Plasma dehydroepiandrosterone in pregnant and nonpregnant women.

From 157 plasma samples taken randomly throughout normal pregnancy and from 42 plasma samples of nonpregnant women, total plasma dehydroepiandrosterone was measured by a method using Amberlite XAD-2 column chromatography at 45degreesC, enzyme hydrolysis, radioactive internal standard, thin-layer chromatography and gas-liquid chromatography after trimethylsilyl ether derivative formation. The following values for dehydroepiandrosterone were obtained: from individual, nonpregnant samples, (n = 25) 69.6 +/- 10.6 mug/100 ml (S.E.M.); from the pool of nonpregnant samples (n = 17) 67.7 mug/100 ml; from individual samples, 6-12 weeks of gestation (n = 32) 48.5 +/- 5.7 mug/100 ml (S.E.M.); from individual samples, 13-18 weeks of gestation (n = 13) 45.9 +/- 7.7 mug/100 ml (S.E.M.); from individual samples, 19-24 weeks of gestation (n = 20) 42.9 +/- 6.9 mug/100 ml (S.E.M.); from individual samples, 25-30 weeks of gestation (n = 22) 41.7 +/- 6.8 mug/100 ml (S.E.M.); from individual samples, 31-36 weeks of gestation (n = 31) 39.5 +/- 6.1 mug/100 ml (S.E.M.); from individual samples, 37-43 weeks of gestation (n = 29) 37.6 +/- 3.6 mug/100 ml (S.E.M.); and from the pool sample, 37-43 weeks of gestation (n = 10) 25.4 mug/100 ml. This study demonstrates a significant decrease of total plasma dehydroepiandrosterone throughout the course of normal pregnancy in individual and pooled plasma samples, thus confirming previous reports. These plasma hormone changes are discussed in relation to production and utilization of this steroid in pregnancy.


Intra-erythrocytary enzymes before and after splenectomy.

The activity of the intraerythrocytary enzymes glucose-6-phosphate dehydrogenase, pyruvate kinase, glutathione reductase and ATPase was measured before and after splenectomy in 13 patients with congenital hemolytic anemia and 3 patients suffering from chronic thrombocytopenia. All patients were treated successfully, as reflected by clinical and basal hematological parameters. Glucose-6-phosphate dehydrogenase and pyruvate kinase were significantly depressed after splenectomy. It was not possible to set up prognostic criteria of splenectomy from the intraerythrocytary enzymes.


The role of low affinity antibody in immune complex disease. The quantity of anti-DNA antibodies in NZB/W F1 hybrid mice.

The level and avidity of anti-DNA antibody in the serum of New Zealand Black/White (NZB/W F1) hybrid mice has been determined. The results show that there is an age and sex-related variation in the avidity of this antibody. In mice of both sexes, the avidity of circulating anti-DNA antibody increases up to 5 months of age; thereafter the avidity falls with increasing age. These variations are more marked in males, but female mice consistently have lower avidity anti-DNA antibody than males. Thus the time of onset, time course and severity of the murine lupus syndrome in NZB/W F1 mice are associated with the presence of increasing levels of low avidity anti-DNA antibody in the serum. These results are discussed in the context of the possible role of low avidity antibody in immune complex disease.


In vitro reactivity of lymphocytes obtained from uraemic patients maintained by heamodialysis.

Lymphocytes obtained from uraemic patients maintained on intermittent haemodialysis had a normal ability to respond to and stimulate allogeneic lymphocytes obtained from normal subjects in the mixed lymphocyte culture (MLC) reaction. The response of these uraemic lymphocytes to PHA and pokeweed mitogen (PWN) was also normal. Uraemic plasma from eight out of twenty-six patients studied, however, possessed blocking factor activity which suppressed the MLC reactivity of normal random donors and also the mitogenic response of allogeneic lymphocytes but not of autologous uraemic lymphocytes. The blocking factor activity was attributed to a non-dialysable factor present in the plasma of the patients investigated.


Drug-induced haemolytic anemia.

Drug administration causes 16 to 18 per cent of cases of acquired immune haemolytic anaemia. The pathogenesis of erythrocyte sensitisation by drug-related antibody with or without fixation of complement is variable and there is a relationship between the responsible drug, the mechanism of red cell sensitisation, clinical manifestations, and laboratory methods of diagnosis. Drugs such as phenacetin and quinidine form a complex with the antidrug antibody, and the immune complex attaches to red cells usually fixing complement and causing acute intravascular haemolysis. Other drugs (e.g. penicillins) when given in high doses coat normal red cells in vivo and some patients develop a high titre IgG antidrug antibody which reacts with the coated cells. Haemolytic anaemia may develop, with red cell destruction being primarily extravascular. Cephalosporins cause positive direct antiglobulin tests in a small percentage of patients either by the same mechanism as penicillins or by modification of the red cell membrane leading to non-immunological absorption of serum proteins. Haemolytic anaemia has been reported only rarely. A few drugs (notably alpha-methyldopa) cause the development of autoimmune haemolytic anaemia. A knowledge of clinical manifestations and laboratory aids to diagnosis are necessary to distinguish immunohaematological abnormalities caused by drugs fron other causes. Drugs may also cause haemolytic anaemia by nonimmunologic mechanisms as a result of oxidative denaturation of haemoglobin. Factors which make red cells particularly susceptible to the effects of oxidant drugs are intraerythrocytic metabolic abnormalities or the presence of unstable haemoglobins.


Stimulation of chondroitin sulfate proteoglycan production by chondrocytes in monolayer.

Chondrocytes in monolayer undergo morphological and biochemical changes which culminate in the establishment of cartilage nodules in vitro. Chondroitin sulfate or heparin, added to the culture media of these cells, stimulates the production of chondroitin sulfate proteoglycan over the entire period of culture with a maximum effect during the log phase of growth. In addition, a lag of 2-3 hours is required before an increase in sulfate incorporation into polysaccharide is observed. The responsiveness of chondrocytes is influenced by several factors, such as cell density, conditioned media and enzyme treatment. Furthermore, puromycin abolishes the endogenous as well as the stimulated synthesis, demonstrating the necessity for core protein synthesis in both synthetic processes. Addition of beta-D-xylosides (which presumably act as initiators of chondroitin sulfate polysaccharide synthesis) and chondroitin sulfate, concurrently, stimulate sulfate incorporation to levels higher than either agent alone, indicating that these compounds act by different mechanisms.


The linkage region in the polypeptide of pig costal cartilage proteoglycan.

Proteoglycan from pig costal cartilage and fragments obtained by proteolytic digestion were characterized by equilibrium ultracentrifugation and amino acid analysis. The proteoglycan extractable in 4 M guanidinium chloride yielded, after proteolytic digestion with trypsin and chymotrypsin, a chondroitin sulfate peptide containing four chains of polysaccharide. The unextractable residue yielded chondroitin sulfate peptide containing only two chains. The amino acid composition indicated a fairly uniform spacing between all four chains with an average of eight amino acid residues between the serine residues involved in linkage. Following the alkaline sulfite elimination-addition reaction, free peptide was isolated and found to contain one unsubstituted serine residue for every two linked glycosidically. Glycine and glutamic acid were the only two amino acids sufficiently abundant to be part of an invariant sequence near to serine residues destined to be glycosylated. The linkage region of the polypeptide also contains some substituted serine residues which do not carry a full chondroitin sulfate chain.


Studies on the mechanism of hyaluronate lyase action.

Hyaluronate lyase from streptococci group A was purified by chromatography on DEAE-Sephadex A-50 and on Biogel P-150, thereby enriching it about 1,000-fold and separating it into two enzyme fractions with the same amino acid composition. The photooxidation of hyaluronate lyase in the presence of methylene blue results in rapid inactivation of the enzyme. The histidine content of the enzyme is decreased considerably, but also the content of methionine, tyrosine, and lysine is lowered. The enzyme is inhibited, but incompletely so, by N-tosyl-L-phenyl-alanine-chloromethyl ketone (TPCK) and N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK). Hyaluronic acid methyl ester, prepared form hyaluronic acid and diazomethane, is not split by hyaluronate lyase (EC 4.2.2.1.). Hyaluronic acid methyl ester is not a competitive inhibitor of hyaluronate lyase. For the mechanism of the enzymatic elimination reaction a proton transfer between histidine of the enzyme and the carboxylate group of hyaluronate is proposed.


Content and composition of glycosaminoglycans in human knee joint cartilage. Variation with site and age in adults.

The content and composition of the matrix of human articular cartilage from two sites - the condylar summits and the upper, posterior surfaces of both lateral and medial sides - of the lower femoral epiphysis of 12 autopsy cases (age 18-46 years) were analyzed. A significantly higher content of both chondroitin sulphate and especially of keratan sulphate was found in cartilage of the condylar summits. The solubility profile of keratan sulphate showed changes indicating a difference between the metabolism of cartilage of the two sites. No significant change with side (medial or lateral) was found. A small decrease in the chondroitin sulphate to keratan sulphate ratio without change in the total amount of hexosamines with age was found on the caudal condylar summits in these adult individuals below the age of 50.


The linkage of corneal keratan sulfate to protein.

The linkage of corneal keratan sulfate to protein has been investigated. After exhaustive digestion of bovine corneas with papain and pronase, a product was obtained in which aspartic acid was the predominant amino acid and constituted 59% of the total amino acids. A carbohydrate-protein linkage fragment was isolated from this preparation by a relatively simple procedure involving the following steps: (1) partial acid hydrolysis, adsorption of glycopeptides and other cationic material on Dowex 50-X2 (H+) and elution with 0.25 M HCl: (2) paper electrophoresis of the eluted fraction at pH 6.5 and pH 1.9; (3) paper chromatography; and (4) final purification by column chromatography on Aminex A"-5 resin. The structure of the linkage fragment was established as 2-acetamido-1-(L-beta-aspartamido)-1,2-dideoxy-beta-D-glucose (Asn-GlcNAc). Evidence for this structure was obtained from qualitative and quantitative analyses as well as from the migration characteristics in several chromatographic anc electrophoretic systems. Further support for the identity of the isolated compound was provided by treatment with beta-aspartyl N-acetylglucosyl-amine amidohydrolase which specifically cleaves Asn-GlcNAc or asparaginyl-oligosaccharides. It is concluded that corneal keratan sulfate is bound to protein via a N-glycosylamine linkage between N-acetylglucosamine and asparagine: this type of linkage is common to many glycoproteins.


The recovery and characterization of acid glycosaminoglycans in normal human urine. Influence of a circadian rhythm.

