[Comparative characteristics of arteriosclerosis of the aorta, subclavian, iliac and femoral arteries in the populations of Riga and Samarkand]. Visual-planimetric evaluation of atherosclerosis of the aorta, subclavian, iliac, and femoral arteries in men of 50--64 who had died in Riga and Samarkand was carried out. The values characterizing the extent and severity of atherosclerosis of these arteries in the indigenous and non-indigenous population of these cities were calculated. Differences in the morphological features and course of atherosclerosis of arteries of the extremities in relation to the place of residence were established. No significant differences in the development of atherosclerotic changes of arteries of the extremities between indigenous and non-indigenous residents of Riga were found. The indigenous residents of Samarkand had low values of the area of atherosclerotic changes of the blood vessels of the extremities as well as low frequency of stenoses and obliterating lesions as compared with the non-indigenous residents. The observed differences indicate the importance of climatic-geographical and genetic factors and national-environmental features of the way of life in the pathogenesis of atherosclerotic diseases of the extremities.


[Tissue-specific regulation of proliferative processes in the tissues normally and in pathology: on the problem of chalones]. The paper discusses the author's own and literary data on the role of chalones (tissue-specific inhibitors of proliferative processes) in the maintenance of tissue homeostasis normally and pathologically. It is suggested that chalones be used both as regulators of cell multiplication and as markers of histogenesis.


The functional unit of calcium-plus-magnesium-ion-dependent adenosine triphosphatase from sarcoplasmic reticulum. The aggregational state of the deoxycholate-solubilized protein in an enzymically active form. Vesicles consisting of (Ca(2+)+Mg(2+))-dependent ATPase (adenosine triphosphatase), and lipid were prepared from sarcoplasmic reticulum of rabbit skeletal muscle. As with non-ionic detergents [le Maire, Møller & Tanford (1976) Biochemistry15, 2336-2342] the (Ca(2+)+Mg(2+))-dependent ATPase after solubilization by deoxycholate showed a pronounced tendency to form oligomers in gel-chromatographic experiments, when eluted in the presence of deoxycholate and phosphatidylcholine. To evaluate the functional significance of oligomer formation the properties of enzymically active preparations of ATPase, solubilized by deoxycholate, were studied. Such preparations were obtained at a protein concentration of 2.5mg/ml in the presence of a high salt concentration (0.4m-KCl) and sucrose (0.3m) in the solubilization medium. Analytical ultracentrifugation of solubilized ATPase showed one protein boundary moving at the same rate as gel-chromatographically prepared monomeric ATPase (s(20,w)=6.0S). From simultaneous measurements of the diffusion coefficient an apparent molecular weight of 133000 was calculated, consistent with solubilization of ATPase in predominantly monomeric form. The enzymic activity of deoxycholate-solubilized ATPase when measured directly in the solubilization medium at optimal Ca(2+) and MgATP concentrations was about 35-50% of that of vesicular ATPase. The dependence of enzymic activity on MgATP concentration indicated that the solubilized ATPase retained high-affinity binding of MgATP, but the presence of high concentrations of the nucleotide did not stimulate activity further, in contrast with that of vesicular ATPase. The dependence of enzymic activity on the free Ca(2+) concentration was essentially the same for both solubilized and vesicular forms, indicating that interaction of ATPase with more than one molecule of Ca(2+) is required for enzyme activity. Solubilized enzyme at 20 degrees C was phosphorylated to about the same degree as vesicular ATPase. It is concluded that the catalytic activity of monomeric ATPase retains most of the features of vesicular ATPase and that extensive oligomer formation in gel-chromatographic experiments in the presence of deoxycholate probably reflects processes taking place during inactivation and delipidation of the protein.


Gliding edge dislocations in proteins as a mechanism for active ion transport. A mechanism for the translocation of ions across biological membranes is proposed in which transconformational changes occur in the membrane-bound ion pump. In particular, it is suggested that such conformational changes are effected by means of the propagation of crystal defects in the form of edge-dislocation glide.


