidine in the treatment of reflux oesophagitis.

Twenty-two adult patients with active endoscopically proven reflux oesophagitis entered a double-blind trial of treatment with either cimetidine (1 g/day) or placebo. A repeat endoscopy was carried out at eight weeks after the beginning of treatment on 19 of the patients. The mucosa appeared macroscopically normal at eight weeks in 81.8% of the cimetidine-treated patients, while 90% of patients were described histologically as having a normal or unremarkable mucosa. The results in placebo-treated patients were 55.6% and 44.4% respectively. The cimetidine-treated patients consumed significantly less antacids than the placebo-treated patients, and the over all assessment of their well-being was significantly better (90.9% patient satisfaction with treatment in the cimetidine group compared to 18.2% in the placebo group). Two patients in the placebo group did not complete the trial due to continuing severity of their symptoms, while none in the cimetidine group felt that their condition had remained static or was worse.


[Carbohydrate-reduced diet and metabolism: about the influence of a 4-weeks isocaloric fat-rich, carbohydrate-reduced diet on body weight and metabolism (author's transl)].

The influence of a 4-weeks carbohydrate-reduced, fat-rich diet on 9 slightly overweighted men was investigated. Caloric and protein intake remained unchanged. 70% were fat, 20% carbohydrates. Glucose, cholesterol, triglycerids, serumproteins, urea, uric acid, sodium and potassium were measured in rest. At a 60 minutes bicycle ergometer test glucose, triglycerids, free fatty acids and glycerol were registrated before and after 20 and 60 minutes work. The intensity was 70% of the maximal oxygen uptake. These were the most important results: (1) Body weight decreased continuously. One of the causes is an elevated heat production. (2) Serum-protein and -glucose remained unchanged. Cholesterol and triglycerides were reduced. Serum-urea, -sodium and -potassium diminished continually. On the contrary, uric acid raised over the normal range. (3) Work performance was reduced for 20%. (4) Hypoglycemic values did not appear. The oxidation of fat by the working muscles, and fat mobilisation increased by a fat-rich diet.


Glucocorticoid hormone action: an overview.

An overview of glucocorticoid action is presented including aspects of historic, clinical, and physiologic interest. Despite the diversity of effects, glucocorticoid hormone action at the cellular level simplifies to a rather universal model that involves binding to a soluble receptor, followed by interaction of the complex with the chromatin and modification of gene expression. This unitary concept has important implications in pathology, pharmacology, and therapeutics. Finally, a definition of a glucocorticoid in terms of its receptor is presented, which we feel is better than the traditional terminology based on specific biological effects. It is hoped that this review will also provide the reader with a framework in which the various contributions in the Monograph can be integrated.


Factors influencing association of glucocorticoid receptor-steroid complexes with nuclei, chromatin, and DNA: interpretation of binding data.

Attempts to reconstruct, in a test tube, the steroid-hormone system of a responsive cell are fraught with enumerable difficulties. In this chapter I have attempted to point out some of the factors that affect receptor-steroid complexes and their interactions with acceptors. In most cases there is a quantitative influence of these factors on the level of steroid complex binding to acceptors. In some cases, selected experimental designs that neglect these factors and methods of presenting the observed data may lead to artifactual conclusions. Several of these problems should disappear when the prospect of pure receptor-steroid complexes [127, 147, 150, 181, 247, 248] becomes a common occurrence. Nevertheless much has already been learned about the interactions of complexes with acceptors, which in turn have been used to help formulate models of steroid-hormone action.


Glucocorticoid receptors and effects in human lymphoid and leukemic cells.

A variety of human leukopathic diseases including human acute lymphoblastic leukemia are responsive to glucocorticoids in a varying proportion of cases. We identified specific glucocorticoid receptors in human acute lymphoblastic leukemia cells. Their presence or absence was well correlated with both in vivo and in vitro responsiveness of these target cells to glucocorticoids. These data suggest that knowledge of glucocorticoid receptor status in human acute lymphoblastic leukemia may aid in selecting patients for therapy. Furthermore, these receptors exhibit significant quantitative differences in various subtypes of human leukemia, with null-cell lymphoblastic leukemia having approximately three times the mean number of receptors per cell as T-cell leukemias. These differences in receptor levels are associated with major differences in complete remission duration independent of other prognosticators of response such as patient age, white count, and cell surface markers. Specific receptors for glucocorticoids can also be identified in normal human peripheral blood monocyte fractions including unpurified peripheral blood lymphocytes, T, and non-T subcomponents of circulating lymphocytes and circulating monocytes. By criteria of quantity, of binding affinity, and specificity, these receptors appear to be similar to other classical glucocorticoid receptors. Receptors in human peripheral blood lymphocytes may be induced threefold on a per cell basis by treatment with the mitogen phytohemagglutinin. This is associated with a marked increase in glucocorticoid responsiveness.


Variations in cellular sensitivity to glucocorticoids: observations and mechanisms.

The spectrum of physiological, pathological, and genetic variations in sensitivity to glucocorticoids is reviewed. The receptor for these hormones is common to most mammalian tissues, and yet the responses are widely divergent. Although there may be differences in the receptors to account for some of this diversity, it is likely that it is largely due to cellular programming not involving the receptors. In addition to the intertissue differences in sensitivity, it is also clear that intra-tissue differences occur. The greatest amount of information has been accumulated with lymphoid cell systems and there are sensitivity differences to specific responses such as cell killing or effects on immunological functions. In these systems, there can be major variations in either the extent of the response (e. g., from mild growth inhibition to cellular killing) or whether any effect is observed. Further, dose requirements for certain responses can vary by several orders of magnitude. Within a given tissue there may be developmental changes in sensitivity that are not due to obvious changes in the receptor, and decreased sensitivity with aging that in some cases has been associated with changes in receptor binding activity. Finally, the cellular sensitivity can either be influenced by hormones and other factors that affect the ability of the glucocorticoid to elicit a particular response (in a synergistic or antagonistic manner), or the same function regulated by the glucocorticoid can be inducible by the steroid, appearing some time after administration of the steroid and disappearing after steroid removal. Genetic variations in sensitivity to glucocorticoids also occur. In humans these may be generalized, affecting glucocorticoid action in all responsive tissues, and could be important in the pathogenesis of certain diseases. Perhaps the most striking genetic alterations, however, are observed in cultured lymphoid and fibroblastic cells and in acute lymphoblastic leukaemia cells ordinarily growth inhibited or killed by the glucocorticoid. Mutant cell lines arise that are highly resistant and most of these have abnormalities in the glucocorticoid receptor. In some cases binding activity is totally lost, easily expalining the resistance. In other cases, there is a more modest reduction in binding or a change in receptor properties that give it increased or decreased nuclear and DNA binding activity. An analysis of these cell lines suggests that many of the defects are in some receptor property presently not understood that makes the receptor ineffective rather than the defect being due to the quantitative changes in receptor levels detected. The frequency of emergence of steroid-resistant cells can vary widely from about 10(-5) in S49 cells to less than 10(-8) in certain thymic cell lines...


Glucocorticoid receptors.

Glucocorticoid receptors are found in most mammalian tissues and have been studied in detail in a number of tissue culture systems. With cells that have not been exposed to steroids, the receptors are found in the cytoplasmic fraction from which they can be isolated and studied. Methods for studying glucocorticoid receptors depend on their high-affinity specific binding of radioactive steroids. The reversible interaction is intracellular. It follows Michaelian kinetics, at least in cell-free cytosol, and involves a thermodynamically homogeneous population of about 10 000 sites per cell. The receptor is an asymmetric, slightly acidic protein of about 100 000 daltons. It is very labile, especially in the unbound form. Binding activity depends on the integrity of thiol groups and perhaps on phosphorylation of amino acid residues. Although indirect, the evidence is overwhelmingly convincing that this protein is the physiologic glucocorticoid receptor. The time-kinetics of binding and dissociation are consistent with the sequence of events in glucocorticoid action. Various steroid analogs display binding characteristics predictable from their glucocorticoid activity. Loss of the binding protein from certain cultured cell lines is accompanied by unresponsiveness to glucocorticoids. The extensive tissue distribution of receptors parallels the extensive role of glucocorticoids in regulation. Finally, there is a strong correlation between nuclear binding of receptors and nuclear effects of the steroid. The glucocorticoid receptor can be distinguished from other glucocorticoid-binding proteins, based on their steroid specificity and physicochemical properties. There is no clear-cut demonstration that the receptor differs from tissue to tissue, and it is in fact very similar in various species. Unlike in other systems, receptor concentration does not seem to be regulated by its ligand or by other hormones. However, certain cases of hypo- as well as hypersensitivity to glucocorticoids appear to result from changes at the receptor level. The data indicate that the receptor can exist in inactive and active forms. The former predominate in the absence of steroid or when an angatonist is bound. Glucocorticoid agonists bind the active form, allowing it to be "activated" and subsequently bound to the nucleus. All of the receptors in isolated cytosol do not appear to be available for immediate occupancy by an agonist and this may be due to the time required for conversion of the receptors from inactive to active forms. The correlations between receptor binding and the glucocorticoid response indicate that the receptor is a rate-limiting factor in the magnitude and kinetics of the response, and this finding has important implications regarding mechanisms.


Regulation of gluconeogenesis by glucocorticoids.