A method is described for the recovery of acid GAG (AGAG) as solid Ba salts from relatively small volumes (100+/-50 ml) of normal adult human urine. It is pointed out that (a) the amount recovered, (b) the molecular characteristics (i.e. molecular weight and charge density), (c) the ionic strength at which the AGAG are recovered and (d) the "critical electrolyte concentrations" of the AGAG-precipitating reagent (or anion exchanger) are inseparably linked, and that different fractions are obtained by variations in any parameter. Our product is defined as that fraction of heterodisperse material precipitated at room temperature by cetyl trimethylammonium bromide from urine diluted with 3 volumes of 0.05 M sodium acetate buffer pH 5.7, in the presence of asbestos powder. It is less degraded than other AGAG remaining in the urine supernatant. The product contained three electrophoretically separa0le components, characterized by susceptibility to pronase digestion, infrared and ultraviolet spectroscopy, hexosamine identification, tests for uronic and sialic acids, and the "critical electrolyte concentrations" of their Alcian blue complexes, as "chondroitin sulphate", "heparan sulphate" and acid glycoprotein. The method is simple, rapid and suita0le for use on multiple samples. The product is directly usable in electrophoretic analyses, etc. Using this method AGAG was recovered from the urine of nine males and five females, aged 25-43, collected during a 24-hr cycle, in six 4-hr aliquots. The ratios of "chondroitin sulphate" to "heparan sulphate" quantitated from the electrophoretic strip, showed a marked peak in every subject at 18-22 hrs.


Characterisation of the major CNBr-Derived peptides of porcine type II collagen.

As part of a general study on the matrix macromolecules of pig cartilage, the CNBr-derived peptides of porcine type II collagen have been isolated from laryngeal cartilage and characterized. Type II was the only molecular type of collagen detected in laryngeal cartilage from 6-9 month old pigs. The major CNBr-peptides of this collagen were prepared by ion exchange and molecular sieve chromatography and characterized by SDS-polyacrylamide disc electrophoresis and amino acid analysis. Six peptides were recovered in high yield and were shown to be closely homologous to similar peptides previously recovered from bovine and human type II collagens. The largest peptide alpha1(II)CB10 appeared by SDS-disc electrophoresis to be slightly larger than previously reported. The amino acid composition of alpha1(II)CB10 supported this finding of a higher molecular weight.


The behavior of (14C)-D-penicillamine in collagen metabolism.

The most important effect of penicillamine on collagen metabolism is the reduction of collagen crosslinking. However, even after long time application of penicillamine, collagen is crosslinked to a certain degree. After intravenous injection of a trace dose of (14C) labelled D-penicillamine it can be determined that this substance is rapidly bound to neutral salt soluble, acetic acid soluble and urea soluble collagen fractions and to a lesser extent to insoluble collagen as well. The amount of penicillamine which binds to any of the collagen fractions depends on the turnover rate. When different tissues are compared, penicillamine seems to have the greatest affinity to tissues with a high collagen turnover. Further studies of neutral salt soluble collagen by CM-cellulose chromatography revealed a stable linkage of penicillamine to collagen alpha chains.


Distribution studies of proteoglycan aggregates by ultracentrifugation.

The tendency forcartilage proteoglycans to aggregate or disperse in ultracentrifugation at high ionic strength is influenced by cations. Guanidinum, Li+ and Ca++ promote disaggregation and Na+, K+ and Cs+ support aggregation. The process of disaggregation is fundamentally the separation of low density from high density proteoglycans, the density being an inverse function of the protein content. PPL 5 pellet formation is a packing or aggregation phenomenon occurring well below the isopycnic density of proteoglycan which accounts for its difference from the isopycnic density equilibrated complex. Cations which promote disaggregation result in shifts in distribution of the proteoglycans on equilibrated density gradients. Comparisons of the desulfated products of the high density aggregated and disaggregated complexes and of more selective high and low density proteoglycan fractions clearly show the heterogeneity of proteoglycans related to protein content. It is suggested that the nature of the variable population of proteoglycan macromolecules permits its existence as a self aggregating system.


The effect of calcium on the macromolecular properties of heparan sulfate.

Heparan sulfate from human aorta has been subjected to a physico-chemical analysis in buffers of physiological ionic strength containing either sodium chloride or calcium chloride. A molecular weight of 50,000 was obtained both in sodium and calcium solutions by sedimentation equilibrium and from sedimentation and diffusion coefficients. The values obtained for So20,w in sodium and calcium chloride solutions were 2.28 X 10(-13) and 2.70 X 10(-13) sec, respectively, and the corresponding values for Dl20,w were 2.7 X 10(-7) and 3.1 X 10(-7) cm2/sec, respectively. Diffusion coefficients calculated from data obtained by gel chromatography were in excellent agreement with those determined by conventional techniques. The results indicated that the molecule contracts in the presence of calcium, presumably due to an increased binding of counter-ions with a concomitant decrease in charge density. Circular dichroism spectra above 200 nm, where the substituted amino group contributes to the absorption, gave no indication of a conformational change in the polysaccharide upon transformation from the sodium to the calcium salt. When the polysaccharide was dissolved in a salt solution, physiological both in ionic strength and in sodium to calcium ratio, it sedimented as the sodium salt. The sedimentation coefficient, the diffusion coefficient and the apparent molecular weight all displayed a concentration dependence. This dependence was much less in calcium chloride than in sodium chloride in determinations of molecular weight. Therefore there are advantages in performing molecular weight determinations of glycosaminoglycans in calcium solutions.


A comparative trial of azapropazone and ketoprofen in the treatment of acute backache.

A double-blind trial of azapropazone (300 mg. 4-times daily) and ketoprofen (50 mg. 4-times daily) was carried out in 50 patients with acute backache sufficiently severe to necessitate hospital admission. Of 39 patients who completed the full 3-weeks' study period, 18 preferred azapropazone therapy, 10 preferred ketoprofen, and 11 showed either preference for the intermediate placebo period or no preference at all. Ten patients suffered from sufficiently severe side-effects with ketoprofen to necessitate their withdrawal from the trial. There were no similar episodes of withdrawal occurring during the azapropazone period.


Inhibition of mitosis and tritiated DNA precursor uptake in human leucocyte cultures by rat lymph node extract.

A water soluble extract was prepared from the lymph nodes of rats of different age groups. This extract, when added to human peripheral blood leucocyte cultures, inhibits DNA synthesis and mitosis when added in different concentrations for varying lengths of time in culture. Also, it was observed that the extract from old rats was not as effective as that from younger animals.


Familial transmission of a (21q22q) translocation.

A large family is described in which a (21q22q) Robertsonian translocation is segregating through three generations. The assessment of the risk of a translocation carrier producing an offspring with Down's syndrome is calculated from the data in this family and eight others reported in the literature. The risk when the translocation carrier is a female is approximately 6 in 100, or 0.06. For the male translocation carrier the risk can only be guessed, since the patients with Down's syndrome born to these parents were probands. The risk for Down's syndrome from the combined data of male and female translocation carriers in 3 is 100, or 0.03.


Method for the quantitation of steroids in umbilical cord plasma.

A method for simultaneous quantitation of nine steroids in cord plasma is described which consists of Amberlite XAD-2-column chromatography at constant temperature of 45 degrees C, enzyme hydrolysis with beta-glucoronidase/aryl sulfatase, addition of five radioactive internal standards, ethyl acetate extraction, thin layer chromatography and quantitation by gas-liquid chromatography after trimethylsilyl ether derivative formation. Reliability criteria are established and the following steroid concentrations found: progesterone, 132.1 +/- 102.5 mug/100 ml; pregnenolone, 57.3 +/- 45.7 mug/100 ml; dehydroepiandrosterone, 46.5 +/- 29.4 mug/100 ml; pregnanediol, 67.5 +/- 46.6 mug/100 ml; 16-ketoandrostenediol 19.8 +/- 13.7 mug/100 ml; 16 alpha-hydroxydehydroepiandrosterone, 126.3 +/- 86.9 mug/100 ml; 16 alpha-hydroxypregnenolone, 78.2 +/- 56.5 mug/100 ml; androstenetriol, 22.2 +/- 17.5 mug/100 ml and oestriol, 127.7 +/- 116.9 mug/100.


Metabolism of 14C-dehydroepiandrosterone in female adipose tissue and venous blood.

The metabolism of dehydroepiandrosterone in female adipose tissue and venous blood in 11 patients was studied by a double isotope method which is described in detail. The main metabolite isolated, identified and quantitated was androstenediol. The conversion rate of dehydroepiandrosterone to androstenediol for adipose tissue ranged between 3.32-14.28% (X = 7.47 +/- 3.34 SD) and for venous blood between 2.88-9.60% (X = 5.84 +/- 1.80 SD). The values found for androstenedione and testostrone did not differ from the control experiments. Oestrone and oestradiol could not be detected. The contribution of the peripheral steroid metabolism to the pool of androgenic steroids is discussed.


Effect of methysergide, a blocker of serotonin receptors, on plasma prolactin levels in lactating and ovariectomized rats.

The effect of methysergide (MES, 2.5 mg/100 g body wt), a serotonin antagonist, on prolactin release has been studied in lactating and ovariectomized rats. MES caused significant increases in prolactin release in both animals. Studies in ovariectomized, hypophysectomized rats indicate that this effect is not due to a decrease in the peripheral metabolism of prolactin. In vitro incubations of anterior pituitary fragments with MES failed to demonstrate any increase in prolactin release, suggesting that MES does not act directly on the anterior pituitary. Parachlorophenylalanine (PCPA; 32 mg/100 g body wt) decreased brain serotonin levels in ovariectomized rats 5, 24, and 70 h after its administration, yet did not alter plasma prolactin levels. L-tryptophan (6.3 mg/100 g body wt) given 1 and 1 1/2 h prior to sacrifice increased brain serotonin levels, yet did not affect plasma prolactin levels. Neither PCPA nor L-tryptophan altered MES-induced prolactin release. In lactating rats, suckling caused marked increases in plasma prolactin levels, an effect completely abolished by the administration of MES to the mother rats 3 1/4 h prior to suckling. However, MES-induced prolactin release was not altered by prior treatment with MES, either in lactating or ovariectomized rats. Others have shown that suckling releases prolactin through an excitatory serotonergic mechanism. Therefore, the failure of suckling to release prolactin in MES-pretreated rats suggests that MES can block brain serotonin receptors. However, the ability of methysergide to release prolactin in rats with serotonin receptors presumably blocked, suggests that the serotonin receptor-blocking and the prolactin-releasing actions of MES are not related.