Comparison of alpha-methylphenylalanine and p-chlorophenylalanine as inducers of chronic hyperphenylalaninaemia in developing rats. alpha-Methylphenylalanine is a very weak competitive inhibitor of rat liver phenylalanine hydroxylase in vitro but a potent suppressor in vivo. The loss of the hepatic activity (the renal one is unaffected) becomes maximal (70-75% decrease; cf. control) 18h after the administration (per 10g body wt.) of 24 mumol of alpha-methylphenylalanine with or without 52 mumol of phenylalanine. Chronic suppression of hepatic phenylalanine hydroxylase was obtained by injections of alpha-methylphenylalanine plus phenylalanine to suckling rats, and by their addition to the diet after weaning. A series of comparisons of the effects of this treatment, and one with p-chlorophenylalanine, was then carried out. In both cases there was a rise (1.3-2-fold) in phenylalanine-pyruvate amino-transferase activity (but no change in four other enzyme activities) in the liver; in brain there was a rise in phosphoserine phosphatase activity, but the total activity and subcellular distribution of nine enzymes revealed no other abnormalities in cerebral development. Striking increases in the concentration of plasma phenylalanine during 26 of the 31 experimental days (with a transient fall at 18-22 days) were maintained by treatment with both analogues plus phenylalanine. However, p-chlorophenylalanine-treated animals had a 30-60% mortality rate and 27-52% decrease in body weight. Developing rats treated with alpha-methylphenylalanine, showing no growth deficit or signs of toxicity (e.g. cataracts), appear to be a more suitable model for the human disease of phenylketonuria. Their phenylalanine concentrations exhibited at least 20-40-fold increase during 50% of each of the first 18 days of life, and 30-fold after weaning.


Citreoviridin, a specific inhibitor of the mitochondiral adenosine triphosphatase. 1. Citreoviridin was a potent inhibitor of the soluble mitochondrial ATPase (adenosine triphosphatase) similar to the closely related aurovertins B and D. 2. Citreoviridin inhibited the following mitochondrial energy-linked reactions also: ADP-stimulated respiration in whole mitochondria from ox heart and rat liver; ATP-driven reduction of NAD+ by succinate; ATP-driven NAD transhydrogenase and ATPase from ox heart submitochondrial particles. 3. The dissociation constant (KD) calculated by a simple law-of-mass-action treatment for the citreoviridin--ATPase complex was 0.5--4.2micron for ox-heart mitochondrial preparations and 0.15micron for rat liver mitochondria. 4. Monoacetylation of citreoviridin decreased its inhibitory potency (KD=2--25micron, ox heart; KD=0.7micron, rat liver). Diacetylation greatly decreased the inhibitory potency (KD=60--215micron, ox heart). 5. Hydrogenation of citreoviridin monoacetate diminished its inhibitory potency considerably. 6. No significant enhancement of fluorescence was observed when citreoviridin interacted with the mitochondrial ATPase.


Genetic complementation between two mutant unc alleles (unc A401 and unc D409) affecting the Fl portion of the magnesium ion-stimulated adenosine triphosphatase of Escherichia coli K12. A new mutant strain of Escherichia coli in which phosphorylation is uncoupled from electron transport was isolated. A genetic-complementation analysis, using partial diploid strains, showed that the new mutant allele, uncD409, is in a gene distinct from the other previously identified genes uncA, uncB and uncC. A strain carrying the uncd409 allele has no Mg2+ ion-stimulated adenosine triphosphatase activity and is therefore phenotypically similar to strains carrying the uncA401 mutant allele. Complementation between the uncA401 and the uncD409 alleles occurred, as indicated by growth of partial diploid strains on succinate and their growth yields on limiting concentrations of glucose. Complementation was confirmed by using membranes prepared from the above partial diploids. Such membranes were found to have Mg2+-stimulated adenosine triphosphatase activity, ATP-dependent transhydrogenase activity ADP-induced atebrin-fluorescence quenching and low but significant amounts of oxidative phosphorylation.