1. Regulation of gluconeogenic substrate supply and modulation of the gluconeogenic pathway in the liver are both important in the control of gluconeogenesis by glucocorticoids. 2. Adrenal deficiency decreases the release of gluconeogenic and other amino acids from skeletal muscle during starvation. The effect is reversed by glucocorticoid replacement. The changes in amino acid release are accompanied by similar alterations in tissue amino acid levels and are not explained by alterations in net protein breakdown. Glucocorticoids do not alter protein catabolism and cause a small inhibition of protein synthesis. The biochemical alterations underlying the changes in amino acid metabolism induced by these steroids remain to be elucidated. Glucocorticoids may also regulate the supply of gluconeogenic substrates through permissive effects on the lipolytic action of catecholamines and other hormones in adipose tissue and on the glycogenolytic action of catecholamines on skeletal muscle. 3. Glucocorticoids are required for the increases in gluconeogenesis in starvation and diabetes. Part of their action is exerted directly on the liver and appears to involve modulation of P-enlopyruvate carboxykinase levels. Glucocorticoids increase the synthesis of this enzyme apparently through effects at the level of transcription. 4. Glucocorticoids exert permissive effects on the stimulation of gluconeogenesis in the liver by glucagon and epinephrine. The steroids are not required for cAMP generation or protein kinase activation by these hormones, but appear to act by maintaining the responsiveness of certain enzymes to the effects of the cAMP and alpha-adrenergic systems. It is proposed that this involves the maintenance of a normal intracellular ionic environment.


Glucocorticoids and the plasma membrane.

Glucocorticoids affect the composition and function of the plasma membrane in a variety of cell types. Cultured rat hepatoma (HTC) cells in tissue culture provide an excellent model system for analysis of such effects. In these cells, dexamethasone rapidly and dramatically inhibits the influx of amino acids sharing the A or alanine-preferring transport system. Inhibition is half-maximal within 2 h, and maximal after 6 h incubation with the hormone. The inhibition is rapidly reversed by insulin, and more slowly by removing the steroid. Microtubules and microfilaments are not apparently involved in this hormonal effect, but continuous protein synthesis is required for the glucocorticoid inhibition of transport. Dexamethasone also decreases the number of microvilli on the surface of HTC cells, increases their adhesiveness to a substratum, and dramatically decreases the production of plasminogen activator, but it does not affect the growth rate or plating efficiency of the cells. Variant cell lines stably resistant to dexamethasone inhibition of plasminogen activator production have been isolated using an agar-fibrin overlay technique to detect protease production by individual colonies of HTC cells. The hormonal resistance to inhibition of protease production is associated witha  maintenance of inducibility of other glucocorticoid-regulated functions and therefore is not apparently secondary to abnormal or absent glucocorticoid receptor, but due to a lesion in a later step in hormone action specific for plasminogen activator. Combined genetic and biochemical analysis of such dexamethasone-resistant variants should facilitate study of the hormonal regulation of specific membrane phenotypes and of the role of proteases in this regulation.


Glucocorticoids and lysosomes.

Steroids in general and glucocorticoids in particular affect lysosomes in various ways. The explanation of these effects remains in dispute, however. Theories include the view that steroids interact directly with lysosomal membranes, that steroids provoke induced changes in lysosomes, and that classical steroid receptors originate in lysosomes. Experiments pertaining to these views are discussed, particularly with respect to steroid specificity and tissue specificity of effects and to dose-response considerations.


Structure and conformation of glucocorticoids.

The approaches currently available for determining the structure and conformation of glucocorticoids are reviewed. We discuss the optimization of steroid geometry based on the relative molecular energy calculated by a Westheimer equation. This method permits an extensive description of steroid molecules in a state free of external constraints and which can be assumed to correspond to the minimum internal energy. The structures, conformations, surface areas, and volumes of fifteen steroid molecules that interact with the glucocorticoid receptor have been studied. The basic structure of the A ring is a 1 alpha,2 beta-half-chair, whatever the substitutions. Rings B and C are semi-rigid chairs virtually uninfluenced by substituent groups. In contrast, the shape of the D ring depends on the nature and environment of the substituents. As to the fundamental conformation of the side chain, the steroids fall into two categories, depending on the pressure of a 17-hydroxyl group. For a given molecule, the energy changes associated with conformations of the side chain other than that corresponding to the minimum energy have also been explored. The hypothesis is formulated that receptor binding requires a particular conformation of the side chain. Finally, the overall shape of the molecule can be influenced by the summation of minor but numerous changes brought about by various substitutions, such as 11 beta-hydroxyl, which increases the convexity of the molecule. These investigations should help in elucidating structure-activity relationships for glucocorticoids. They may improve our knowledge of the interaction between these hormones and their receptor and of the molecular mechanism of glucocorticoid action.


[Structural changes in clinical pediatrics with special reference to small departments (author's transl)].

Changing facts and developments in clinical pediatrics in Germany during the past years have been analysed and discussed. Facts which altered the quantity and quality of clinical pediatrics are demonstrated by own observations, and informations collected from all pediatric departments in the FRG and West-Berlin. The need for sufficient numbers of staff to meet with the changing and intensified tasks of clinical pediatrics is emphasized.


Mutagenic effects of benzo[a]pyrene after metabolic activation by hepatic 9000 g supernatants or intact hepatocytes.

The activating capacities of isolated rat hepatocytes and 9000 g supernatant from these cells with respect to the mutagenic effect of benzo[a]pyrene on Salmonella typhimurium TA100 were investigated. No mutagenicity of benzo[a]pyrene was found with the cell-mediated assay, unless the hepatocytes were disrupted after pre-incubation with benzo[a]pyrene or the intracellular glutathione content was reduced. It is suggested that a retention of active metabolites and an effective detoxication may account for the absence of mutagenic response.


Effect of X-irradiation on frameshift and missense mutations in Saccharomyces cerevisiae.

In cell populations of Saccharomyces cerevisiae homogeneous for sensitivity to X-irradiation, induction of base insertions/deletions and base substitutions was quantitatively analyzed in a reversion system. The repair mechanisms phenotypically unexpressed in the sensitive cell fraction and fully operating in resistant cells did not affect point mutations of either type.


Antagonism by propidium of petite induction by ethidium and ethidium azide in Saccharomyces cerevisiae.

Propidium, a phenanthridinium dye similar to ethidium, did not induce petite mutations in non-growing yeast cells in contrast to ethidium. Combined exposure to ethidium and an excess of propidium for periods up to 2 h resulted in the expected petite induction expressed after subsequent plating on growth medium. As incubation was continued with propidium, the numbers of petites declined on subsequent plating whether the drug had been added before, during, or after the mutagenic treatment by ethidium. Propidium decreased petite induction by the monoazide analog of ethidium when applied before but not after photolytic attachment of the drug.


A recA-dependent mutator of Escherichia coli K12: method of isolation and initial characterization.

A number of mutator strains of E. coli were isolated using histochemical techniques which allow the identification of a single mutator colony on agar plates with as many as 2000 colonies. Several mutators isolated in this way were found by P1-mediated transduction to map to the proA--proB region of the E. coli chromosome. The map position of these mutators is very close to that of the conditional mutator, mutD. However, in contrast to mutD, one of these newly isolated mutators was suppressed in a thermosensitive recA strain at 43 degrees C, but not at 30 degrees C. This mutator mutation has been named mut-8. Besides being dependent upon recA, mut-8 is also dependent upon growth in enriched medium for the expression of its mutator activity. The mutator activity of mut-8 was found to be recessive to the wild-type allele.


The mutagenicity of nitrosamides in Salmonella typhimurium.

34 nitrosamides (10 nitrosoalkylcarbamates, 2 nitrosoalkylnitroguanidines, 12 nitrosoalkylureas, 6 substituted nitrosoalkylureas, and 4 cyclic nitrosoalkylureas) were tested for mutagenicity in Salmonella. All were direct-acting mutagens of varying potency.


In vivo conversion of sodium azide to a stable mutagenic metabolite in Salmonella typhimurium.

Salmonella typhimurium TA1530 and G46 strains growing in minimal medium supplemented with sodium azide produce a stable mutagenic metabolite which is not azide. The production of this metabolite is restricted to the log phase of bacteria grown in the presence of azide. The metabolite is highly mutagenic in DNA-repair defective base-substitution strains TA1530 and TA1535, but ineffective in frameshift strains TA1538 and TA1537. The metabolite induces mutations in resting cells of the TA1530 strain.


Mutagenic cholesterol preparations.

Naturally air-aged commercial samples of USP or reagent-grade cholesterol contain components which are mutagenic towards Salmonella typhimurium strains TA1537, TA1538 and TA98. These mutagenic components are associated with the polar cholesterol autoxidation products, but identity of the mutagenic components has not been achieved. Pure crystalline nonmutagenic cholestrol free from autoxidation products becomes mutagenic towards these strains upon heating at 70 degrees in air or following exposure to 60 Co gamma-radiation.


Mutagenicity evaluation of the two antimalarial agents chloroquine and mefloquine, using a bacterial fluctuation test.

The two antimalarial agents chloroquine and mefloquine have been tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1537 and TA1538. Chloroquine was found to revert strain TA1537 at concentrations of 100 and 250 micrograms/ml, most likely due to intercalation. No mutagenicity was found with mefloquine at concentrations up to 2.5 micrograms/ml, neither without nor with metabolic activation by Ca2+-precipitated rat liver microsomes. Higher concentrations of mefloquine and chloroquine inactivated the bacteria.