Influence of adrenalectomy, castration, and light cycle on steroid hydroxylase activity in the adult male rat liver.

The contribution of the adrenals, the testes, and a light/dark cycle to the regulation of hepatic steroid hydroxylases was studied in vitro by measuring transformation rates (nmol/mg protein/min) of dehydroepiandrosterone (DHA). Normal and adrenalectomized male rats were kept in 12 h light:12 h dark (12L:12D) or in constant light (CL) for 1 week. Normal and castrated rats were kept in 12L:12D for 3 weeks. Rats were killed at 4-h intervals over a 36-h period, hepatic microsomal fractions were incubated with [4-14C]DHA, and C-7 and C-16-oxygenated products were quantitated. The overall mean rate of 7alpha-hydroxylation decreased in normal rats exposed to CL. Rates of both 7alpha- and 16alpha-hydroxylation were significantly greater in normal rats than in the respective group of treated rats, except for the 7alpha-hydroxylase in castrated rats. These results indicate that 16alpha-hydroxylase activity is both gonadal- and adrenal-dependent, whereas 7alpha-hydroxylase activity is adrenal-dependent only. Both enzymes are sensitive to light/dark modulations, although a circadian rhythm could not be conclusively established.


Poly(A) synthesis in T2L phage-infected Escherichia coli. A combination of polynucleotide phosphorylase and ATPase.

In crude extracts of T2L phage-infected Escherichia coli cells an enzyme activity was found that produced poly(A) from ATP as substrate. Purification of the extract led to the isolation of two enzymes, a polynucleotide phosphorylase and an ATPase. The polynucleotide phosphorylase possessed the same properties as the well-known enzyme from uninfected cells and its molecular weight was about 265 000. The ATPase was purified to over 90% purity; its molecular weight was estimated to be about 165 000 with three subunits of 55 000. The characterization of this enzyme showed that it was different from any ATPase known so far. Mg2+ cannot be replaced by Ca2+, as it can from the membrane-bound ATPases. The only product yielded by the enzyme was ADP; it was very specific for ATP, other ribonucleotide triphosphates being practically unaffected. The rate of ATP splitting was found to be very high, the turnover number being 2.51 X 10(4) min-1 at 37 degrees C. Even at 0 degree C the enzyme was still active. The optimal assay conditions for ATPase turned out to be very similar to those of polynucleotide phosphorylase. Thus the combination of the two enzymes very efficiently produced poly(A) from ATP. In this combination the polynucleotide phosphorylase was the rate-limiting enzyme, since its turnover number was about 40 times lower than that of the ATPase. The evaluation of a variety of properties of the poly(A)-synthesizing constituent found in the crude extracts led us to conclude that this activity arises from the combined action of ATPase and polynucleotide phosphorylase, and is not due to a poly(A) polymerase.


Amino-acid sequence of the cyanogen-bromide fragment from human plasminogen that forms the linkage between the plasmin chains.

The complete amino acid sequence of a cyanogen bromide fragment (122 residues) obtained from plasminogen is described. This fragment forms the overlap between heavy (A) and light (B) chains of human plasmin. The particular arginyl-valyl bond cleaved in the second step of the activation process is shown to be Arg98-Val99 in this fragment. This site is not very similar to the one in the NH2-terminal part of the molecule (Arg68-Met69). Remarkable homologies with the 'triple loops' ('kringle structures') found in the non-thrombin part of prothrombin are demonstrated. Homologies occurred during evolution of this chain.


Tightness and orientation of vesicles from guinea-pig kidney estimated from reactions of adenosine triphosphatase dependent on sodium and potassium ions.

In order to study the "sidedness" of the ligands of the Na+, K+-ATPase in the phosphorylation from [32P]ATP, tight vesicles were prepared from guinea pig kidney and partially purified by a two-stage sucrose and Ficoll gradient centrifugation procedure. These vesicles were derived presumably from plasma membrane fragments resealed after the initial disruption of the cells during homogenization. Tightness of the vesicles was estimated according to activation by the nonionic detergent, Triton X-100. Treatment with Triton X-100 increased both the activity of the Na+, K+-ATPase and its Na+-dependent phosphorylation from [32P]ATP at least three-fold. Activation of both functions also appeared when the vesicles were shocked osmotically. These results suggest that the preparation contains a major population of tight normal vesicles (approximately 75%) in which the phosphorylation site faces the intravesicular solution. In the response to ouabain breakdown of the phosphoenzyme was inhibited in vesicles treated with Triton X-100 but not in intact ones as if ouabain could not get to its binding site. Correspondingly in phosphorylation from ATP pretreatment with ouabain in the presence of inorganic phosphate produced less inhibition in intact vesicles than in those disrupted with Triton X-100 beforehand. These data suggest the presence of an everted vesicle fraction in the preparation (approximately 20%). Apparently only a small fraction of the vesicles was leaky. In the everted vesicles the action of K+ on the phosphoenzyme was slow. In order to accelerate the dephosphorylation in intact vesicles as effectively as in disrupted ones, K+ had to be added before the start of phosphorylation. This supports the view that K+ was acting from the side of the membrane opposite to that where the gamma-phosphoryl group was accepted from ATP.


A theory concerning the course of events in angina and myocardial infarction.

Evidence is discussed which suggests that coronary arterial thrombosis need not be the primary event in myocardial infarction. If this is so, it is necessary to consider the initial changes which might occur in the myocardium rather than the effects of experiments in which the coronary arterial supply has been suddenly cut off. Results of recent investigations on patients suggest that, during the anginal attack, there is an inability of the affected myocardium to relax and a diminution in the flow of blood to the region. The former implies an increased myofibrillar concentration of calcium ions, the latter may be explained by the mechanical compression of small intramyocardial blood vessels. It is argued that these two factors may initiate vicious circles which could lead to the death of myocardial cells and thus to myocardial infarction.


Influence of 1,3-butanediol on blood glucose concentration and pancreatic insulin content of streptozotocin-diabetic rats.

Male rats were made diabetic by intravenous administration of 75 mg/kg of streptozotocin and were fed, via a pair-feeding regimen, high-fat diets +/- 1,3-butanediol (BD) at 13.5 and 27% of the dietary calories for 30 days and 31 days, respectively. 1,3-Butanediol was added to the diets primarily as a replacement for fat. Food consumption and rat weight were recorded daily. Whole blood glucose concentrations were determined weekly. At sacrifice, liver, pancreas and epididymal fat pads were excised and blood samples were collected. Liver was analyzed for protein and lipid; pancreas was weighed and analyzed for insulin; fat pads were weighed and discarded; and blood was analyzed for glucose and lipid. The 13.5% BD diet increased the beta-hydroxybutyrate, acetoacetate and cholesterol concentrations, decreased the glucose concentration in blood, and increased the insulin content of the pancreas. The BD diets did not affect the concentrations of phospholipid, triglyceride, cholesterol and fatty acid in the liver; fatty acid concentrations in the blood; or the epididymal fat pad weight. The results suggest that BD produced a slight amelioration of the diabetic condition, which may have resulted from an increased capacity of the pancreas to synthesize insulin. In addition, the data provide further evidence suggesting that in the rat BD is oxidized to the ketone bodies, beta-hydroxybutyrate and acetoacetate.


Ovarian delta-5-3-beta-hydroxysteroid dehydrogenase in aging rats.

Ovaries of rats 1, 2, 4, and 18 months old revealed significant delta-5-3-beta-hydroxysteroid dehydrogenase (3beta-OHSD) activity when dehydroepiandrosterone (DHA), pregnenolone, and 16-dehydropregnenolone served as substrates. No enzyme activity was evident in ovaries 4 and 18 months of age when the substrates 17alpha-hydroxypregnenolone, pregnenolone sulfate, and dehydroepiandrosterone sulfate were used. A marked increase in enzyme activity occurred in the granulosae of mature vesicular follicles at 18 months of age. DHA, pregnenolone, and 16-dehydropregnenolone provided similar activities. On the other hand, interstitial enzyme activity exhibited an aging decline with DHA and 16-dehydropregnenolone but increased with pregnenolone. Corpora lutea were less evident in aging ovaries but did exhibit strong 3beta-OHSD activity when DHA was used. Pregnenolone use by corpora lutea was sharply increased in aging rats but no change occurred with 16-dehydropregnenolone. Thus aging was associated with changes in 3beta-OHSD distribution and use of steroid substrate; a change in function is suggested.


Spring clip tubal occlusion: a report of the first 400 cases.

Between September 1972 and March 1974, laparoscopic tubal sterilization was performed on 394 patients with the use of a spring-loaded plastic clip under local analgesia and on an outpatient basis. Intravenous fentanyl and infra-umbilical infiltration and tubal spray with 1% lidocaine were found to be very acceptable and effective forms of analgesia. The average time spent by a patient in the hospital was 4 hours, 26 minutes. There was no major immediate complications apart from vasovagal reactions in 34 patients. Up to June 30, 1974, six patients in the series had become pregnant, not as a failure of the clip per se, but as a result of improper application of the clips. This seems to be a safe, simple, and effective method of female sterilization with great potentials of reversibility.


Sterilization by spring clip: a report of 1000 cases with a 6-month follow-up.

In September 1972, clinical trials of a spring-loaded clip for laparoscopic sterilization were begun and extended to a number of centers in the United States and overseas. As of March 1974, more than 1000 patients had undergone the procedure, usually performed under local anesthesia in an outpatient setting, with no fixed contraindications. Complications and pregnancy rates based on a preliminary 6-month follow-up are presented. Complications due to application of the clip appeared to be limited to postoperative cramps lasting 24 to 48 hours (26% of the patients). No ectopic pregnancies were reported. Pregnancies, when corrected for unsuspected pregnancies and misapplication of the clip, occurred in 2 of these first 1000 patients. The difficulties and relative contraindications learned from this unselected series and the advantages over electrocagulation  techniques are discussed.


Peripheral, ovarian, and adrenal vein steroids in hirsute women: acute effects of human chorionic gonadotropin and adrenocorticotrophic hormone.