Preparation and characterization of frog muscle myosin subfragment 1 and actin. The preparation, structural and steady-state kinetic characteristics of contractile proteins from the leg muscle of frogs Rana temporaria and Rana pipiens are described. Actin and myosin from the two frog species are indistinguishable. The proteins have structural and steady-state kinetic properties similar to those from rabbit fast-twitch skeletal muscle. Chymotrypsin digestion of frog myosin or myofibrils in the presence of EDTA yields subfragment 1, which is separated by chromatography into two components that are distinguished by their alkali light-chain content.


Reaction mechanism of the magnesium ion-dependent adenosine triphosphatase of frog muscle myosin and subfragment 1. The Mg2+-dependent ATPase (adenosine 5'-triphosphatase) mechanism of myosin and subfragment 1 prepared from frog leg muscle was investigated by transient kinetic technique. The results show that in general terms the mechanism is similar to that of the rabbit skeletal-muscle myosin ATPase. During subfragment-1 ATPase activity at 0-5 degrees C pH 7.0 and I0.15, the predominant component of the steady-state intermediate is a subfragment-1-products complex (E.ADP.Pi). Binary subfragment-1-ATP (E.ATP) and subfragment-1-ADP (E.ADP) complexes are the other main components of the steady-state intermediate, the relative concentrations of the three components E.ATP, E.ADP.Pi and E.ADP being 5.5:92.5:2.0 respectively. The frog myosin ATPase mechanism is distinguished from that of the rabbit at 0-5 degrees C by the low steady-state concentrations of E.ATP and E.ADP relative to that of E.ADP.Pi and can be described by: E + ATP k' + 1 in equilibrium k' - 1 E.ATP k' + 2 in equilibrium k' - 2 E.ADP.Pi k' + 3 in equilibrium k' - 3 E.ADP + Pi k' + 4 in equilibrium k' - 4 E + ADP. In the above conditions successive forward rate constants have values: k' + 1, 1.1 X 10(5)M-1.S-1; k' + 2 greater than 5s-1; k' + 3, 0.011 s-1; k' + 4, 0.5 s-1; k'-1 is probably less than 0.006s-1. The observed second-order rate constants of the association of actin to subfragment 1 and of ATP-induced dissociation of the actin-subfragment-1 complex are 5.5 X 10(4) M-1.S-1 and 7.4 X 10(5) M-1.S-1 respectively at 2-5 degrees C and pH 7.0. The physiological implications of these results are discussed.


Heterogeneity in the composition and aggregation patterns of coronary intima acid mucopolysaccharides (glycosaminoglycans). The thickened intima of human coronary arteries included acid mucopolysaccharides (glycosaminoglycans) with a large variety of patterns of composition and aggregation. This heterogeneity, detectable with histochemical methods, was related in our material to age, branching anatomical pattern, size of vessel, collateral or terminal character of the vessel, bifurcations, intimal histology, internal elastic membrane alterations, as well as to the onset of fatty streaks, gelatinous plaques and incorporated microthrombi.


Menstrual patterns after laparoscopic sterilization using a spring-loaded clip. Details of the menstrual cycles in 504 women were recorded before laparoscopic sterilization with a spring-loaded clip and at six months and one year after the procedure. The operation did not affect the length of the menstrual cycle or duration of menstrual flow, nor the patient's assessment of the amount of blood losss or the incidence of dysmenorrhoea. The previous method of contraception was significantly related to subsequent menstrual patterns.


Studies of (Na+ + K+)-sensitive ATPase activity in pig lymphocytes. Effects of concanavalin A. (Na+ + K+)-ATPase activity is demonstrated in plasma membranes from pig mesenteric lymph nodes. After dodecyl sulfate treatment plasma membranes have an 18-fold higher (Na+ + K+)-ATPase activity, while their ouabain-insensitive Mg2+-ATPase is markedly lowered. A solubilized (Na+ +K+)-ATPase fraction, obtained by Lubrol WX treatment of the membranes, has very high specific activity (21 mumol Pi/h per mg protein). Concanavalin A has no effect on these partially purified (Na+ + K+)-ATPase, while inhibits (40%) this activity in less purified fractions which still contain Mg2+-ATPase activity.