Genetic toxicity of procarbazine in bacteria and yeast.

Procarbazine [N-isopropyl-alpha-(2-methylhydrazino)-p-toluamide hydrochloride] is used to treat Hodgkin's disease. This compound was tested in vitro without and with S10 fraction from mice liver (microsomal assay) using Saccharomyces cerevisiae strain D7, Salmonella typhimurium (strains TA98, TA100, TA1535) and in vivo in Swiss albino mice (host-mediated assay) using D7. Procarbazine, without S10 fraction, is highly toxic and induced mitotic crossover, gene conversion, and reverse mutation in D7. It had a toxic effect on all the Salmonella strains; but did not induce reverse mutations at the histidine loci. Procarbazine, with S10 fraction, was less toxic and did not induce genetic effects in yeast or Salmonella. In the host-mediated assay, no genetic effects were seen.


Genetic effects of hydralazine.

Hydralazine and its acetone condensation product (ACP) were found to induce base-pair substitution mutations in the Salmonella/microsomal activation test system and to display genetic toxicity in the PolA+/A- test system. Incubation with a rat-liver microsomal fraction did not affect the genetic toxicity of either compound. Other derivatives of hydralazine, including the major metabolite, 3-hydroxy-methyl-s-triazolo-[3,4a]phthalazine, did not yield any evidence of genetic toxicity nor were they metabolically convertible to a toxic product. Therefore, individuals who convert hydralazine to MTP slowly, the "slow acetylators", would be expected to be at risk.


Dominant lethal assay of some hair-dye components in random-bred male rats.

Male rats were exposed to maximally tolerated doses of 5 hair-dye components in a dominant lethal test. Each component was tested at 3 dosage levels with 15 random-bred male rats per level. The highest dose, selected on the basis of subacute toxicity testing, generally reduced weight gains without being lethal. Freshly prepared solutions were injected i.p. at 1 ml/kg 3 times a week for 10 weeks. Rats injected with dimethylsulfoxide and triethylenemelamine served as solvent and positive controls, respectively. A majority of rats survived the treatment at the levels tested and were mated to two virgin females each per week for 2 weeks. The females were sacrificed at midterm of pregnancy and examined for live and dead implants. Dominant lethality was evaluated on the basis of 4 criteria: dead implants per pregnant female, dead implants per total implants, proportion of females with one or more dead implants, and proportion of females with two or more dead implants. 2-Nitro-p-phenylenediamine, 2,4-diaminoanisole sulfate and 2,5-diaminoanisole sulfate produced negative responses, whereas m-phenylenediamine and 4-nitro-o-phenylenediamine induced weak dominant lethality in the first trial. On retesting these weakly positive components, both m-phenylenediamine and 4-nitro-o-phenylenediamine produced negative responses.


Mutagenic activity of thiram in Ames tester strains of Salmonella typhimurium.

The mutagenic activity of thiram was investigated in 4 histidine-requiring strains of Salmonella typhimurium (TA1535, TA100, TA1538, TA98) with and without activation by liver microsomes. In strains TA1535 and TA100, thiram induces mutations without metabolic activation. The presence of rat-liver microsome fraction, cysteine or glutathione abolish its mutagenic activity in these strains. In contrast, thiram requires metabolic activation for the expression of its mutagenic activity in TA1538 and TA98 strains. The compounds containing the sulphydryl group abolish mutagenic activity of thiram in these strains, too.


Opportunistic lung infection due to "Pittsburgh Pneumonia Agent".

Eight immunosuppressed patients had pneumonia due to Pittsburgh Pneumonia Agent (PPA), a gram-negative, weakly acid-fast bacterium cultivatable only in embryonated eggs and guinea pigs and distinct from Legionella pneumophila. The diagnosis was established by isolation of the agent from lung or visualization of the organism in lung tissue. The clinical presentations, radiographic abnormalities and pathology were not specific. The most consistent feature associated with the disease was the recent institution of daily high-dose corticosteriod therapy in all patients. Five of the eight patients died despite broad-spectrum antibiotic and antituberculous therapy. Anti-microbial activity against PPA was demonstrated for sulfamethoxazole combined with trimethoprim, for rifampin and for erythromycin with an egg-protection assay. Serologic studies with an indirect fluorescent-antibody technic suggested that seroconversion or high titers may be a sensitive test for PPA disease. PPA appears to be a newly recognized cause of life-threatening bacterial pneumonia in immunosupressed patients.


Opportunistic pneumonia: a clinicopathological study of five cases caused by an unidentified acid-fast bacterium.

Five patients had opportunistic pulmonary infection caused by acid-fast bacilli, unusual clinical presentations and a unique pathological picture. Clinically, these cases mimicked septic pulmonary emboli or bacterial pneumonia. The infection was temporally related to high-dose corticosteroid therapy, given for renal-transplant rejection in four patients and for therapy of lymphocytic lymphoma in one. Histologic sections of lung-biopsy or autopsy material showed an acute suppurative pneumonia with dense alveolar infiltration by neutrophils, without granuloma formation or caseous necrosis. Predominantly intracellular acid-fast bacilli were present. The organism failed to grow in culture on routine bacterial, fungal and mycobacterial mediums. This unusual and possibly new acid-fast organism is a probable cause of suppurative pneumonia in impaired hosts receiving corticosteroid therapy.


Failure of antiplatelet and anticoagulant therapy to improve patency of grafts after coronary-artery bypass: a controlled, randomized study.

Fifty patients who underwent aortocoronary saphenous-vein bypass-graft surgery were randomly assigned to one of three groups to determine the effects of antiplatelet or anticoagulant therapy on graft patency. Twenty-four patients served as controls; 13 patients received aspirin (325 mg three times a day) and dipyridamole (75 mg three times a day); and 13 patients received closely regulated warfarin therapy. Medications were begun on the third post-operative day. Six months after surgery, all patients underwent coronary angiography to assess graft patency. There were no statistically significant differences between groups in various clinical, hemodynamic and angios, 27 of 33 grafts (82 per cent) with aspirin and dipyridamole and 29 of 37 grafts (78 per cent) with warfarin (P less than 0.5), all patients had at least one patent graft. Postoperative treatment either with aspirin and dipyridamole or with warfarin failed to improve the patency of the grafts.


Cyclic oscillations of basal plasma glucose and insulin concentrations in human beings.

In a study of whether oscillations in plasma glucose and insulin occur in human beings, plasma samples were taken at one-minute intervals from 10 normal subjects for periods lasting between one and two hours. In five subjects the basal plasma insulin concentrations cycled regularly, with a mean period of 13 minutes and mean amplitude of 1.6 mU per liter (11.5 pmol per liter). A concurrent plasma glucose cycle was demonstrated, with a mean amplitude (after averaging to minimize random error) of 0.05 mmol per liter (1 mg per decliter). The average plasma glucose cycle was two minutes in advance of the plasma insulin. In the subjects with less regular plasma insulin cycles, a similar plasma glucose rise was demonstrated two minutes before the insulin rise. These phase relations are compatible with the presence of a negative-feedback loop between the liver and pancreatic beta cells that regulates both basal plasma insulin and glucose concentrations, although the cyclic beta-cell secretion could be independent of plasma glucose.


Cimetidine for anastomotic ulcers after partial gastrectomy. A randomized controlled trial.

In a randomized double-blind multicenter trial, 15 outpatients with endoscopically proved anastomotic ulceration after Billroth I or Billroth II partial gastrectomy received cimetidine, 1 g daily over eight weeks, or a placebo. All patients also received antiacid. The ulcer healed completely in all seven cimetidine-treated patients and in one of the eight placebo-treated patients (P less than 0.01). Ulcers not healed during the double-blind phase of the trial were all subsequently healed on open cimetidine treatment. There was a trend toward improvement of daytime symptoms in favor of cimetidine (P = 0.06), and nighttime symptoms were significantly relieved during the initial four weeks of cimetidine treatment P = 0.02). We conclude that cimetidine, 1 g daily, promotes healing of anastomotic ulcers after partial gastrectomy.


Effect of amphotericin B on the metabolism of Aspergillus fumigatus.

Action of amphotericin B on the growth and metabolism of Aspergillus fumigatus has been investigated. The fungus proved to be very sensitive to amphotericin B, showing complete inhibition of growth at 0.5 units/ml. Amphotericin B suppressed the exogenous and endogenous respiration and glycolysis of A. fumigatus as well as the assimilation of various glycolysis and TCA cycle intermediates. Addition of cations and cholesterol failed to reverse the action of amphotericin B. The treated mycelium released a variety of cellular constituents and it is inferred that the antibiotic effects the permeability of A. fumigatus cells. In experiments with 32P labelled mycelium phosphorus compounds leached out in concentrations which were dependent on the antibiotic dose, period of contact, incubation temperature and metabolic state of the fungus.


Cell-mediated immune responses in mice infected with Fonsecaea pedrosoi.