In an attempt to localize the source of excessive andogens in hirsute women, various steroids were determined in peripheral blood, left ovarian venous blood, and left adrenal venous blood, using radioimmunoassay techniques developed in our laboratory. Peripheral serum levels of dehydroepiandrosterone, androstenedione, 5alpha-dihydrotestosterone, testosterone, progesterone, and 17 alpha-hydroxyprogesterone were elevated in several hirsute patients, compared with those of a group of normal, cycling women of the same age. There was direct evidence to show that the ovaries and/or adrenals were secreting androgens in these hirsute patients. Saline infusion brought about minor changes in peripheral and left ovarian vein steroid levels; however, episodic fluctuations occurred in the adrenal vein samples. Adrenocorticotrophic hormone infusion brought about dramatic increases in adrenal vein steroid concentrations, while ovarian vein concentrations were unaffected, compared with peripheral steroid levels. Human chorionic gonadotropin (HCG) infusion brought about increases in left ovarian vein steroids in some patients; however, those patients with corpus luteum on the right ovary or under chronic ovarian suppression did not respond to HCG. HCG was ineffective in altering adrenal steroid secretion. It was concluded that selective venous cannulation may be of some use when the ovary is the site of excessive androgen secretion. However, care must be taken in interpreting results when a corpus luteum is present in the contralateral ovary. Interpretation is difficult when the adrenal is suspect, because of the pulsatile nature of adrenal secretion or the possibility of stress-induced secretion.


Elementary particles in isolated rat liver and spleen nuclear membranes.

Elementary spherical particles similar to those described in the mitochondria are found in isolated rat liver and spleen nuclear membranes. The particles are characterized by electron microscopy of sections and preparations negatively stained with phosphotungstic acid or with the ATPase histochemical reaction product. The particles exhibit adenosine triphosphatase activity and have a mushroom-like shape with a sphere about 85-90 angstrom in diameter attached to inner face of the inner nuclear membrane by a stalk about 50 angstrom in length and 35 angstrom in diameter. It is supposed that these particles, just as analogous particles localized at the inner mitochondrial membrane, are involved in the coupling of oxidation and phosphorylation in the nuclear envelope.


Psychological disorders in crippled children. A comparative study of children with and without brain damage.

A detailed standardised study was made of all crippled children aged between five and 15 years and of normal intelligence on the local-authority lists of handicapped children in three London boroughs. Psychiatric disorder was twice as common in children whose crippling was due to cerebral disease or damage rather than some peripheral lesion. As the groups were well matched in terms of physical incapacity and social background, it was concluded that brain damage was responsible for the children's increased vulnerability to emotional problems. Brain damage was also associated with a marked increase in reading difficulties and a lowering of intelligence within the normal range. Psychiatric disorder was found to be related not only to cerebral injury but also to various types of family disturbance. It is concluded that emotional and behavioural disturbance stemmed from both an increased biological vulnerability and psychosocial hazards.


Adoption and foster care of handicapped children in the United States.

This paper reviews the current policies and programs of adoption and foster care of handicapped and other "hard-to-place" children in the United States. The results of a survey of 42 States revealed a variety of approaches and attitudes towards recruitment of parents and placement of handicapped children. It is concluded that successful programmes related directly to careful selection of families, to educating the parents and to adequate staff support.


The effects of thymidine and its analogues on the differentiation of antibody-forming cells.

Thymidine (Tdr) and its analogues 5-fluorodeoxyuridine (FUdr) and 5-bromodeoxyuridine (BUdr) inhibited the development of antibody-producing cells (PFC) in cultures of explants of rabbit spleen stimulated with sheep red cells in vitro. They were effective in this respect, even at concentrations too low to inhibit cell division. The results indicated that Tdr had produced its effects by a selective action on DNA and RNA synthesis. In view of what is known about the mechanisms of action of Tdr and its analogues to interfere with DNA-dependent events in cells, these results supported the idea that DNA synthesis, unrelated to the needs for cell division, was required for the development of antibody-producing capacity by PFC in these cultures.


[Isolated tricuspid insufficiency of traumatic type. Report of a case and revision of the literature (author's transl)].

A case of isolated tricuspid insufficiency is reported. The lesion, that at surgery resulted to be an anterior valve leaflet detachment, was well tolerated for a long time. After 23 years from the trauma the patient was successfully treated by surgical correction. The clinical features, compared to the 33 cases reported in the literature, demonstrated that the clinical course was more dramatic in patients with papillary muscle rupture than in those where the lesion was located on the chordae tendineae and/or the valves. The physical findings are characteristic but not specific of the traumatic type of tricuspidal insufficiency. However, when these symptoms are found in a patient where a trauma has previously occurred, the correct diagnosis should consider this possibility. The ECG usually shows incomplete bundle branch block, and the chest roentgenogram reveals cardiomegaly. Furthermore, the blood pressure levels in the right chambers are nearly normal. Surgery is often indicated and seems to give very good results. Our patient markedly improved after surgical treatment. Electrocardiographic and vectorcardiographic evidence of right atrial and ventricular hypertrophy was reduced as well as the roentgenographic pattern.


The program of the cooperative study of the Veterans Administration.

A computer program for the automatic analysis of the electrocardiogram is described with details about wave recognition, measurements, and the calculation of the posterior probabilities of each diagnosis. The advantages and disadvantages of this multivariate statistical procedure are discussed. Classification statements result from separate analysis of the QRS-T complex, P wave, and ST segment. Criteria for inclusion of the patients forming the data base are given, with tables of measurements, means and standard deviation, as well as investigation of agreement between calculated and observed probabilities. Estimated error rates are given in the form of misclassification matrices, computed from large numbers of tracings collected by a Veteran Administration Cooperative Study, where the correct diagnoses are taken from clinical, laboratory, and autopsy information.


Unscheduled DNA synthesis in human lymphocytes treated with neocarzinostatin.

In an attempt to quantitate DNA repair induced by Neocarzinostatin, the incorporation of labeled thymidine into the human peripheral lymphocytes after exposure to the agent was studied by autoradiography. The exposure of lymphocytes to Neocarzinostatin for 60 min at the concentrations as low as 0.05 mug/ml caused a significant increase in the number of grains per cell within 2 hr of incubation with 10 muCi/ml of 3H-thymidine in the presence of 2mM of hydroxyurea. The number of grains increased with the increasing dose of the agent up to the concentration of around 2.5 mug/ml and then fell at higher concentrations. The extent of maximum incorporation induced by Neocarzinostatin was found to be almost comparable to that induced by 100 erg/mm2 of UV irradiation. Hydroxyurea or caffeine in the labeling medium showed little or no effect on the grain count, but acriflavine at 0.1 mM reduced the grain count by a factor of about 6. These results indicate that relatively high level of repair synthesis occurred in human lymphocytes after exposure to Neocarzinostatin and provide further evidence for the direct damage of cellular DNA induced by the antibiotic.


[Gynaecological laparoscopy (author's transl)].

The double puncture laparoscopy was abandoned in our department in favor of the single puncture technique. 187 diagnostic laparoscopies and 617 laparoscopic tubal ligations are reviewed. In 46% of the diagnostic laparoscopies, the findings were confirmed. In 40% of the diagnostic laparoscopies, normal findings were seen and in 11%, a different abnormal finding than suspected was found. In 3% of women, the findings could not be elucidated by laparoscopy. None of the 617 patients who underwent laparoscopic tubal sterilization have become pregnant. In 5% of the patients (0.6%) complications occurred. In 3 cases a laparotomy became necessary. In 2 cases there was bleeding from the fallopian tube or the greater omentum and in 1 case there was a trauma to the intestine. Laparoscopic tubal sterilization by the single puncture technique and coagulation without division of the tube only, is considered to be the lowest risk method of the laparoscopic tubal sterilizations.


[Metastasising carcinoma of the renal pelvis after Thorotrast pyelography].

A carcinoma of the renal pelvis was diagnosed in a female patient 42 years after a retrograde Thorotrast pyelogram. The patient died as a result of the carcinoma at the age of 60. The literature is reviewed. As a result of this case, the need is stressed for early recognition of Thorotrast depositis in the kidney. In this case it was demonstrated by autoradiography. Appropriate treatment is discussed.


[Catheter embolisation of a bleeding renal hamartoma, with a contribution to its angiographic diagnosis].

Hamartomas of the kidney, often associated with tuberous sclerosis, have a typical angiographic appearance. Despite the similarity of the angiographic signs with malignant renal tumours, it is possible to differentiate these, as was shown in two patients which we have documented. If there is massive bleeding, fibrin embolisation carried out at the end of a diagnostic angiogram may lead to occlusion of the vessels and stop the bleeding. This was demonstrated in one of our patients.


[Spontaneous peri-renal haematoma as a complication of chronic pyelonephritis].

Three cases with spontaneous peri-renal haematomas, a rare complication of chronic pyelonephritis, are described. The clinical features and diagnosis of this condition are discussed with reference to these cases and the literature is reviewed.


[Calcium "milk" in the kidney. (Five cases with hydronephrotic kidney)].

A study of the literature has revealed 11 cases of calcium "milk" in hydronephroses. We have been able to observe five further cases in a period of 14 months. The criteria annunciated by Murray are stressed, since they will raise a suspicion of the presence of calcium "milk" on views taken with a vertical x-ray beam. In three patients, the cause of the hydronephrosis was a stone in the pelvi-ureteric junction. In the other two cases, there was a stenosis below the pelvis. Murray's criteria permitting identification of calcium "milk" with a vertical beam were analysed. Not all his criteria were found to be reliable.


[Renal scintigraphy with 99mTc-penicillamine].

By application of 99mTc-Penicillamine in renal scintigraphy excellent scintigraphies of the kidneys, especially by using the gamma-camera, can be obtained, even in case of damaged renal function, up to an increase of creatinine in serum of about 7 mg %.


[An approximation for improving the accuracy of compartmental analysis after intravenous 133Xe].

Perfusion of transplanted kidneys is studied in our clinic by intravenous injection of 133Xe with subsequent graphic compartmental analysis of time-activity curves. Where there is poor perfusion of the renal cortex, it is not possible to obtain a half-value time for the first compartment--which is the significant measure for specific blood flow -- since few points are available for the tail end of the curve. A correction is described which produces an approximation to a bolus, based on the time-activity curve of the expired air. If this is applied to the proximal part of the time-activity curve, with respect to background activity, compartmental analysis can be performed with a higher degree of accuracy in the calculation of renal perfusion.


[Scintigraphic findings in splenic vein thrombosis].

After a brief review of the literature concerning scintigraphy of the spleen, its vascular system is described. The presence of numerous anastomoses makes it likely that occlusion of the splenic vein will usually not lead it any parenchymal damage. In five patients with angiographically confirmed spenic vein thrombosis it was shown that radio-colloid scans always demonstrate an enlarged spleen. Poor colloid uptake was thought to be due to a venous reflex mechanism leading to a reduction in the arterial blood supply. The statement found in the literature, that the spleen cannot be shown scintigraphically if the vein is thrombosed, has not been confirmed by us. We tend to the view that the apparent absence of a splenic scan was due to geometric causes.