Reversible shift between two states of Ca2+-ATPase in human erythrocytes mediated by Ca2+ and a membrane-bound activator. The (Ca2+ + Mg2+)-dependent ATPase (ATP phosphohydrolase, EC 3.6.1.3) from human erythrocytes occurred in two different states, A-state and B-state, depending on the membrane preparation. The A-state showed low maximum activity (V) and the Ca2+ activation was characterized by a Hill coefficient, nH, of about 1 and a Michaelis constant, KCa, about 30 micron. The B-state showed high V, a nH above 1, which indicates positive cooperativity of Ca2+ activation, and KCa of about 1 micron. With varying ATP concentrations, both the A-state and B-state showed negative cooperativity and slightly different values of Km. The B-state was shifted to A-state when the membranes were exposed to low Ca2+ concentration. The shift reached 50% at approx. 0.5 micron Ca2+. At the low Ca2+ concentrations an activator was released from the membranes. The A-state was shifted to the B-state when the membranes were exposed to Ca2+ in the presence of the activator. The shift reached 50% at about 30 micron Ca2+. The recovery of high V was time dependent and lasted several minutes. Increasing concentrations of Ca2+ and activator accelerated the recovery. It is suggested that the A-state and the B-state correspond to enzyme free of activator and enzyme associated with activator, respectively. Furthermore, the two states may respresent a resting and an active state, respectively, of the calcium pump.


Dynamic light scattering characterization of the detergent-free, delipidated (Ca2+ + Mg2+)-ATPase from sarcoplasmic reticulum. Dynamic light scattering studies have been conducted on the delipidated and detergent-removed (Ca2+ + Mg2+)-ATPase protein assemblies. Specific characterization of the state of aggregation and the extent of conformation change upon delipidation and detergent removal has been made. The results show that the prominent species are dimers and tetramers of very globular nature, with axial ratios of less than 2 : 1. The hydrodynamic radii of the dimers and the tetramers are, respectively, 57.5 A and 74.5 A. The globular nature of these observed entities differ from the delipidated ATPase proteins recently obtained (LeMaire, M., Jorgensen, K.E., Roigaard-Petersen, H. and Moller, J.V. (1976) Biochemistry 15, 5805--5812. Present results suggest that upon the removal of detergents from the lipid-free ATPase protein assembly, only a rather limited degree of aggregation takes place. Such a condition is consistent with models of the membrane protein system which has limited regions of hydrophobic contact. Oligomeric assemblies with aqueous channels is a possible active Ca2+ transport model consistent with results of the present data, as well as the data from several other recent studies.


Reversible superprecipitation and bundle formation of plasmodium actomyosin. Synthetic actomyosin from plasmodium was found to undergo reversible superprecipitation upon addition of ATP. According to electronmicroscopic investigation upon clearing, short myosin filaments of about 0.2 micron in length appeared predominantly coexisting with actin filaments, and after superprecipitation, bundles of actin filaments were formed where short myosin filaments or myosin molecules were bound to the side of the bundle, making a whisk-like structure. The turbidity and the ATPase activity of actomyosin were measured at various ATP concentrations clamped by using an ATP-regenerating system. The turbidity was high below 1 . 10(-6) M ATP, corresponding to the state of superprecipitation, and with increasing ATP concentration it dropped in the range of 1 . 10(-6)--1 . 10(-5) M ATP. On the other hand, the ATPase activity was low below 1 . 10(-6) M ATP and increased above 1 . 10(-5) M after the turbidity dropped. Characteristic features of superprecipitation of plasmodium actomyosin observed here were discussed in relation to the mechanism of motility in vivo.


[Phase transitions of model and biological membranes. II. Temperature-dependent structural reorganization of sarcoplasmic reticulum membranes]. Temperature-dependent structural transitions of sarcoplasmic reticulum membranes, Ca2+-activated ATPase from membranes and liposomes formed from total membrane lipids were studied by fluorescent probes such as 3-metoxybenzantron and dimetylaminochalkon. It was found that sarcoplasmic reticulum membranes undergo the structural transitions at 10, 17--18, 30, 41--43 and 49 degrees C; lipids at 14--15, 30 degrees C and ATPase at 47--49 degrees C. The data obtained here indicate that transitions at 10, 17--18 and 30 degrees C in sarcoplasmic reticulum membranes are associated with a melt of lipids, while the transitions at 41--43 and 49 degrees C apparently reflect changes in conformation of membrane proteins.