Time course of cellular and humoral immune responses in mice infected with Fonsecaea pedrosoi was investigated by using an antigen prepared from culture filtrate of this fungus. Mice were infected by intravenous injection with yeast-like cells of the fungus. Viable fungus was recovered from the brain of the infected mice until the 36th day after inoculation, and from the other organs examined until 14th to 16th day. Inflammatory lesions were observed in the brain, lung, heart, liver, spleen, kidney and intestine during the first 30 days after inoculation. Macrophage migration inhibition factor response in these mice was insignificant until 8 days after inoculation. A significant response was developed at day 10 and persisted until day 63. This response returned negative by 95 days after inoculation. Lymphocyte transformation response of these mice was negative until 4 days after inoculation. At day 6 blastogenic index increased to 1.5, and at day 10, 14 and 16 the indices were 1.8, 2.4 and 1.7 respectively. Precipitin response to this fungus could not be detected in these mice until 16 days after inoculation, Positive results were obtained at day 21 and lasted until 51 days after inoculation. The precipitin titers, however, did not exceed one fold in any of these mice.


Population changes induced in Candida albicans by nalidixic acid.

Cells of Candida albicans plated on media containing nalidixic acid (Nal) either die, adapt physiologically to Nal-tolerance or mutate ot Nal-resistance. The fraction of a population exhibiting each response depends on the growth phase of cells when plated and their nitrogen and carbon nutrition and growth temperatures before and after plating. Nal induces Nal-resistant mutants in very high frequency but only at 37 C on plates containing i) glucose as primary carbon source and ii) adenine, a sulfur amino acid or a representative of the glutamic acid family of amino acids. Nal does not affect either forward mutation to caffeine-resistance or reverse mutation from histidine auxotrophy to prototrophy. Nal-resistant mutants produce minute colonies on N-ALfree medium, respire oxidatively and are unusually sensitive to inhibitors of oxidative phosphorylation. They revert spontaneously to wild type at very high rates but can be propagated indefinitely in the absence of Nal by serial selection and replating of minute colonies. Cellular inactivation and induction of Nal-resistant mutants are greatly affected by specific inhibitors of mitochondrial macromolecular syntheses. The presence of chloramphenicol or erythromycin during exposure to Nal prevents cell death and mutation but has no effect on adaptation to Nal-tolerance. Growth on acriflavin or ethidium bromide enhances resistance of cells to inactivation when subsequently plated on Nal containing media. It is concluded that Nal-induced cellular inactivation and mutation to Nal-resistance, but not adaptation to Nal-tolerance, result from damages to the mitochondrion which are fixed or promoted by macromolecular syntheses within the mitochondrion. Implications of these findings for the therapeutic use of Nal are discussed.


Directive segregation in the basis of colE1 plasmid incompatibility.

Incompatibility between colE1 plasmids in Escherichia coli can be explained by competition for a limited number of replication sites. These sites ensure directive segregation of plasmids to daughter cells on cell division. The number of sites can be more than one only if a linear segregation mechanism is postulated.


Direct expression in Escherichia coli of a DNA sequence coding for human growth hormone.

DNA coding for human growth hormone was constructed by using chemically synthesised DNA in conjunction with enzymatically prepared cDNA. This 'hybrid' gene was expressed in Escherichia coli under the control of the lac promoter. A polypeptide was produced having the size and immunological properties characteristic of mature human growth hormone.


Captopril (SQ 14225) depresses drinking and aldosterone in rats lacking vasopressin.

Angiotensin II is dipsogenic, and vasopressin (ADH) regulates renal water excretion. Together, these hormones govern overall mammalian water balance. The Brattleboro rat with inherited diabetes insipidus (DI) lacks ADH and is therefore a convenient model with which to elucidate mechanisms regulating water metabolism. In the present studies, angiotensin II has also been removed from DI rats by the administration of an inhibitor (captopril, SQ 14225; D-2-methyl-3-mercaptopropanoyl-L-proline) of the enzyme which converts angiotensin I, the relatively inert component of the renin-angiotensin system, to angiotensin II, the biologically active substance. SQ 14225 reduced the drinking rates, and after 6 days lowered peripheral plasma aldosterone concentrations were associated with hyperkalaemia. We conclude that the polydipsia of diabetes insipidus partly results from elevated plasma renin activities and angiotensin II concentrations seen in this syndrome. Further, the apparent hypoaldosteronism of DI Brattleboro rats reflects differences in both tissue usage of the steroid and adrenocortical sensitivities associated with polyuria, hyperosmolarity and possibly potassium wasting.


The importance of Ca2+-mediated phospholipase A2 activation for stimulus-evoked PGE2-release from rabbit splenic capsular strips.

Rabbit splenic capsular strips released PGE2 when contracted by noradrenaline. Contraction as well as PGE2-release were Ca2+-dependent. In the presence of Ca2+ the ionophore A 23187 induced a long lasting contraction and a vigorous PGE2-release. In Ca2+-free medium the ionophore was ineffective. Introduction of Ca2+ to splenic strips preincubated with the ionophore in the absence of Ca2+ reinstalled contracture and PGE2-release. The PGE2-release induced induced by the PG-precursor arachidonic acid was Ca2+-independent. Addition of phospholipase A to splenic strips elicited a strong PGE2-release. These results indicate, that the first step of stimulus evoked PG-release is the Ca2+-mediated activation of phospholipase A2. This enzyme liberates from membrane bound phospholipids C-20-fatty acids, which then serve as substrates for the PG-synthetizing enzymes. Therefore the rate limiting step in stimulus-evoked PG-release seems to be rather the phospholipase A2 activity than the state of activation of one of the PG-synthetizing enzymes.


Membranous nephropathy: a radioimmunologic search for anti-renal tubular epithelial antibodies and circulating immune complexes.

In an effort to elucidate immunopathogenic of membranous nephropathy (MN), freshly collected sera from patients with biopsy proven MN were assayed of circulating immune complexes (ICs) by the Raji cell method and for anti-renal tubular epithelial (RTE) antibodies by a newly established radioimmunoassay (RIA) and by indirect immunofluorescence. 6 of 26 MN patients tested by the Raji cell assay had detectable circulating ICs. However, 5 of these 6 patients had other medical conditions which might also explain the IC reactivity. 29 MN patients and 11 patients with other glomerular diseases had no demonstrable circulating anti-RTE antibodies. This study suggests that if RTE antigens possess a nephritogenic potential for man it is probably only rarely expressed. The inconstant detection of circulating immune complexes in idiopathic MN raises an speculation as to their immunopathogenic significance.


A detailed analysis of the 1-hour posttransplant renal biopsy. Its value in predicting future outcome of kidney transplants.

A detailed histological assessment of the 1-hour posttransplant biopsy has been made in 65 cadaver and related renal transplants. There was a lack of correlation between the numbers of polymorphonoclear leucocytes per glomerulus, the prescence and severity of thrombi, vascular changes, interstitial scars, the presence of absence of immune deposits, and the renal function of the patients at 90, 180 and 365 days. The only positive findings was a statistically significant correlation between the glomerular tamponade at 90 days and the renal function. This was not borne out at 180 and 365 days.


Skin muscle biopsy in patients with various nephropathies.

Skin-muscle and kidney biopsy specimens were obtained from 43 patients with glomerulonephritis and from 5 normal subjects. 14 of the 43 patients had mesangial IgA nephropathy and in 12 the immunofluorescence study of skin-muscle biopsy showed IgA deposits. Ultrastructural research performed on 4 of these patients showed some electron-dense deposits on the walls of the muscular capillaries or in the perivascular connective tissue which could represent the immune aggregates. The remaining 29 patients had nonsystemic chronic glomerulonephritis with various immune deposits; only in 7 of these patients was the immunofluorescence study positive. The said study suggests that immunofluorescence examination of skin-muscle biopsy specimens could be diagnostically important.


Renal 'salt wasting' despite apparently normal renal, adrenal and central nervous system function.

The inability of the kidney to conserve sodium appropriately in response to a restricted sodium intake is reported in a subject who had entirely normal renal and adrenal function and no evidence of central nervous system disease. Subsequent transplantation of his left kidney to his son afforded a unique opportunity to assess renal sodium conservation in each kidney separately: both the patient and his son were unable to achieve balance in response to dietary sodium restriction after transplantation, indicating that the sodium wasting was due to a process intrinsic to both kidneys. The assessment of intrarenal hemodynamics with xenon-133 washout prior to nephrectomy provided insight into the role of intrarenal hemodynamics in the maintenance of sodium homeostasis in man. The xenon washout curve failed to disclose an identifiable second most rapid exponential (CII). This finding, in concert with previous studies demonstrating that diuretic agents with a primary action in the ascending limb of the loop of Henle caused a marked slowing of CII flow, raises the possibility that limited perfusion in the outer medulla of the kidney may be rate-limiting for sodium conservation in man.


Assessment of Brescia-Cimino fistula blood flow rates in pediatric patients.

Blood flow rates were measured using an isotopic dilution method in the main drainage segments of Brescia-Cimino arteriovenous fistulas in 9 pediatric patients who had undergone successful renal transplantation, 3 patients on hemodialysis and 2 patients with end-stage renal disease. Blood flow rates varied from 68 to 882 ml/min. The possibility that such high flow rates may have deleterious effects on limb growth, cosmetic appearance and cardiac function without any benefit to the patient is discussed.


Steroid-cyclophosphamide pretreatment of kidney allograft donors. A control study.