[Changes in vascular topography after surgery in the upper abdomen].

In 20 patients who had had upper abdominal surgery (18 gastric resections, one removal of spleen and one chole-dochoduodenostomy), it was necessary to perform abdominal angiography. Changes in the coeliac trunk and superior mesenteric artery due to the surgery were analysed. Displacement of vessels and changes in calibre were the most common findings. These changes must be kept in mind, particularly in the diagnosis of tumours.


[Changes in serum osmolarity after high contrast doses during angiography].

Analysis of changes in osmolarity of the serum after various doses of hypertonic radiographic contrast media, following a single injection, showed an initial rise of about 35 m/mosm/1, followed by a rapid fall in osmolarity with a subsequent short hypo-osmolar phase. With repeated injections, similar changes are found if the intervals between the injections are long enough. Following repeated injections over a short interval, there may be a more prolonged phase of hyperosmolarity. In this phase of delayed dehydration, there may be clinical complications. Possible forms of treatment are discussed.


[Slow infusion cholangiography in jaundice. An experimental study in the dog].

56 intravenous cholangiograms were performed in eleven dogs with either advanced hepatic damage or complete common bile duct obstruction. If radiographic visualization of the biliary system did not occur within 8 hours after a 30 minute infusion with 0.6 ml/kg iodipamide 52%, the result could not be improved by just extending the infusion time to 2 and 6 hours. However, a slight improvement was obtained, when the iodipamide dose was increased up to 1.8 ml/kg. Nevertheless we do not recommend a massive increase of the dose in the icteric patient, because it is likely that the risk of toxic side effects increases with an increasing iodipamide dose.


[Slow infusion cholangiography in patients with jaundice].

Thirty patients with jaundice and serum bilirubins above 4 mg.% were examined by slow infusion cholangiography (40 ml. (20 g.) Ioglycamid in 500 ml. 0.9% NaC1 in ten hours). Diagnostic results were achieved in 38%; in 15 patients with extra-hepatic obstruction, the success rate was 13%, whereas in disease of the liver parenchyma, the success rate was 60%.


[Radiological aspects of esophageal intramural diverticulosis].

The 9th observation of esophageal intramural diverticulosis is reported. The findings are compared with those of former publications. Clinically dysphagia is a leading symptom. Suction biopsy proved diverticulosis being the correct interpretation for radiological findings. The etiology is discussed. The change between symptom-free intervals with diverticulosis only and episods of dysphagia by secondary inflammation (reflux esophagitis, moniliasis, stenosis) stresses the importance of such secondary complications. These inflammations can hide the real diagnosis for a long time. The radiological findings are the key for diagnosis, however, they can be very small for years. During this period a diagnosis can only be achieved by knowledge of the disease and the skilful search for early symptoms.


Congenital diaphragmatic hernia (Bochdaleck) with special reference to the prognostic influence of congenital cardiac and gastro-intestinal anomalies.

Of 29 patients with hernia Bochdaleck there were 11 in the non surgical group and 18 in the surgical group. 14 were operated in the first 24 hours of life, and 4 patients after that time. No patients in the non-surgical group survived. In patients who were operated upon in the first 24 hours of life, 5 survived, and after that time 3. In all cases the transthoracic approach was used. The investigation did not permit any safe judgement of the preoperative treatment. On these conditions multiple anomalies combined with prematurity and intrauterine asphyxia had some influence on, whether a patient lived so long, that an operation could take place. Gastro-intestinal anomalies had a certain prognostic significance in the postoperative period. In cured patients the severity of a complicating cardiac failure could be of some importance.


[Radiological and clinical aspects of resection of the cardia].

Following resection of the cardia, problems may arise during two phases; in these, clinical and radiological examination may be required. Immediately after the operation one is concerned particularly with checking the incision or the extent and localisation of suture abnormalities. In the late post-operative period the problems are different. In addition to a description of the anatomy at the proximal and distal ends of the stomach, it is necessary to estimate functional conditions, such as passage through the anastomosis and the pylorus, the reservoir function of the gastric remnant and gastro-oesophageal reflux. An important task is to demonstrate tumour recurrence. For this purpose endoscopic, biopsy and cytological examinations are required.


[Neuromas and myomas of the stomach].

Seven non-epithelial benign tumours of the stomach are described. The clinical and radiological features are compared with those described in the literature. Since it is not always possible to be certain of the benign nature of the lesion, and since they may give rise to life-threatening bleeding, resection is indicated in every case. High position of the tumour tends to indicate a benign lesion.


[Roentgen kymographic investigations of gastric peristalsis after intravenous injection of caerulein].

Caerulein is a decapeptide which combines the effects of gastrin and cholecystokinin-pancreozymin. It was injected intravenously in doses of 10 to 40 nanoponds caerulein per kilopond bodyweight in 37 patients and roentgen kymography of the stomach carried out before and after the injection. In 89% of the 36 cases which could be evaluated, it increased antral peristalsis. Amplitude and frequency were increased and the periodicity decreased correspondingly. The effect was more marked than that of pentogastrin, presumably because caerulein dose not affect duodenal acidity. The changes in motility therefore correspond with those produced by serotonin and cholecystokinin-pancreoxymin. Control examinations carried out under identical conditions, but without caerulein, showed no change in antral peristalsis.


[The value and limitations of ultra sound in the follow-up of patients with urological tumors].

The application of ultrasonic diagnosis in the care of patients suffering from tumors of the urogenital tract is discussed. The indication for demonstration of enlarged paraortic and lumbar lymphnodes is emphasized. Of special value is this new diagnostic tool for X-ray treatment planning and for the evaluation of the therapeutic success.


[The value of sonography in the diagnosis of renal space-occupying lesions].

1. Sonography achieves an accuracy of 93% (n = 55/59) in the localisation of space-occupying lesions in the kidney. Since the method is somewhat laborious, ultra sound, in the form of a slow B-scan, cannot be regarded as an alternative to radiology with an accuracy of 97% (n = 57/59). 2. For a pathological diagnosis, particularly in the distinction between tumours and cysts, sonography, with an accuracy of 98% (n = 54/55) is markedly better than radiology with an accuracy of 50% (n = 21/42). Sonography is approximately equal to angiography, which has an accuracy of 97% (n = 69/71). 3. Sonography is useful and is indicated for: a) the investigation of non-functioning kidneys, b) patients in whom radiographic contrast media are contra-indicated, and c) for puncture of renal cysts. (F. St.).


[The value and use of percutaneous trans-thoracic needle biopsy in the diagnosis of intra-thoracic disease. Technique, indications, results and complications].

Between 1961 and 1974, 5,300 percutaneous, trans-thoracic needle biopsies under image intensifier control were carried out in the Karolinska Sjukhuset, Stockholm. These involved, 2,726 patients with localised intrathoracic disease. The indications, results and complications are discussed. Approximately 15% of cases were suspected of malignancy and showed peripheral infiltrates varying from 0.04 to 2 cm. diameter. Needle biopsy proved to be technically simple and relatively safe; it permits early diagnosis of bronchial carcinomas and solitary metastases with a high degree of accuracy and with few complications. It is assumed that, as a result, the prognosis in a large group of patients with bronchial carcinomas in the pre-clinical stage may be markedly improved.


[Endobronchial brush biopsy].

Bronchial brushing under fluoroscopy control is used in the diagnosis of pulmonary lesions which are beyond reach of the bronchoscope. A small nylon or steel brush is passed through a bronchial catheter and tissue material from the lesion is taken for cytologic and bacteriologic studies. The technique described is easy to learn, relatively safe, is well tolerated by patients and gives good diagnostic results.


[Sequential scintigraphy of four different 99mTc human albumen preparations used for lung scanning].

The biological behaviour of four 99mTc-marked human albumen preparations, prepared by different methods, were studied by serial scintigraphy and excretion measurements. Three preparations consisted of particles of irregular albumen macroaggregates, the fourth was in the form of spherical albumen particles (microspheres). Forty-five patients with various pulmonary abnormalities were examined. The following parameters were used for comparison: half-life of pulmonary activity, changes in activity in liver and spleen as a measure for the formation of Tc-containing particle fragments, uptake in the thyroid and stomach and urinary excretion as an indication of the stability of the nucleid fixation. Despite marked differences in their biological behaviour, all four substances provide comparable information and lack of toxicity and are suitable for lung scintigraphy. Because of the rapid initial decline in activity in the lung, microspheres are more suitable for use with a gamma camera, whereas the other three preparations, in view of their long persistence in the lung can be conveniently used for the slower examinations with a conventional scanner.


Single ventricle: a new angiographic classification.

The authors recommend that the "single ventricle" be differentiated into two types. There are single ventricles of the left ventricular type, and others of the primitive type; in other words, the chamber is typically left ventricular or cannot be differentiated into left or right. In the latter case we are dealing with a primitive chamber. In addition there may be variations of the large vessels. Of 40 patients with a single ventricle of the left ventricular type, ten showed normal topography of the major vessels, 13 had a d-transposition and 17 an 1-transposition. In the d-transposition, the outflow track of the ventricle was anterior and to the right and the aorta arises in front of the pulmonary artery; in the 1-transposition and sub-aortic portion is anterior and to the left, the aorta is ventral and to the left of the pulmonary artery. In the discussion it is pointed out that the "single ventricle" must be included in the differential diagnosis of cyanotic, congenital heart disease. Angiographic confirmation of the "single ventricle" depends on the demonstration of an absent ventricular septum and of two separate atrio-ventricular valves.


[Complications of selective coronary angiography and the choice of contrast media].

The incidence of side effects of two contrast media containing different amounts of sodium (Urografin 76 and Conray 70) is compared in two groups of patients (totalling 133) under-going diagnostic examination by means of a cardiac catheter and selective angiocardiography. The following results were obtained: 1. Transient ischaemic reactions were observed in 4 patients of the Urografin group. Cerebral circulation was temporarily restricted in 2 patients, but there were no sequelae. One patient with a history of repeated cardiac infarction in whom the examination was the last hope with a view to coronary surgery died immediately after the angiocardiography. 2. In the Conray 70 group, ventricular fibrillation occurred 32 times in 22 patients after a total of 155 injections. Protracted asystole was observed in 2 patients. The explanation offered for the divergent findings from the use of the two contrast media is not the different anions--the iodine hosts--, but rather the difference in the amount of sodium contained in the preparations. It is concluded from the studies that contrast media which deviate to any great extent in their sodium content from the physiological range of the blood sodium concentration are unsuitable for angiocardiography.