[Thermodynamic muscle cycles].

[Thermodynamic muscle cycles]. On the basis of the analysis of muscle thermodynamic cycles in terms of the concept of non-equilibrium accumulator of energy some conclusions can be made concerning the character of temperature--entropy relationships. If the redistribution phase proceeds with an essential increase of entropy, the muscle contraction cycle proves to be more advantageous energetically at negative absolute temperatures. If the entropy in this phase gets considerably lower it is more advantageous to realize the cycle in the range of positive absolute temperatures. The non-linearity of ATPase activity which has the maximum in the intermediate phases of contraction leads to the "Maxwell quasidemon" effect and to an increase of overall efficiency.


[The effect of membrane-bound calcium on the activity of adenosine triphosphatase from erythrocytes and erythrocyte permeability for monovalent cations]. The activities of Ca2+, Mg2+-ATPase and Na+, K+-ATPase and the permeability of reconstituted human erythrocytes for Na and K ions were measured, using Ca2+-EGTA, Ca2+ATP and Ca2+-sodium citrate buffers. It was found that the increase in the Ca2+/chelate ratio caused stimulation of Ca2+, Mg2+- and Na+, K+-Atpases and an increase in the rate constants of ouabain--dependent 42K+ influx and 22Na+ efflux from the erythrocytes. The use of the Ca2+-sodium citrate system as a calcium buffer did not change the parameters of the functional state of erythrocyte membranes. The data obtained are discussed in terms of a possible role of calcium ions, which are bound to the inner surface of the erythrocyte membrane, in the regulation of the systems of active and passive transport of cations.


Study of a kindred with partial deficiency of red cell 2,3-diphosphoglycerate mutase (2,3-DPGM) and compensated hemolysis. A kindred with partial deficiency of red cell 2,3-diphosphoglycerate mutase (2,3-DPGM) was studied. The propositus presented with indirect hyperbilirubinemia, normal hemoglobin (15.8 g/dl), and elevated reticulocyte count (4.6%). The red cell 51Cr survival was decreased (tau1/2 16 days). Incubated osmotic fragility was normal; autohemolysis was increased and corrected with glucose and ATP. The P50 was 18.5 mm Hg (normal 25.5 +/- 3), but the stability, electrophoresis, and fingerprinting of hemoglobin were normal. The concentration of 2,3-diphosphoglycerate (2,3-DPG) was reduced to 43% of normal. Red cell 2,3-DPGM was decreased to 59% of normal; 2,3-DPG phosphatase was similarly decreased. All red cell glycolytic and hexose monophosphate shunt enzymes, glycolytic intermediates other than 2,3-DPG, and glucose consumption and lactate production were normal. Five family members showed similar hematologic findings. The deficiency appears to be secondary to decreased enzyme synthesis and to be inherited as an autosomal dominant trait in this family. Partial deficiency of 2,3-DPGM should now be considered in the differential diagnosis of compensated hemolysis associated with increased oxygen affinity.


Regional brain atrophy and reductions in glutamate release and uptake after intrastriatal kainic acid. 1 Neurochemical changes and tissue weights were measured following intrastriatal injection of 2.5 microgram of kainic acid in 2 microliter of 0.9% w/v NaCl solution (saline) in the rat. 2 After kainic acid the striatum and neocortex on the injected side showed a progressive reduction in weight, the neocortex showing the greatest absolute weight loss and the striatum the greatest percentage change. 3 Large (80-90%) reduction in choline acetyltransferase (CAT) and L-glutamate decarboxylase (GAD) activities in the striatum occurred within 2-4 days of the injection and persisted at least 10 weeks. At 10 weeks CAT and GAD activities were unaltered in the neocortex. 4 The absolute content of dopamine in the striatum was not different from control 5 days after the injection of kainic acid but was reduced at 2 and 10 weeks. At 2 weeks the concentration (microgram/g wet weight) of dopamine also was reduced but at 10 weeks it was near normal due to atrophy of the striatum. 5 The high affinity glutamate uptake into a crude synaptosomal preparation of the striatum was reduced by 64% 5 days after kainic acid and still reduced by 67% at 10 weeks. 6 The efflux of glutamate from slices of the striatum in the presence of 52 mM K+ was reduced by approximately 75% 5 days and 15 weeks after kainic acid. 7 In vitro kainic acid (10(-4) M) neither altered the high affinity uptake of radiolabelled glutamate into a homogenate of the striatum, nor released endogenous glutamate from slices of striatum.