34 cadaveric donor grafts were randomized in a blind study of the effect of pretreatment of 5 g each of methylprednisolone and cyclophosphamide on kidney graft outcome. There was no difference in overall survival or functioning after 3, 6 or 12 months between grafts from pretreated (33 kidneys) or control (29 kidneys) cadaveric donors. In addition, this pretreatment protocol did not modify the recipient immune response against B-lymphocyte alloantigens which developed in unsuccessful transplants. Our data, thus, neither confirm the high rate of kidney graft survival attributed to cadaveric donor treatment nor the supposition that treatment is effective in suppressing recipient antidonor B-lymphocyte antibodies.


Dipropylacetate and hyperglycinemia.

Hyperglycinemia was induced by dipropylacetate (DPA) (dose 14-41 mg/kg per day) in 10 Finnish epileptic patients with neurologically disabling diseases of various kinds. When comparing the levels of glycine in plasma, cerebrospinal fluid and urine from the patients with 10 otherwise comparable patients, and normal values there was a fourfold excretion of glycine in urine. All the treated patients had elevated CSF (49 +/- 23 mumol/l) and plasma (475 +/- 76 mumol/l) glycine concentrations. The plasma/CSF glycine ratio was not influenced by the treatment. In comparison with corresponding values in nonketotic hyperglycinemia and ketotic hyperglycinemia, this condition probably arises in a manner similar to secondary hyperglycinemia in organic aciduria. DPA probably influences metabolism in a way similar to some metabolites of amino acids in organic acidurias.


[Markers of type B viral hepatitis].

The discovery of Australian antigen (HBsAg) has led to an increasing deal of knowledges about the virus of type B hepatitis (HBV); several markers of HBV have been detected and are becoming disposable for clinical and epidemiological purposes. The HBsAg is carried by 3 types of particulate structures discovered by electron microscopy as small spherical particles having diameter around 22 nm, long filaments and spherical particles having an overall diameter of the 42 nm (Dane-particle) with an electron-dense core. Dane-particle core contains circular double-stranded DNA molecules and an enzyme, the DNA polymerase. At present, Dane-particle is thought to represent the HBV, having properties consistent with those of a complete virus. Four antigen/antibody systems related to viral type B hepatitis have been discovered; they have been designated with the following nomenclature: HBsAg/anti-HBs, HBcAg/anti-HBc, HBeAg/anti-HBe, epilon antigen/anti epilson. The availability of the HBV markers for clinical purposes will permit a better understanding of the sequence of the biological reactions as well as of the clinical and epidemiological features concerning this viral infection: incubation period, acute disease, resolution, chronic carrier state, actively or passively immunized subject, persistent or subsided infectivity, prognosis.


[Electrophoretic mobility of lymphocytes in multiple myeloma and bronchial cancer].

The electrophoretic mobility of peripheral lymphocytes was studied in normal subjects and patients with myeloma and bronchial carcinoma. Distinct differences in anodic mobility were noted between the sub-populations in all three groups. Total lymphocyte migration was significantly slower in the two tumour groups.


[The hyperaminoacidurias with special reference to cystinuria].

Increased urinary excretion of free amino acids is a sign which requires determination of the underlying cause. Physiological aspects of tubular transport with emphasis on the role played by transport proteins or carriers are briefly discussed. Subsequently we present a resumptive classification. In prerenal hyperaminoaciduria the urinary excretion of amino acids just reflects the error in amino acid metabolism. Depending on tubular reabsorption of the amino acid involved in the metabolic error three types can be distinguished: the overflow, the competitive and the non-threshold hyperaminoacidurias. Renal hyperaminoaciduria can either be specific for individual or group-related amino acids, or generalized involving all amino acids. With the exception of classical cystinuria the various hyperaminoacidurias have more theoretical than clinical importance. Generalized hyperaminoaciduria is always a symptom of severe tubular disturbance which can be produced by various metabolic diseases, intoxications or deficiency states. Finally we present our experience in the treatment of cystinuria with mercaptopropionyl-glycine.


A new simple technique for the identification of antibodies against double stranded DNA: preliminary communication.

A new, simplified assay for detecting double stranded DNA antibodies by indirect immunofluorescence using fixed human metaphase chromosomes as substrate is described. The technique has the advantage of ease of preparation, clarity of reading, specificity for double stranded DNA and apparently greater sensitivity than immunofluorescent assays currently available. Thirty of 92 systemic lupus erythematosus sera were found to have double stranded DNA antibodies using the metaphase preparation, compared with 23 positive sera detected with the Crithidia luciliae technique. No double stranded DNA antibodies were detected in 90 sera from patients with rheumatoid arthritis. The human metaphase chromosome assay could replace existing techniques for identification of double stranded DNA antibodies.


Captopril and propranolol in mild and moderate essential hypertension: preliminary report.

A randomised open trial was carried out to test the efficacy and safety of Captopril and propranolol in 20 patients with mild and moderate essential hypertension. Mild and moderate was defined as a resting supine diastolic pressure of 100 to 120mmHg at the end of a placebo period. Patients were then randomised into Captopril (12) or propranolol (8) group. After a four-week dose ranging period, they were maintained on the treatment drug, or, if still uncontrolled, the other drug was added. The patients showed good overall blood pressure response to both treatment regimes. Three patients in the Captopril group developed a rash. The significance of the effectiveness in essential hypertension of a drug which acts to block the renin-angiotensin-aldosterone system is discussed.


Radiocolloids in the treatment of ovarian cancer.

Though radiocolloids have been used for over twenty years in the treatment of ovarian cancer, no comprehensive review of the literature exists. This review incorporates the historical development of radiocolloids, discusses the physical properties and physics of the two commonly used colloids, radioactive gold (Au-198) and radioactive phosphorus (P-32), discusses instillation and distribution of intraperitoneally administered P-32, and reviews result of treatment. The conclusions reached from this review are that Au-198 is a very different radiocolloid than P-32; that the dose of clinically used P-32 is determined from an empirically determined dose of Au-198; and that the dose of 100 mCi to 150 mCi of Au-198 yields significantly greater radiation than 10 mCi to 15 mCi of P-32. The complications associated with Au-198 are results of a possibly excessive dose, with a significant gamma component, delivered over a shorter period of time. It is clear that in greater than 99% of distintegrations each beta particle is associated with a gamma component. The review of the literature also shows that no propsective randomized well controlled study has indicated the effectiveness of P-32 over other treatment modalities. The clinical applicability of P-32 awaits prospective randomized trials and the ability to accurately determine dose and dose-distribution in-vivo.


Control of perinatal infection by traditional preventive measures.

A 9-year review of nosocomial infections occurring in the department of obstetrics and gynecology of a major teaching hospital demonstrated the dramatic effect of strict asepsis and isolation on postoperative infection rates. With rigidly enforced preventive measures and without the use of prophylactic antibiotics, the rates of wound infection and endometritis and the relative prevalence of gram-negative aerobic organisms cultured from these infections could be reduced substantially. The role of anerobes in these infections was not established, since adequate techniques for the accurate diagnosis of anaerobic infection were not available at the time of this study. The relative importance of the numerous infection-control measures used is unclear. It is apparent, however, that by implementing all traditional methods of asepsis, virtually all types of nosocomial infection could be controlled, and the increased prevalence of gram-negative infections observed during the past decades could be reversed.


Trophoblast growth in the lungs of mice.

Trophoblast giant cells grow from ectoplacental cones of 7 1/2 days' gestation that are implanted in the lung from the surface, intrabronchially, or via the circulation. When blastocysts of 3 1/2 days' gestation are implanted in lung, their behavior varies according to the route of implantation and the ages of the recipient mice. Blastocysts produced trophoblast giant cells when implanted in the bronchial tree. Trophoblast giant cells were not seen in adult lungs when blastocysts were introduced via the circulation. However, they were seen in 12-day-old mice injected in the same way 10 days previously. Something is apparently liberated from adult lung vessels which interacts with the blastocyst or zona pellucida and causes rapid degeneration of trophoblast. Experiments designed to extract this agent are in progress in the hope that the agent may be useful in the treatment of human trophoblastic disease.


Spontaneous feminization and menstrual function developing during puberty in Turner's syndrome.

An 18-year-old patient with Turner's syndrome presented with cyclical vaginal bleeding and spontaneous development of secondary sexual characteristics. She demonstrated classic features of Turner's phenotype, and a culture of blood lymphocytes revealed a 45,XO karyotype. The patient's plasma and urinary estrogen concentrations were similar to those in normal adult women in the late proliferative phase. In contrast, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels were elevated to values seen in postmenopausal women. Dynamic testing revealed no estrogen response to human menopausal gonadotropin (/MG) and a paradoxical fall in estrogen after administration of human chorionic gonadotropin (hCG). Administration of luteinizing hormone-releasing hormone (LH-RH) resulted in an exaggerated response of both gonadotropins. Dilatation and curettage revealed endometrial hyperplasia, and a laparotomy revealed the presence of 2 gonadlike structures. The histologic diagnosis was lutein cyst. Karyotyping of lymphocytes, skin from the abdominal incision, and tissue from both gonadal structures revealed a 45,XO karyotype without evidence of mosaicism.


Mechanism of single-stitch failure of tubal ligation: a morphologic appraisal.