[Laevo-cardiography for estimating the function of the left ventricle. II Mitral insufficiency].

1. The normal left ventricle is characterised by the relationship between end-diastolic volume and ejection fraction. (Reduction in the lateral projectional area by 50--60% during systole, as compared with the end-diastolic projectional area.)--2. The functionally sufficient left ventricle in the presence of haemodynamically significant mitral insufficiency shows and increasing end-diastolic measurement with increasing volume.--3. End-diastolic projectional area, up to 85 cm.2, and end-diastolic filling pressure, up to 20 mHg are associated, apart from odd exceptions, with normal contraction indeces, i.e. a normal ejection fraction. --4. Within these magnitudes of area and pressure, there is a demonstrable relationship between area/pressure quotient and contracture index. --5. Insufficiency of muscular contraction of the left ventricle in mitral insufficiency is characterised, with reduced normal or enlarged, end-diastolic areas and normal, or increased, end-diastolic filling pressure by an abnormal relationship between ejection fraction and end-diastolic volume (contraction index less than 50%). --6. In mitral insufficiency which is haemodynamically significant, some of the residual blood may be used as a supplement for ventricular filling.


[A modification of the Roher-Kahlstorf method for estimating rapid changes in cardiac volume].

A simple roentgenological procedure is described, by which it is possible to measure the heart volume and its alteration during early orthostatic adaptation.--Long distance films are taken in the p.a.-projection using a 70-mm-camera with a frequency of 2 to 4 exposures per second. The length and width and there by the plane of the heart are determined from these films. The mean horizontal depth diameter, that is necessary to calculate the volume of the heart can be gained by indirect means. There is no statistically significant difference between the heart volumes, that have been determined in 12 test persons by the method described, and the commonly used method. -- The alteration of the heart volume in 10 test persons were investigated during acute orthostatic stress. The heart volume, which was 693 +/- 24,8 ml before the orthostatic stress, decreased within 6,9 +/- 1,6 sec statistically significant by 16,5% An influence of the movements of the diaphragm on the heart volume determination could be excluded.


[Idiopathic, muscular, hypertrophic, sub-aortic stenosis].

The authors have seen 14 cases of idiopathic, hypertrophic, subaortic stenosis and they describe the pathology, the clinical and radiological diagnosis and the differential diagnosis of this condition. Their own findings are compared with those in the literature and substantial agreement is found. Some uncharacteristic clinical and radiological findings are associated with typical pressure curves in the left ventricle and marked abnormalities in the laevocardiogram. Variations in ventricular filling due to the difference in severity of the disease are discussed. The importance of a correct diagnosis is stressed, since the use of inotropic substance is this condition is contra-indicated.


[Deformities of the spine and ribs in embryologically related malformations of the heart with cyanosis].

Three cases of severe costovertebral deformities together with embryologically related cardiac malformations (Fallots Tetralogy and Truncus arteriosus communis) are presented. Two patients died because of their thoracic deformities and limited respiratory function. Relationships to similar bizarre veretebral anomalies are discussed. The cause is probably a teratogenic agent, acting between the fifth to eight embryologic week.


[The pharmaco-kinetics of angiographic contrast media with special reference to the extra-vascular spaces. Fundamental studies on dog for the characterisation of angiographic media. I The pharmaco-kinetics of various contrast media under conditions of constant infusion (balanced flow)].

The pharmaco-kinetics of angiographic contrast media in the extra-vascular space, which are largely unknown, were investigated experimentally in dogs. As part of a basic study, using radio-active contrast media, it was possible to determine the concentration and rate of elimination in practically all organs and tissues. Measurements were carried out first after prolonged infusion of the contrast under conditions of balanced flow, and secondly six hours after the end of the infusion. It was therefore possible to determine the inflow and loss of contrast medium in various organs, or organs systems. The most commonly used angiographic contrast media  in Germany were investigated. Their kinetic behaviour is largely identical, their pattern of distribution and elimination depended principally on the organ or tissue. A comprehensive discussion of the results of all the experiments will be given in the third article.


[How effective are tests for avoiding contrast reactions].

The frequency of known severe and fatal reactions after intravenous injections of biligrafin forte were compared during two periods. During the first period test ampules were provided; during the second period, the test ampules were no longer made. It was shown that the failure to perform tests did not result in any increase in the complication rate when compared with the period when test ampules were available. The problems arising from this comparison are discussed.


[Indirect (arterial) splenoportography and portography in intraphepatic block].

Morphology and flow dynamics in the lieno-mesentericoportal territory were investigated in 15 normal persons and 199 patients with pre/intra and intra-hepatic blocks. Indirect (arterial) splenoportography with selective catherization of the splenic artery provided angiograms varying between good and diagnostically useful in 98% of cases. With injection into the truncus coeliacus, this figure falls to 77%. The various types of collateral circulation are stated and discussed. The scope and limits of the methods are shown by comparative surveys between direct splenoportography and indirect splenoportography and portography, using 16 patients. As far as the chronological sequence of angiographical diagnostics in the intrahepatic block is concerned, the indirect method shall precede direct splenoportography, which shall only be employed if the indirect method of examination does not provide sufficient information, the patient has shunt capacity and an operation is to be performed immediately.


[Experimental investigations into dissolution of size of detail and contrast in the angiotomograms].

A comparative investigation on the resolution of angio-tomography and cut-film changer technique, standard views and direct geometrical radiographic magnification were carried out, using a phantom. Catheters of varying calibre and contrast were tested. The most significant result was the finding that lower concentrations of contrast were necessary to demonstrate small vessels or catheters when using angio-tomography or magnification, than with conventional techniques. It was also shown that angio-tomography and a magnification technique are able to show smaller and less contrasty vessels.


[New bases and theories from improving angio-cine-densitometry. II Improvements in the signal-to-noise ratio by the quotient method in cine densitometry].

In radio-cine-densitometry, random variations can be smoothed and reduced in two ways: temporal (frame to frame) and spatial (each frame). Both methods may, however, alter the signal measurements. The advantage of quotient cine-densitometry depends on the elimination of interference or random variations without changing the signal. Elimination of the variations occurs during signal recording. Amongst others the following are eliminated: variations in mains voltage, uneven development, absorption by other tissues and organs. A more suitable signal-to-noise ratio can be obtained. If an artery or a vein are opacified simultaneously it is possible, by means of quotient cine densitometry, to distinguish the arterial and venous phase and to analyse these separately from the overlying tissues.


Conformational differences in myosin, IV.[1-3] Radioactive labeling of specific thiol groups as influenced by ligand binding.

Changes in the mono- and divalentcation-stimulated ATPase activities of myosin progressively labeled with N-ethyl-[2,3-14C2]-maleimide were used to classify the readily reacting thiol groups into 3 types. The results show that one thiol-1 and one thiol-2 group are associated with each of the 2 active sites of myosin. Concentrations of KCl higher than 0.4M and/or temperatures above 10 degrees C lead to exposure of a variable number of thiol groups of a third class not affecting the enzymic properties. Although modification of thiol groups itself results in changes in structure and function of the protein, the patterns of incorporation of N-ethyl-[14C2]-malemide under various conditions of temperature, ionic strength and ligands bound to the protein revealed 9 different conformations of intact myosin. These were distinguished on the basis of the relative reactivity of the 3 different classes of thiol groups. The sequence of blockage of thiol groups reveals that cooperativity between the 2 active sites is induced by binding of a magnesium nucleotide complex to the protein. In the conformation of the long-lived myosin-product intermediate occuring during hydrolysis of Mg-ATP at 25 degrees C, 4 thiol groups of the third class react as well as or even more readily than those of the first and second classes.


Metabolism of sulfated glycosaminoglycans in cultivated bovine arterial cells. I. Characterization of different pools of sulfated glycosaminoglycans.

"Fibroblast-like" cells from the intimal layer of bovine aorta were grown in culture. The formation, composition, molecular weight and turnover rate of different pools of glycosaminoglycans were investigated in cultures incubated in the presence [35S]sulfate or [14C]glucosamine. The newly synthesized glycosaminoglycans are distributed into an extracellular pool (37 - 58%), a cell-membrane associated or pericellular pool (23 - 33%), and an intracellular pool (19 - 30%), each pool exhibiting a characteristic distribution pattern of chondroitin sulfate, dermatan sulfate, heparan sulfate and hyaluronate. The distribution pattern of the extracellular glycosaminoglycans resembles closely that found in bovine aorta. A small subfraction of the pericellular pool - tentatively named "undercellular" pool--has been characterized by its high heparan sulfate content. The intracellular and pericellular [35S]glycosaminoglycan pools reach a constant radioactivity after 8-12 h and 24 h, respectively, whereas the extracellular [35S]glycosaminoglycans are secreted into the medium at a linear rate over a period of at least 6 days. The intracellular glycosaminoglycans are mainly in the process of degradation, as indicated by their low molecular weight and by their half-life of 7 h, but intracellular dermatan sulfate is degraded more rapidly (half-life 4-5 h) than intracellular chondroitin sulfate and heparan sulfate (half-life 7-8 h). Glycosaminoglycans leave the pericellular pool with a half-life of 12-14 h by 2 different routes: about 60% disappear as macromolecules into the culture medium, and the remainder is pinocytosed and degraded to a large extent. Extracellular and at least a part of the pericellular glycosaminoglycans are proteoglycans. Even under dissociative conditions (4M guanidinium chloride) their hydrodynamic volume is sufficient for partial exclusion from Sepharose 4B gel. The existence of topographically distinct glycosaminoglycan pools with varying metabolic characteristics and differing accessibility for degradation requiresa reconsideration and a more reserved interpretation of results concerning the turnover rates of glycosaminoglycans as determined in arterial tissue.


Metabolism of sulfated glycosaminoglycans in cultivated bovine arterial cells. II. Quantitative studies on the uptake of 35SO4-labeled proteoglycans.