Whorl-like degeneration of corneal epithelium a clinico-pathologic study. A case of epithelial corneal degeneration that was first seen as a pigmented plaque but changed to a whrol-like pattern and then partially regressed is described. The pathologic abnormality was an intra-cellular inclusion of lipid in the basal cells and of lipid and mucopolysaccharide in the wing and superficial cells. No duplication of basement membrane or thickening of Bowman's layer was present. It is suggested that despite different origins the pathologic entity in this case, in Fabry's disease and in chloroquine keratopathy is similar. The whorled pattern is due to a sliding of the corneal epithelial cells as they rise from the basal cells to the surface, much as a cyclonic flow forms about an atmospheric low in the southern hemisphere.


An experimental model of acute immune-complex uveitis. We developed an experimental model of anterior uveitis by injecting ovalbumin or bovine serum albumin into the anterior chamber of sensitized rabbits. All the eyes showed leakage of serum proteins into the aqueous humor within 30 min. and developed uveitis within 24 hrs. which lasted at least a week. On histological examination, large numbers of polymorphonuclear (PMN) leucocytes and only occasional mononuclear cells were seen in the iris, the ciliary body and the irido-corneal angle. Immunohistological examination revealed 7s gamma globulin in the walls of blood vessels, in the interstitium and inside the PMN leucocytes. Contact radiographs of eyes challenged repeatedly showed extensive neovascularization of the iris and ciliary body.


Effects of ethylene on the metabolism of Saccharomyces cerevisiae. Supply of exogenous ethylene to lactate-grown yeast initially accelerated the rate of ethanol production from glucose, but later reduced the rate, with the overall effect being to reduce the total ethanol production. The rate of ethanol production by ethylene-treated yeast was not changed by removal of metabolic carbon dioxide. However, if CO2 was allowed to build up in the absence of applied ethylene, the ethanol production decreased. Ethylene increased the activities of a number of pentose phosphate and glycolytic pathway enzymes. The largest increase in activity was observed for phosphofructokinase (EC 2.7.1.11), regulatory enzyme of the glycolytic pathway. After an initial stimulation, glucose (and also 3-O-methyl glucose) uptake was reduced by ethylene. Ethylene appears to inhibit non-competitively the glucose transport system.


Miniature end-plate potentials in the frog muscle spindle. Spontaneous miniature end-plate potentials (m.e.p.p.s) were recorded from the intrafusal muscle fibers in the muscle spindles isolated from the frog sartorius and semitendinosus muscles by means of microelectrode (intracellularly) and across a vaseline gap made transversely on the intrafusal bundle (extracellularly). The latter method was available to record simultaneously the m.e.p.p.s and the afferent terminal activities, of which the latter was easily distinguishable from the former by concomitant occurrence with the axonal responses. Simple type spindles in the sartorius muscle have a restricted end-plate region along the intrafusal bundle on either side of the capsule, while complex type spindles in the semitendinosus muscle are innervated by plural regions which are found on both sides of the capsule in most preparations. Application of 2X hypertonic Ringer's solutions with NaCl or sucrose induced an increase in the rate of the m.e.p.p.s preceding a decrease in the rate of afferent discharges from the spindle.