The widespread demand for a simple, safe, and reversible sterilization procedure has attracted the attention of clinicians and researchers. A simple ligation with a single suture of nonabsorbable linen, involving a very small segment of tube (1 mm) in the isthmic portion about 1 cm from the isthmic ampullary junction, was used in 463 multiparous women. Of the 363 cases followed for a period of 1--3 years, there have been 52 failures. In 10 unsuccessful cases total abdominal bilateral salpingectomy was performed, and a detailed histologic study of these 20 tubes was instituted in an attempt to understand the mechanics of failure of the single-stitch technique. This is the first such study to be reported.


[Ultrastructure of the spermatozoids of the pulmonate molluscs, Trichia hispida and succinea putris].

The ultrastructural study of spermatozoa of hermaphroditic land molluscs Trichia hispida and Succinea putris has shown that the general pattern of their structure is similar. At the same time differences were found in the fine structure of acrosome, centriolar region (neck) and mitochondrial derivate (accessory nucleus). The mitochondrial derivate of spermatozoa of both the species contains three secondary spirals of varying length (vs. one-two in other species of pulmonate molluscs).


[Ligandin localization in the gonadal cells of rats at various stages of ontogeny].

Ligandin, a protein binding some carcinogens, steroids and other substances in the rat liver, has been found by means of indirect immunofluorescence in the gonad cells of different types: embryonic and mature Leidig cells in testes, ovarian thecal cells at the maturation stages (theca interna, atretic follicles and interstitial cells) and luteal cells of corpus luteum at pregnancy. Ligandin is found, thus, in cells which belong to various lines of cell differentiation. The functional role of ligandin is discussed.


[Influence of the age of the recipient on the manifestation of the effect of allogeneic inhibition].

The expression of allogenic inhibition was studied when transplanting 10(5) cells of bone marrow of C57BL mice to the lethally irradiated recipients (CBA X C57BL) F1 of different age (2 to 11 months). In the control experiments the bone marrow cells at the same dose were introduced to the lethally irradiated syngenic (C57BL) mice. The most pronounced inhibition of the parental stem cells proliferation was registered in 2 months old recipients F1 (4.7 times), it was somewhat weakened in 3 months old and animal in 4--11 months old recipients (1.1 to 1.8 times). The thymectomy of adult recipients F1 did not eliminate the expression of allogenic inhibition.


Modified Wheeler orbicularis overlap procedure for senile entropion.

Senile entropion is a condition involving the lower lid due to aging. Surgical correction has been directed to either increasing the vertical or horizontal tension on the tarsus. While all are good, each has its shortcomings with resultant recurrences. The procedure we are presenting is a combination of features which has given us the least recurrence rate. These include horizontal shortening of skin and orbicularis muscle and positioning of orbicularis straps so as to prevent their riding up on the tarsus. Figures illustrating the procedure in detail as well as pre and postoperative photos are shown.


Repair of total and subtotal symblepharons.

Large adhesions between a globe and an eyelid reform after surgical sectioning. Techniques to prevent reattachment have relied on mucosal grafts or plastic shields to prevent opposing raw surfaces from touching each other. A procedure is described using neither graft nor plastic shells which enables the traumatized surfaces to be opposed to uninjured mucosa until healing has been completed. This procedure makes use of techniques which are usually employed to repair other kinds of ophthalmic problems: after the symblepharon is dissected from the cornea, it is used to form the inner surface of the lid; Beta radiation is applied and bare sclera is not covered; the globe is then rotated with traction sutures and tethered to the lateral canthus until spontaneous epithelialization is achieved.


Mechanical ectropion repair using tarsal rotation sutures.

Chronic mechanical ectropion may persist despite elimination of its causal factors. Four such cases are presented in which traditional surgical management was not appropriate. Instead, tarsal rotation sutures were applied, eliminating the ectropion and deepening the inferior fornix in each case. This technique is presented in detail, along with a discussion of its mechanical principle which establishes the infraorbital rim as the center of rotation of the eyelid, anchoring the sutures and placing an inward torque on the tarsal plate.


Epidemiology, microbiology and management of otitis media.

Epidemiologic studies indicate that the maximum incidence of otitis media (OM) occurs 6 to 24 months of age. Features associated significantly with recurrent OM include sex (males greater than females), race (white greater than black) and having a sibling with recurrent OM. Persistence of middle ear effusion (MEE) following acute OM is frequent; 40% of children have MEE one month after onset of OM. Streptococcus pneumoniae and Haemophilus influenzae are the most frequent organisms isolated from MEE of children with acute OM. Viruses and mycoplasma are uncommonly present in MEE. Therapy of acute OM includes antibiotics active against the two major organisms. Appropriate management of recurrent episodes of OM or persistent MEE is unclear but current studies include use of prophylactic antimicrobial agents and a multitype pneumococcal vaccine.


Current advances in perinatal medicine.

The enhancement of pulmonary maturation with the resultant ability to prevent hyaline membrane disease has given rise to the use of a number of drugs experimentally including corticosteroids, thyroxine, aminophylline, heroin, and several suggested others for this purpose. In clinical use to date only the corticosteroids have been widely utilized in man but it is clear that these agents are capable of effecting an advancement in the maturation of the surfactant system of the lung and a subsequent reduction in the incidence of severity of hyaline membrane disease. However, all agents which act by the enhancement of maturation also carry with them a resultant arrest to replication of tissues and there have been demonstrable effects on both the lung and possibly the brain of the organisms to whom these agents have been administreted. In this connection it is, therefore, important to realize that the advantages gained from these agents may be counterbalanced by disadvantages from their usage and that a reasoned and careful approach in each individual case is mandatory when use of such agents is considered. Experimental and clinical studies suggest the possibility that Vitamin E acting as a free radical scavenger may be important in the prevention of oxygen toxicity both to the eye (retrolental fibroplasia) as well as ameliorating the oxygen component of the destructive effects of respirator lung disease (bronchopulmonary dysplasia). The action of Vitamin E under these circumstances is not dependent upon its actual quantitative level in the plasma of premature infants but in utilization to its excess. Although phototherapy has been universally and widely used since 1958, precise studies of its mechanism of action have not been revealing. Recent work has suggested that following exposure to photoirradiation, bilirubin in its unconjugated form can be seen to appear in the excretory bile ducts suggesting the transport of unconjugated bilirubin through the liver. More recent evidence has also suggested that this occurrence is the result of internal rotation following light exposure of one of the double bonded rings which effectively converts the molecule from a lipid soluble to a water soluble and thereby excretable form. These studies would not only account for an appropriate explanation of the quantitative reduction in bilirubin observed but carry with them the other important consideration that the amount of light exposure necessary to accomplish this is strikingly less than the previously considered photooxidation reaction. The clinical implications of these findings may suggest a reevaluation of the quanta of light which has been used for purposes of phototherapy to date.


Asymptomatic bacteriuria in adolescent girls: II. Screening methods.

Of 500 asymptomatic adolescent girls who were screened for bacteriuria by three methods-dipslide (Uricult), dipstrip (Microstix-3 reagent strips), and home nitrite test (Microstix-Nitrite reagent strips)-eight cases (1.6%) were detected: 6/8 by dipslide and dipstrip; 5/8 by nitrite testing. The false-positive-rate (greater than 10(4) colonies/ml) of the dipslide test was 6.4%, and the dipstrip test, 2.8%. A history of vaginal discharge was not associated with "contaminated" specimens. False-positive nitrite tests were reported by 0.6% of the patients who returned the postcards. Overall, 70.4% of the patients returned the postcards for the home nitrite test. The patients were divided by method of payment (Medicaid vs non-Medicaid) in order to provide an approximation of socioeconomic status; non-Medicaid patients were significantly more likely to return postcards than Medicaid patients (75.8% vs 63.7%). Of the group reporting previous urinary tract infection, 79% of both Medicaid and non-Medicaid patients returned postcards, suggesting that a prior experience with the diagnosis increased compliance with a home test.


Modern management of tetanus neonatorum.

Five infants with neonatal tetanus were managed with neuromuscular blockade and positive pressure ventilation. All survived. Tetanus was frequently confused with neonatal seizures at time of presentation. All infants were delivered at home. Mean age of onset of symptoms of poor suck, hypertonicity, and generalized spasms was 5.4 days with hospital admission at 6.4 days. Mean duration of mechanical ventilation was 34 days. Although hospital management of tetanus neonatorum may be expected to have a satisfactory outcome, it represents a costly and complex approach to a preventable disease.


[Successes of modern malariology].

Malariological situation analysis in the USSR and world over testifies to increasing malaria incidence particularly in the countries where earlier it was almost fully eradicated. Malaria epidemics in these countries (posteradication epidemics) is a new phenomenon in malariology. Temporary success of the global campaign of malaria eradication has led to a deadlock in malariology which during the past 10 years is being successfully overcome. Achievements of modern malariology are reviewed in the article on three levels of the host-parasite relationships: a cell, organism and population. Particular significance of the research in malaria vaccines is emphasized.


Impaired cell-mediated immune response in patients with congenital rubella: correlation with gestational age at time of infection.

Lymphocyte transformation, interferon, and leukocyte migration inhibition factor synthesis were studied in purified lymphocyte cultures for 20 children with congenital rubella and 18 healthy children (seven susceptible and 11 immune to rubella). Lymphocyte transformation after phytophemagglutinin stimulation was significantly lower in children with congenital rubella as compared to healthy controls. Responses to purified rubella virus were absent in the susceptible controls and absent or at least two times lower in congenital rubella children than in immune controls. After purified rubella virus stimulation, leukocyte migration inhibition factor production was detected in all immune controls, but in none of the susceptible controls,  or the congenital rubella-infected children. The results varied with gestational age of intrauterine infection: the impairment of cellular immune response, both after phytohemagglutinin or rubella virus stimulation, was more severe in the children infected in the first two months than in the latter stages of gestation.