Cultured arterial fibroblasts were used for a quantitative study on adsorption, uptake and degradation of [35S]proteoglycans derived from secretions of cultured arterial or skin fibroblasts. The following results were obtained: 1) Proteoglycans added to the culture medium are integrated into the pool of cell membrane-associated (trypsin-removable) glycosaminoglycans by a saturable process, which depends on time and temperature. 2) Up to 17% of the added proteoglycans are taken up by the cells within 24 h. The uptake exhibits saturation kinetics, characteristic for adsorptive pinocytosis. Proteoglycan concentrations required for half-maximum uptake are higher than for half-maximum saturation of the glycosaminoglycan pool associated with the cell membrane. 3) After a lag phase, inorganic 35SO4 appears in the culture medium as a degradation product of the internalized proteoglycans. Pinocytosed proteoglycans are catabolized more rapidly than proteoglycans which remain inside the cell after their biosynthesis. 4) Pinocytosis exhibits specificity, the individual proteoglycans being internalized at different rates. The highest rate of uptake was measured for a dermatan-sulfate-rich proteoglycan. No competition of uptake between a dermatan-sulfate-rich and a heparan-sulfate-rich proteoglycan was observed. 5) Optimum pinocytosis requires an intact protein moiety and, presumably, undegraded carbohydrate chains of the proteoglycans.


Genetics of acheiropodia (the handless and footless families of Brazil). IX. Genetic counseling.

Acheiropodia offers no special problem of a counseling nature. The genetic risk can be easily estimated since the anomaly is due to a rare autosomal recessive gene with complete penetrance and little variation in expression. However, considering that acheiropod individuals have a tremendous handicap (they are born without both hands and feet), it is remarkable how they overcome this difficulty and lead an almost 'normal' life. Several examples are given in this paper. The 'philosophy' of genetic counseling is briefly discussed; emphasis is placed upon the necessity for the geneticist to present the good perspectives, while preparing the consultand for the worst. Even in this last possibility, however, not everything is necessarily lost as the example of the acheiropods attests.


[Clinical importance of hepatitis B antigen (author's transl)].

The clinical importance of the hepatitis B antigen (HBsAg) is based mainly on the differentiation between different courses of inflammatory liver diseases and/or nonvirus induced diseases of the liver. Epidemiologic studies have shown that beside a parenteral inoculation there is a possibility of a non-parenteral inoculation. Furthermore, there is evidence that the sporadic hepatitis is more commonly hepatitis B. Epidemiologic investigations revealed that clinically healthy HBsAg carriers, the "carrier" status in immunodeficiency syndromes and the natural circulation of the hepatitis B virus are of major interest. The demonstration of HBsAg enables us to characterize some diseases which show associations to hepatitis B virus, but where the pathogenic role of HBsAg/Ab immunocomplexes is questionable. Furthermore, it was possible by the detection of HBsAg to establish newer therapeutic and preventive interventions of virus B hepatitis.


Calf articular cartilage in organ culture in a chemically defined medium. 2. Concentrations of glycosaminoglycans and [35S]-sulfate incorporation at different oxygen tensions.

Articular cartilage from 6-month-old calves was maintained in organ culture in Eagle's minimum essential medium at different oxygen tensions between 20 and 50%. When well standardized cartilage pieces of about 1 mm thickness were used the results showed a high degree of reproducibility. The glycosaminoglycans were studied using the cetylpyridinium chloride cellulose and epichlorohydrin triethanolamine (ECTEOLA) column techniques on a microscale. In some experiments the cartilage was labelled with [35S]sulfate. Small alterations of the concentrations and distribution of different glycosaminoglycans were found and were not affected by various oxygen tensions for up to 4 weeks. A small increase was found in the fractions containing low molecular weight chondroitin sulfate. Devitalized cartilage, maintained under identical conditions, remained largely unaltered during the 4 weeks. The [35S]sulfate activity calculated per mole of hexosamine showed a general decrease during culture maintenance. High specific activities in the fractions containing low molecular weight chondroitin sulfate and keratan sulfate were found between 1 and 3 weeks of maintenance in 20% oxygen. Maintenance in 50% oxygen resulted in severely disturbed synthesis of glycoaminoglycans. The results are interpreted as showing that the synthesis pattern of glycosaminoglycans in articular cartilage can be altered by other factors and without the prior loss of glycosaminoglycans.


The inactivator of the first component of human complement (C1INA): the complex formation with plasmin.

The reaction of C1INA with plasmin was followed by the stoichiometric inactivation of both activities. On acrylamide gel electrophoresis, the reaction mixture revealed 3 new substances. One formed a precipitin band against anti-C1INA and its molecular weight was 103,000 daltons, 14,000 less than that of C1INA, indicating that a portion of C1INA was partially cleaved by the proteolytic activity of plasmin. Each of the other two substances formed precipitin bands against anti-C1INA as well as against anti-plasminogen. Molecular weights of these two substances were 200,000 and 179,000 daltons, whereas the molecular weights of C1INA and plasmin are 117,000 and 82,000 daltons, respectively. From these results, it was concluded that a portion of C1INA in the reaction mixture was partially cleaved by plasmin, and the partially cleaved C1INA as well as the native C1INA form 1:1 molecular complexes with plasmin, leading to inactivation of these activities.


The effect of vitamin E deficiency on androgen and corticosterone synthesis.

The incorporation of 14C-cholesterol into total androgens by testis homogenates and into total androgens and corticosterone by adrenal homogenates was measured in tissues taken from rats deficient in vitamin E, and from rats given vitamin E, from weaning to a maximum of 293 days. The peak of incorporation of 14C into adrenal steroids, androgens and corticosterone, were delayed in the rats deficient in vitamin E, which might suggest a delayed or prolonged period of puberty in vitamin E deficiency. After this period of increased steroidogenesis the deficient rats incorporated consistently less 14C into adrenal steroids and testicular androgens than did the control rats, although the differences were not statistically significant in the experiments described.


Hospitals harbor hazards ignored in fight for life.

Conventional images of the hospital create two two misconceptions in the minds of people having sporadic or casual contact with it: First, that it's not really a workplace, like a factory; and second that since the function of the hospital is to promote health and wellbeing, it's a healthy and safe place for those in it, whatever their capacity there. Both are far from the truth, as hospital administrators and the medical and operational staffs are beginning to realize and discuss with increasing intensity. Government agenicies at the state and federal level also are turning their scrutiny to the country's 7,000-plus hospitals and even-larger number of private and semi-private clinics. They employ several million people, and more millions are cared for-plus visitors and vendors who every day enter the hospital's doors, walk its corridors and are exposed to a variety of environmental conditions. In short, occupational health and safety is coming to the hospital. The hospital has many of the hazards of any workplace, such as electrical deficiencies in equipment, toxic gases in the atmosphere, noise and weights to lift. But it has countless more hazards that are unique to it: bacterial infection, ionizing and non-ionizing radiation, 24-hour-a-day, seven-days-a-week work schedules, and the fight for life in an intensive-care unit.


[Treatment of acne vulgaris using vitamin A acid].

Vitamin A acid represents the most effective therapeutic agent available for the topical treatment of acne vulgaris. This is also borne out by the results obtained in 152 patients over a two-year period. Clinical improvement is striking, long lasting remissions can be maintained by continuous treatment, and there appears to be an acceleration of the natural course to spontaneous remission of disease activity. Patients beyond their teens are more likely to remain symptom-free after adequate treatment than younger patients who may require prolonged maintenance treatment. Special attention is given to the appropriate care for Vitamin A acid treated skin and the general management of these patients is outlined. It is stressed that the patient be instructed, in detail, about the course of treatment and the necessity of staying under observation by an experienced dermatologist.


[The bullous variant of erythema annulare centrifugum Darier in Candida albicans infections].

A 15-year old male patient with Down-Syndrome suffering from a unusual vesicular type of erythema annular centrifugum Darier and a concurrent intestinal infection with candida albicans is described. Histology, immunhistological assay, and clinical observation allowed to differentiate Erythema annulare centrifugum from erythema exsudativum multiforme as well as dermatitis herpetiformis. On the basis of immunological findings and the clinical course, a hypersensitivity to candida albicans like a so-called "id-reaction" is assumed to be the main etiological factor in this case.


[Localization of solitary dermatophytic lesions in man].

The occurrence of three causative organisms of the so called intertriginous dermatophytoses--Epidermophyton floccosum, Trichophyton mentagrophytes var. interdigitale and T. rubrum--was compared according to the localization of the lesions. The differentiation of invasion into solitary and combined sites showed a prevalence of E. floccosum in solitary lesions and T. rubrum in combined lesions. In T. mentagrophytes var. interdigitale, the frequency of occurrence of both types of the lesions was approximately the same. The differences found suggest various adaptation-ability of dermatophytes to different parts of the body. The study of localization of solitary lesions is important from the epidemiological point of view because it assists in revealing the sources of dermatophytic infection and the ways of transmission.


Inhibition of adenosine triphosphatase and carbonic anhydrase by contrast media.

In vitro inhibition of adenosine triphosphatase and carbonic anhydrase by several radiographic contrast media and other compounds was measured. The concentrations required for 50% inhibition were, in general, similar to those obtained for other enzymes. The effect of methylglucamine could not be determined for either enzyme because of interference with the assays.


[Inhibition and stimulation of ATPase activity and the nonenzymatic hydrolysis of ATP in electron histochemistry (author's transl)].

In rat liver, homogenized and fractionated, ATPase activity was studied both histochemically and biochemically in the presence of lead ions and with addition of cysteine, EDTA, 2,4-DNP, sodium cyanide, sodium fluoride and p-chloromercuribenzoate. None of the inhibitors concerned permitted to obtain a complete inhibition of ATP hydrolysis. A practically complete inhibition of ATPase was possible only through heat inactivation or after double prefixation with formaldehyde and ethanol. Nonenzymatic ATP hydrolysis plays a negligible role in ATPase histochemistry since phosphate yield taking place due to lead action is insignificant against enzymatic hydrolysis of ATP.


An ultrastructural study of neuromuscular spindles in normal mice: with reference to mice and man infected with Mycobacterium leprae.

Mycobacterium leprae have been found within muscle spindles in mice, using electron microscopy, and in man, using light microscopy. Their mode of entry clearly is important. It may be via capsular cells, capillaries or nerves. For this reason muscle spindles from normal mice were studied by electron microscopy with special reference to the capsule and the relationship of it with capillaries and nerves, as well as details of the intrafusal fibres and capsular space. A fenestrated capillary was found between the capsular cell layers and outside the capsule in one spindle and in another spindle both a fenestrated and a continuous type of a capillary were found between the capsular cells; this may be of interest for pharmacological studies. However, the muscle spindles of the mouse were in the main similar to muscle spindles studied by other workers in man and other mammals.


The disturbance of oxidative phosphorylation by N-acetoxy-N-acetyl-2-aminofluorene, a model ultimate carcinogen.