Emerging treatments: replacement therapy with choline or lecithin in neurological diseases. This review evaluates the theoretical background and experimental data behind a new development: the replacement therapy of deficient central cholinergic systems with the dietary precursors choline or lecithin. Cholinergic deficiency states are possibly present in five neurological entities: Huntington's chorea, Tardive Dyskinesia, Gilles de la Tourette's disease, Friedreich's ataxia and pre-senile dementia. Preliminary data from various laboratories, including our own, in each of these disorders indicate that some clinical improvement can occasionally be seen, and that this approach deserves  further investigation.


Streptozotocin in advanced small cell bronchogenic carcinoma: an ineffective nonmyelosuppressive agent. Thirteen patients with advanced small cell bronchogenic carcinoma (SCBC) received at least three weekly cycles of iv streptozotocin (STZ). There were no objective responses. Four patients progressed by the third week of therapy and an additional eight by the fourth week. Toxicity was acceptable especially in view of the extensive prior therapy. STZ is unlikely to be of benefit in combination chemotherapy regiments for SCBC.


Myocardial collagen, the effects of right ventricular hypertrophy and its involution induced by changes in atmospheric pressure. Right ventricular hypertrophy induced by hypobaric conditions was accompanied by an increased collagen mass. Left ventricular collagen was unaffected. Right ventricular mass remained unchanged after involution of right ventricular hypertrophy on return to normobaric conditions. Dietary restriction limited the size and collagen mass of both ventricles.


Ultrastructural, cyto- and biochemical observations during turnover of plasma membrane in duck salt gland. The mechanism of plasma membrane turnover was investigated using the duckling salt gland as a model system. Feeding fresh water to salt-stressed ducklings results in a decrease in the Na, K-ATPase in salt gland to non-stressed levels in about 7 days, as measured by ATP hydrolysis and 3H-ouabain binding. Electron micrographs reveal that this is accompanied by a decrease in plasma membrane infoldings on the basal and lateral borders of gland secretory cells. Simultaneously there is an increase in filamentous material and a rise in acid phosphatase and peptidase activities in these cells. Cytochemistry shows that the acid phosphatase activity is mostly associated with the basal or basolateral regions of secretory cells. These ovservations could indicate that the removal of plasma membrane components is accomplished by internalization and digestion within the secretory cells.


Eelectrocardiographic correlates of ultrasonically increased septal, left ventricular posterior wall and left ventricular internal dimensions. The electrocardiograms (ECG) of 64 subjects who exhibited an echocardiographically demonstrable increase in thickness of the interventricular septum and left ventricular posterior wall (Group 1, 22 patients), isolated left ventricular internal dimension (Group 2,26 patients), combined wall thickness and chamber diameter (Group 3, 2 patients), and septal thickness, (Group 4, asymmetric septal hypertrophy, 14 patients) were reviewed in order to determine sensitivity of ECG criteria for the diagnosis of left ventricular hypertrophy (LVH) proposed in 1949 by Sokolow and Lyon (13), in 1968 by Romhilt and Estes (14), and in 1973 the New York Heart Association (15). Relative sensitivity of the three methods was as follows: Total group, NYHA (77%) greater than Sokolow and Lyon (67%) greater than Romhilt and Estes (58%); Group 1, NYHA (91%) greater than Sokolow and Lyon (73%) greater than Romhilt and Estes (54%); Group 2, NYHA and Sokolow and Lyon (65%) greater than Romhilt and Estes (61%); Group 4, NYHA (79%) greater than Sokolow and Lyon (64%) greater than Romhilt and Estes (57%). We conclude that 1)ECG criteria of the NYHA for the diagnosis of LVH correlate best with an increase of ultrasonically determined septal, left ventricular posterior wall or left ventricular internal dimensions when compared with voltage criteria of Sokolow and Lyon and the point score system of Romhilt and Estes; and 2) isolated increase of left ventricular internal dimension, in the absence of thickened septum or posterior left ventricular wall, frequently results in ECG criteria compatible with the diagnosis of LVH.


[Rate of structural renaturation of curare-like polypeptide from cobra venom]. After breakage of their four disulfide bonds, snake curare-like toxins recover spontaneously their native conformation at different rates depending on the number of residues in the sequence.