Countercurrent immunoelectrophoresis in group B streptococcal disease.

Countercurrent immunoelectrophoresis was used for the detection of group- and type-specific antigens in the body fluids of 61 infants from St. Louis and Indiana with group B streptococcal infections. Urine concentrated using an Amicon filter yielded the highest percentage of positive results; 81% were positive in the St Louis group. When three body fluids (urine, CSF, and blood) were available, at least one was positive for group B streptococcus in 95% of the cases. This study demonstrates the applicability of this test in a tertiary care facility (St Louis) and in smaller hospitals (Indiana) with access to central laboratory.


Comparison of urinary lactic dehydrogenase with antibody-coated bacteria in the urine sediment as means of localizing the site of urinary tract infection.

Two noninvasive methods of localizing the site of urinary tract infection, urinary lactic dehydrogenase (LDH) isoenzymes and antibody coating of bacteria in the urinary sediment, have been prospectively compared with the bladder washout technique in a series of children with urinary tract infection. Fifteen children had infection localized in the upper tract. Urinary LDH isoenzymes correctly localized the infection in 14 children; however, the infection was correctly localized by the antibody coating of bacteria in only eight patients (P less than .02). Fourteen children had lower tract infection by the bladder washout technique. Urinary LDH isoenzymes localized the infection in all 14 children, whereas the antibody coating correctly localized the infection in ten children (P less than .05). This study shows the urinary LDH isoenzyme pattern to be a more accurate technique than the detection of antibody-coated bacteria for localizing the site of urinary tract infection.


Nephritis in children and young adults with systemic lupus erythematosus and normal urinary sediment.

One male and seven female patients (aged 6 to 26 years) with systemic lupus erythematosus (SLE), normal urinalyses, and normal biochemical tests of renal function, had renal biopsies to determine if significant nephropathy existed. Several had active SLE in other body systems at the time, either clinically or as evidenced by low serum complement and high native DNA antibody levels. The renal biopsy specimens were studied by light, fluorescent antibody, and electron microscopy. Three patients had a generalized segmental, two had a focal segmental, and one had a generalized diffuse proliferative glomerulonephritis. In addition, one patient had minimal glomerular findings with interstitial inflammation. All eight patients were found to have moderate immune complex deposition by immunofluorescence and/or electron microscopy studies. The absence of clinical renal involvement in patients with SLE does not preclude ongoing active and "silent" glomerular damage with moderately severe proliferative changes.


The occurrence of salmonella in waste water from Danish slaughterhouses. A quantitative study.

In waste water from 11 slaughterhouses, the occurrence of salmonellas were followed continually during a whole week. Thirty-eight samples out of 66 were contaminated. The mean concentrations of salmonellas varied between 0.9 and 668 per 100 ml. Only 4 different serotypes were found viz., S. agona, S. indiana, S. senftenberg, and S. typhimurium. This distribution of serotypes is incompatible with epidemiological data from humans, animals, and feed-stuffs. It is suggested that certain serotypes of Salmonella may become established in sewage systems of slaughterhouses or that propagation of salmonellas may take place in the sewer rats. In one particular plant, large numbers of S. senftenberg were found during the whole period of sampling. It is pointed out that discharge of such heavily polluted waste water into water courses may constitute a public health hazard if the recipient is used for irrigation of vegetables or pastures.


[Bacterial infections in renal homotransplant recipients. Results of regular bacteriological controls (author's transl)].

Bacterial infections were studied in 51 immunosuppression kidney transplant recipients. Eighty infectious episodes occured in the first six post-transplantation months. Forty percent of these episodes were nonfebrile, detected during the routine bacteriological follow-up examination. Out of these 80 bacterial infections, 59 were localized (47 in the urinary tract, nine in the surgical wound and three in the lungs) and 21 had spread. Neutopenia occured in 13 patients, in association with septicaemia in eight of them. Usually septicaemia appeared before neutropenia. In 12 cases, infectious episodes (10 septicaemias and two localized infections) and transplant failure were closely related. The bacterial strains responsible for these infections were essentially: Staphylococcus spp (39), Streptococcus spp (14), Klebsiella pneumoniae (9), Escherichia coli (9). Most of the localized infections cleared up when treated with a specific antibiotic. Seven failures of antibiotic treatment were observed in 21 cases of generalized infections. The importance of regular post-transplantation bacteriological check-up in detecting and treating infectious complications in graft recipients must be emphasized.


[Drug-induced active hepatitis (author's transl)].

Drug-induced active hepatitis is rare. The three main drugs incriminated are oxyphenisatine, alphamethyldopa and isoniazid. Despite the histological appearance suggestive of chronic active hepatitis, such forms of hepatitis, and in particular those due to alphamethyldopa and isonizid, follow an acute rather than a chronic course. The course is usually rapidly favourable when the responsible drug is stopped, provided the histological lesions are not those of multilobar necrosis or cirrhosis.


Measurements of paralytic shellfish poisons. A review of biological and chemical procedures.

Paralytic shellfish poisons (PSP) are derivatives of a tetrahydropurine base, which includes saxitoxin, 1-hydroxy saxitoxin, and 11-hydroxy saxitoxin sulfate. These compounds are produced by Gonyaulax catenella and other dinoflagellate species, and the algal toxins (phycotoxins) contaminate shellfish during the filter-feeding process performed by the bivalve molluscs. Increased incidence of algal blooms and outbreaks of PSP food poisonings have been encountered in recent years, resulting in a demand for closely monitoring of shellfish for these toxins. The existing biological and chemical procedures for analysis are reviewed. The mouse bioassay is commonly used in routine analysis, but is nonspecific. Several of the chemical procedures are specific and more sensitive than the mouse bioassay. The disadvantage, however, seems to be that not all known PSP compounds can be quantitiatively measured by any single chemical procedure.


[Tubo-uterin implantation: microsurgical technic (author's transl)].

Fourteen tubo-uterin implantations for endometriosis of the horn and infertility have been carried out. They have been made with microsugery with a reversed transcornual T access. The setting of a retractor pushing off the edges of the uterin muscle allows the removing of the pathological zone, allows also not to hurt the vascularisation during the dissection of the nodule, and allows also to suture edge to edge the tube to the uterus under microsurgical control. This original process allows a new setting up of the continuity (Ten permeabilities cheked with hysterography 2 pregnancies and 2 obstructions among 14 cases after a passing of less than 1 year).


[Shock. A review (author's transl)].

Shock is defined as a secondary condition constituting a complication to a primary disease of which more than 100 are recorded in the literature. Shock is characterized by prolonged circulatory inadequacy leading to insufficient tissue perfusion and cell death. According to etiology shock is classified into three main groups: hypovolemic, vasogenic and cardiogenic shock. Taking hypovolemic shock as a model the pathgenesis of shock is presented. Hypovolemia acts on the baroreceptors giving rise to a sympatho-adrenal response resulting in increased vasoconstriction, which again leads to viscerocutaneous ischemia. This phase is known as the ischemic anoxic or centralized shock phase. Without treatment this phase develops into the second socalled stagnant anoxic or paralytic shock phase. "Irreversible shock" is discussed. The pathogenesis of vasogenic and cardiogenic shock is mentioned and compared with hypovolemic shock. It is emphasized that the sympatho-adrenal response is the central and common feature in every shock development. Special reference is made to septic shock with its outstanding circulatory conditions (arteriovenous shunting). Lacticacidemia and metabolic acidosis are described as the most important metabolic alterations in shock. With reference to pathenesis the main clinical symptoms of shock are presented: increased heart rate, initially pale later hyperemic, congested and terminally cyanotic mucosae, increased capillary filling time, cold skin and low body temperature. All these signs are related to the sympatho-adrenal response. It is pointed out that the patient in shock is depressed. Inevitably the primary disease will modify the shock symptoms. Hyperemia with edema, hemorrhages and thrombosis in organs and tissues are morphological manifestations of shock. Later microscopically detectable degenerative and necrotic alterations develop, and there are signs of intravascular coagulation (hyaline thrombi and spheres). Due to the rather nonspecific macroscopic alterations a post mortem shock diagnosis necessitates for completion histology and/or a clinical shock diagnosis. Some of the most important shock-provoking primary diseases dealt with in veterinary practice are mentioned along with their possible shock pathogenesis. Referring to the shock pathogenesis the therapy is discussed. The first and indispensable therapeutical measure in treating shock per se is increasing the circulating blood volume, Balanced electrolyte solutions are preferred. Examples of composition, doses (up to 80--200 ml/kg body weight) and infusion rate (initially 15--30 ml/kg body weight during the first 10--20 min., then quantum satis) are given.


The effect of protein-binding on the excretion of three sulphonamide preparations in the milk of dairy cows, examined by chemical and microbiological methods.