Currently N-acetoxy-N-acetyl-2-aminofluorene is favored by many investigators to be a model of the ultimate electrophilic carcinogenic agent derived metabolically from the carcinogen N-acetyl-2-aminofluprene. The model induced in vitro a delayed ATP energized increase in mitochondrial volume as indicated by the decrease in absorbancy at 520 nm. The ATP energized decrease in absorbancy was inhibited by rutamycin, 2,4-dinitrophenol and a high level of antimycin known to induce ATPase activity. The known to inhibit respiration without inducing ATPase activity. Malate or potassium ion did not affect the phenomenon, however, sulfate ion which has been implicated in liver carcinogenesis shortened the induction period. Showdomycin stimulated the phenomenon. N-Acetoxy-N-acetyl-2-aminofluorene interacts with the machinery of oxidative phosphorylation. N-Acetoxy-N-acetyl-2-aminofluorene was enzymically converted by the mitochondria to N-hydroxy-N-acetyl-2-aminofluorene. These findings extend the experimental confluence of oxidative phosphorylation with carcinogenesis.


Design of living environments for nursing-home residents: increasing participation in recreation activities.

Nursing-home residents have frequently been characterized as unoccupied and disengaged. At the outset of the present study, most residents were to be found in their own rooms, not exhibiting gross motor behavior or social interaction, and not participating in appropriate activities. To modify residents' levels of participation with the environment, a manipulative area was provided in the lounge. Participation in the lounge averaged 20% on days when the activity was not available, but increased to a mean of 74% on days when equipment and materials were given and residents were prompted to participate. When prompts were withdrawn and materials were available only by request, mean participation fell to 25%. The findings demonstrate that manipulative activities can support a high level of participation with the environment, if residents are prompted to use equipment and materials.


Relationship between the tonB locus and iron transport in Escherichia coli.

When a strain (arcB-) of Escherichia coli, unable to synthesize the iron transport compound enterochelin, was transduced to tonB-, it became resistant to phage phi80 and simultaneously lost the growth response to enterochelin and the ability to transport its iron complex. However, enterochelin precursors (shikimate and 2,3-dihydroxybenzoate) still supported growth, via the synthesis of enterochelin. Dihydroxybenzoate was a better growth factor at a low concentration than it was at higher levels. The evidence suggests that tonB- strains lack an outer membrane component necessary both for the uptake of ferric-enterochelin and for the adsorption of phage phi80. Thus, although ferric-enterochelin cannot penetrate the cell surface from outside, the complex that is formed within the envelope is transported normally into the cell. The aroB-, tonB- mutant also lacked growth reponses to citrate and various hydroxamate siderochromes, which supported growth in the tonB+ parent strain via inducible transport systems for their ferric complexes. The aroB-, tonB- mutant was unable to transport iron in the presence of citrate, but the low-affinity uptake of uncomplexed iron and the transport of amino acids and phosphate were unimpaired. The tonB locus, thus, affects all the known active transport systems for iron, possibly indicating that they share some common outer membrane component.


Bacteriophage-typable revertants from pleiotropic staphylococcal mutants.

Cell walls were physically purified from bacteriophage-typable revertants that had been isolated from modified cell wall pleiotropic strains derived from Staphylococcus aureus NCTC 8511. The quantitative amino acid, amino sugar, and phosphorus contents of these cell walls are reported. Among the revertants were some whose walls possessed elevated serine and one strain whose walls contained the novel amino sugar galactosamine. The similarities in bacteriophage typing patterns between the revertants and the original parental strain lead to the conclusion that the previously described pleiotropic strains are mutants of NCTC 8511.


Characterization of 5.8S ribosomal ribonucleic acid in Neurospora crassa.

Neurospora crassa ribosomes contain a species of ribonucleic acid (RNA) of molecular weight 54,000, similar to 5.8S ribosomal RNA previously described for other eukaryotic organisms. The 5.8S RNA from N. crassa was found to be released by heat treatment at 60 C from 25S ribosomal RNA but not from 18S ribosomal RNA. The base composition of N. crassa 5.8S RNA was similar to that of 5.8S RNA from Saccharomyces cerevisiae, but differed from animal 5.8S RNA. During the course of this study, it was discovered that N. crassa 25S ribosomal RNA had a number of internal cleavages that may exist in vivo.


Siroheme: a prosthetic group of the Neurospora crassa assimilatory nitrite reductase.

The Neurospora crassa assimilatory nitrite reductase (EC 1.6.6.4) catalyzes the NADPH-dependent reduction of nitrite to ammonia, a 6-electron transfer reaction. Highly purified preparations of this enzyme exhibit absorption spectra which suggest the presence of a heme component (wavelength maxima for oxidized senzyme: 390 and 578 nm). There is a close correspondence between nitrite reductase activity and absorbance at 400 nm when partially purified nitrite reductase preparations are subjected to sucrose gradient centrifugation. In addition, a role for an iron component in the formation of active nitrite reductase is indicated by the fact that nitrate-induced production of nitrite reductase activity in Neurospora mycelia in vivo requires the presence of iron in the induction medium. The heme chromophore present in Neurospora nitrite reductase preparations is reducible by NADPH. Complete reduction, however, requires the presence of added FAD. The NADPH-nitrite reductase activity of the enzyme is also dependent upon addition of FAD. A spectrally unique complex is formed between the heme chromophore and nitrite (or a reduction product thereof) when nitrite is added to NADPH-reducted enzyme. Carbon monoxide forms a complex with the heme chromophore of nitrite reductase with an intense alpha-band maximum at 590 nm and a beta-band of lower intensity at 550 nm. CO is an inhibitor of NADPH-nitrite reductase activity. Spectrophotometrically detectable CO complex formation and Co inhibition of enzyme activity share the following properties...


Identification of the dicyclohexylcarbodiimide-reactive protein component of the adenosine 5'-triphosphate energy-transducing system of Escherichia coli.

Membranes of Escherichia coli contain an adenosine 5'-triphosphate (ATP) energy-transducing system that is inhibited by treatment with dicyclohexylcarbodiimide (DCCD). The carbodiimide-reactive protein component of this system has been identified after treatment with [14C]DCCD. This protein has an apparent molecular weight of 9,000 as judged from acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and is extracted from the membrane with chloroform-methanol (2:1). These properties are similar to the analogous protein previously identified in mitochondria (Cattell et al., 1971). A mutant strain, RF-7, has been isolated which derives energy from oxidative phosphorylation in the presence of 5 mM DCCD. The ATP hydrolase activity of the membraned system in the mutant was considerably less sensitive to inhibition by DCCD than that in the wild type. The carbodiimide-reactive protein, which was easily labeled by [14C]DCCD in the wild type, was labeled much less rapidly in the carbodiimide-resistant mutant. It is thus concluded that the reaction of DCCD with this specific protein leads to inhibition of the ATP energy-transducing reactions. The mutation causing carbodiimide resistance in strain RF-7 was mapped. It is cotransduced with the uncA gene at a frequency exceeding 90%. The mutationally altered protein causing the carbodiimide resistance was not conclusively identified. However, reconstitution experiments indicate that the altered protein is not one of the subunits of the soluble ATP hydrolase activity, which can be removed from the membrane by washing with 1 mM tris(hydroxymethyl)aminomethane buffer lacking Mg2+. The carbodiimide-reactive protein remains with the membrane residue after removal of the soluble ATP hydrolase and is thus distinct from these subunits as well.


Effects of urinary proteins from certain leukemics upon macromolecular synthesis and enzyme levels in bone marrow cultures.

Urinary proteins from human leukemic patients have been found to alter quantitatively macromolecular synthesis in primary mouse bone marrow cultures. Urinary protein-stimulated incorporation of [3H]uridine into RNA was found after 1 day of culture. Increased levels of adenine phosphoribosyltransferase and lysozyme were demonstrable at 3 and 5 days, respectively, with urinary protein-supplemented cultures. The incorporation of 3H-labeled deoxynucleosides into DNA was higher in the presence of urinary proteins after 2 days of culture. The rate of incorporation of [3H]deoxyuridine into DNA was strongly inhibited by 10(-5) M Methotrexate and 10(-6) M 5-fluorodeoxyuridine, however, the effect of urinary proteins on incorporation of [3H]uridine into RNA and lysozyme accumulation were not inhibited. Urinary proteins also stimulated the formation of "colonies" (groups of at least 30 cells) in media containing methylcellulose. This latter phenomenon was also not inhibited by 10(-5) M Methotrexate or 10(-6) M 5-fluorodeoxyuridine. The results of these studies are consistent with the postulate that in the presence of human urinary proteins, mouse bone marrow cells in culture proceed to a phenotype characteristic of circulating peripheral white cells.


High-resolution biomedical gas chromatography. Determination of human urinary steroid metabolites using glass open tubular capillary columns.

Recently developed analytical procedures for the qualitative and quantitative analysis of human urine for major and minor steroid metabolites are described. Steroid profile samples were obtained by enzymic hydrolysis with beta-glucuronidase  and sulfatase. Methoxime-trimethylsilyl ether derivatives of steroid metabolites were prepared; the recommended procedure converts all ketone groups (except the 11-one group) into methoxime groups and all hydroxyl groups into trimethylsily ether groups. These derivatives are thermally stable, readily volatilized, not subject to dehydration or adsorption on gas chromatographic columns, and suitable for both quanlitative and quantitative analytical studies. Thermostable glass open tubular capillary columns, coated with the non-polar phase SE-30, and containing dispersed particles of silanized silicic acid, were used for the gas chromatographic separation. Illustrations of profiles for normal female and male subjects, and patients with a testosterone-secreting ovarian tumor, congenital adrenal insufficiency and a dehydroepiandrosterone-secreting adrenal tumor are included.


Studies on steroids. III. A new type of derivative for electron capture-gas chromatography of ketosteroids.

Pentafluorobenzyloxyamine is presented as a new derivatizing agent for gas chromatography of ketones using electron capture detection. The typical 17-ketosteroid was readily transformed into the O-pentafluorobenzyloxime which on usual trimethylsilylation led to a 3-trimethylsilyl ether derivative exhibiting good gas chromatographic properties. The derivatization procedure was applied to the determination of dehydroepiandrosterone in human plasma by electron capture-gas chromatography employing an internal standard method and in consequence satisfactory results were obtained.


Treatment of aortic arch aneurysms with deep hypothermia and circulatory arrest.

A patient with an aneurysm of the aortic arch is presented. The patient underwent successful surgical correction using deep hypothermia with circulatory arrest and preserving an aortic cuff including the supra-aortic trunks to facilitate the anastomosis with the prosthesis. Technical and tactical detail are also discussed.