The relationship between the protein-binding of sulphonamides and their excretion in the milk of dairy cows was studied using three preparations commercially available in Finland. After a preparations containing sulfadiazine and sulfadimidine was given intravenously to dairy cows the drugs were excreted into milk to a greater extent than in the case of sulfamethoxypyridazine and especially of sulfaphenazole given similarly. An inverse relationship was found between the degree of protein-binding in the serum and the excretion into milk. The antimicrobially active concentrations of sulphonamides in serum and milk persisted for less than 24 hours when the doses recommended by the manufacturers were used. From a pharmacological point of view the sulfadiazine-sulfadimidine combination seems to be the drug of choice. Although no traces of sulphonamides were detected 48 hours after the dosing, the question of milk residues could not be answered because the minimum detection level of the methods used in the study was approximately 1 microgram/ml. The IDF standard method for the detection of penicillin in milk is not suitable for the detection of sulphonamide residues in milk.


[The localisation of urinary tract infections using the tetrazolium nitroblue test (author's transl)].

In order to study the value of the tetrazolium nitroblue test (TNB) in the localisation of urinary tract infections, the authors performed this test in 33 patients with a urinary tract infection and in 20 control children suffering from no bacterial infection. The 33 infected children were divided into three groups on the basis of clinical and bacteriological criteria (dysuria, frequency, fever, abdominal pain, inflammatory syndrome, immunofluorescence of bacteriuria, serum antibody levels): -- group A (8 children): upper urinary infection -- group B (14 children): lower urinary infection -- group C (11 children): urinary infection of undetermined site. The number and percentage of TNB positive polynuclears was significantly higher in the children of group A than in the children of group B (p 0.001). By contrast there was no significant difference between the results obtained: in the non-infected control group and in group B -- in group A and group C. These results, confirming a previous study by Bjorksten and de Chateau, show the value of the TNB test in the localisation of urinary tract infection in the child.


[Aorto-coronary venous bypass. The value of microsurgical techniques (author's transl)].

The application of microsurgical techniques to myocardial revascularisation surgery by aorto-coronary venous bypass makes it possible to extend the indications for operation to patients with severe coronary lesions, generally considered to be unsuitable. A technique of vascular suture in two layers combined with a widening angioplasty is described. Their value is illustrated by the blood flow studies and the clinical results in 66 bypass performed on 32 patients.


[Systemic lupus erythematosus and anti-native desoxyribonucleic acid antibodies: clinical and laboratory comparison of the results given by two methods of detection (author's transl)].

Simultaneous application of two methods for the detection of anti-native DNA antibodies (indirect immunofluorescence on DNA fibres and Farr's radioimmunological method with native DNA labelled C14) to 53 serum samples from 32 patients suffering from systemic lupus erythematosus (SLE) indicated a certain number of discordances between the results obtained. Correlations between clinical and laboratory findings were found, such that certains types of anti-native DNA antibodies (n) may be specific to certain clinical manifestations of SLE: a good correlation was seen between She fixation of anti-C14 nDNA and the titre of IF anti-nDNA in the group of SLE with renal involvement, but was not found in the group of SLE with central nervous system involvement. There was no parallel between the course of renal involvement and anti-nDNA antibody titres, without treatment being directly responsible. The significance of the presence or absence of anti-DNA antibodies with the two methods of detection used is not of any single significance. Several types of anti-nDNA antibodies are present in a given specimen of lupus serum. Certain anti-nDNA antibodies are not detected or are at the limit of a positive result in the presence of neurological manifestations. From a practical standpoint, the use of several laboratory tests is desirable for the possible investigation of SLE.


[Artificial pancreas: clinical and therapeutic value in 11 patients (author's transl)].

Fluctuations in blood glucose levels which occur in the brittle diabetic explain the difficulty of blood glucose control by insulin therapy as used at the present time. The artificial endocrine pancreas is a bedside device capable of maintaining blood sugar value within a normal range by the administration of adequate amounts of insulin or glucose given at the optimal time. The glycemic control of 11 brittle diabetics is improved during the 5 days after a 24 hour connection with the AEP. A better determination of adequate subcutaneous doses of insulin, based on the circadian insulin profile infused by the artificial endocrine pancreas, achieved the improvement of glycemia. Glucagon profile preceeding, during and after AEP is also discussed. Using a method of two injections daily of a mixture acterapid and semi-lente, as a general rule it is found that approximately fifty per cent of the dose must be given in the acterapid form. Glucagon value decreases during the 24 h artificial endocrine pancreas. This is suggestive of a possible role of hyperglucagonemia in brittle diabetes.


Rapid print-readout technique for sequencing of RNA's containing modified nucleotides.

A rapid, simple, and highly sensitive method for sequence analysis of RNA was developed, which consists of the following steps: (i) controlled hydrolysis of the RNA by brief heating in water; (ii) (32P)-labeling of 5'-hydroxyl groups of the fragments produced in (i); (iii) resolution of labeled fragments by size on polyacrylamide gels giving the familiar "ladder"; (iv) contact transfer ("print") of the ladder from the gel to a PEI-cellulose thin layer; (v) in situ treatment of the ladder with RNase T2 resulting in the release of 5'-(32P)-labeled nucleoside-3',5' diphosphates; (vi) contact transfer and thin-layer separation of (32P)-labeled nucleotides on PEI-cellulose in ammonium sulfate and ammonium formate solvents; (vii) autoradiography. The chromatographic behavior of the 4 major and 18 modified nucleotides was determined. The positions of major and modified nucleotides in the sequence can be read directly from the separation patterns displayed on X-ray film. As this is the only sequencing method presently available that allows one to display and identify directly the positions in the RNA chain of major and modified nucleotides, no additional procedures are required to analyze the latter.


Nucleotide sequence of starfish initiator tRNA.

The nucleotide sequence of starfish ovary initiator tRNA was determined to be pA-G-C-A-G-A-G-U-m1G-m2G-C-G-C-A-G-U-G-G-A-A-G-C-G-U-G-C-U-G-G-G-C-C-C-A-U-t6A-A-C-C-C-A-G-A-G-m7G-D-m5C-C-G-A-G-G-A-psi-C-G-m1A-A-A-C-C-U-C-G-C-U-C-U-G-C-U-A-C-C-AOH. The sequence was determined by a combination of the two different post-labeling techniques. Two-dimensional cellulose thin-layer chromatography was adopted for analysis of 5'-terminal nucleotides of tRNA fragments produced by formamide treatment. The nucleotide sequence of starfish initiator tRNA is very similar to that of mammalian cytoplasmic initiator tRNAs, but has seven different nucleotide residues and two modifications: residue 55 is psi instead of U, and residue 26 is unmodified G instead of m2G.


Binding of Escherichia coli ribosomal proteins to 23S RNA under reconstitution conditions for the 50S subunit.

The RNA binding capacity of 50S proteins from E. coli ribosomes has been tested under improved conditions; purified proteins active in reconstitution assays were used, and the binding was studied under the conditions of the total reconstitution procedure for the 50S subunit. The results are: 1) Interaction of 23S RNA was found with 17 proteins, namely L1, L2, L3, L4, L7/L12, L9, L10, L11, L15, L16, L17, L18, L20, L22, L23, L24 and L29. 2) The proteins L1, L2, L3, L4, L9, L23 and L24 bound to 23S RNA at a level of about one copy per RNA molecule, whereas L20 could bind more than one copy (no saturation was observed at 1.8 copies per 23S RNA), and the other proteins bound 0.2--0.6 copies per RNA. 3) L1, L3, L7/L12 showed a slight binding to 16S RNA, L26 (identical with S20) strong binding to 16S RNA. 4) The binding of L2, L7/L12, L10, L11, L15, L16 and L18 was preparation sensitive, i.e. the binding ability changed notably from preparation to preparation. 5) All proteins bound equally well to 23S RNA in presence of 4 and 20 mM Mg2+, respectively, except L2, L3, L4, L7/L12, L9, L10, L15, L16 and L18, which bound less strongly at 20 mM than at 4 mM Mg2+.


Isolation of a single polypeptide leucyl-tRNA synthetase from bakers' yeast.

A single polypeptide of leucyl-tRNA synthetase (LRS) has been purified from budding bakers' yeast by a modification of the procedure published earlier. On denaturing polyacrylamide gel electrophoresis LRS was one band corresponding to molecular weight of 120,000 +/- 5,000 daltons. Variable amounts of LRS with a similar molecular weight but which dissociated into equal subunits of 58,000 were also isolated. The affinities (KM) for substrates for this form of the enzyme were similar to those previously reported for the dimeric form of the enzyme.


Release of 7-methylguanine residues whose imidazole rings have been opened from damaged DNA by a DNA glycosylase from Escherichia coli.

Double-stranded DNA containing 7-methylguanine residues whose imidazole rings have been opened, i.e. 2,6-diamino-4-hydroxy-5-N-methylformamido-pyrimidine residues, may be prepared by treatment of DNA with dimethyl sulfate followed by prolonged incubation at pH 11.4. These substituted formamidopyrimidine residues are actively removed from DNA by a DNA glycosylase present in E. coli cell extracts. The enz;me shows no apparent cofactor requirement and has a molecular weight of about 30 000. The release of ring-opened 7-methyl-guanine residues is due to a previously unrecognized activity, different from the three known E. coli DNA glycosylases that release uracil, 3-methyladenine, and hypoxanthine from DNA. This enzyme may serve to repair a major secondary alkylation product in DNA. In addition, it may remove nonmethylated purines, whose imidazole rings have been opened, from X-irradiated DNA.