[Histochemical characterization of the tonic fibers in the flexor carpi radialis muscle of frogs (Rana temporaria, L.)]. In the flexor carpi radialis muscle of the Frog, extra-fusal muscular fibres with histochemical characteristics of tonic fibres were identified by reactions for enzyme activities, especially for myofibrillar adenosine triphosphatase. This evidence suggests no difference between the sexes in the organization of this muscle, in spite of a strongly marked sexual dimorphism.


Influence of endotoxin on myocardial calcium transport and the effect of augmented venous return. The influence of endotoxin shock and of experimentally increased venous return during endotoxin shock on myocardial vesicular calcium uptake and calcium stimulated ATPase activity was investigated. Vesicular calcium uptake was depressed (p less than 0.01) from 0.9 mumoles/mg protein/min to 0.3 mumoles/mg/min after 5 hr of endotoxin shock. Control ATPase did not differ between endocardial surface and epicardial surface. This was accompanied by a depressed (p less than 0.01) ATPase activity from 1.2 mumoles Pi/mg/min at the endocardial surface, and to 0.9 mumoles Pi/mg/min at the epicardial surface. A femoral arteriovenous shunt was used to increase venous return by 313 +/- 71 ml/min (approximately 17 ml/kg) during the shock period. Vesicles from AV shunted animals after endotoxin were capable of normal calcium uptake and normal ATPase activity. Results suggest that myocardial depression during endotoxin shock is more severe on the endocardial surface and is caused by depressed vesicular calcium uptake secondary to depressed ATPase activity. Furthermore, this depression may be avoided by maintenance of an adequate venous return.


Diastolic left ventricular pressure-volume and stress-strain relations in patients with valvular aortic stenosis and left ventricular hypertrophy. Left ventricular (LV) chamber and myocardial stiffness were determined in 17 patients, four subjects with normal LV function and 13 subjects with valvular aortic stenosis and concentric myocardial hypertrophy, using simultaneous catheter micromanometry and LV cineangiography. Pressure (P), volume (V), and wall thickness (h) were measured. Variability in both chamber and myocardial stiffness parameters was found with five of the aortic stenosis patients (Group 1, left ventricular end-diastolic pressure = 15 +/- 2 (SEM) mm Hg) exhibiting normal values for end-diastolic dP/dV and dP/dV/V, for chamber stiffness constants (a,a') derived from P-V and normalized P-V relations, respectively, for end-diastolic myocardial elastic stiffness (ES or EE, where S = spherical model and E = ellipsoidal model) at the midwall of the minor axis circumference, and for the myocardial stiffness constants (KS or KE) of the circumferential stress-strain relation. Eight other patients with aortic stenosis (Group II, left ventricular end-diastolic pressure = 20 +/- 3 (SEM) mm Hg) exhibited significant increases in end-diastolic dP/dV,dP/dV/V,ES and EE and a tendency for increase in the chamber stiffness constants (a,a') and myocardial stiffness constants (KS, KE). These observations suggest that concentric increase in muscle mass (increase in wall thickness/minor axis radius ratio and wall volume/chamber volume ratio) is an important determinant of elevated mid- and late diastolic pressures in patients with valvular aortic stenosis, while concurrently mitigating increases in both systolic and diastolic wall stress. In some patients with aortic stenosis, however, diastolic filling pressures are elevated more severely, not only as a result of concentric hypertrophy, but also in response to augmented muscle stiffness. Reversibility of increased ventricular diastolic stiffness and elevated filling pressures was documented as concentric hypertrophy regressed post-aortic valve replacement in one patient, suggesting that fibrosis is not invariably the cause of enhanced myocardial stiffness in this secondary and compensatory form of hypertrophy.


The Croydon workshop for parents of pre-school mentally handicapped children. A multidisciplinary team of professionals met the parents of 25 pre-school handicapped children for a series of evening meetings to discuss how professionals and parents can help the development of the child. The workshop was exceptionally well-attended by parents who received practical advice from both the professionals and other parents. Multidisciplinary parents workshops appear to be an effective community service for the parents of handicapped children.


Family life with a handicapped child: the parents speak. This paper summarizes the main points made in 60 interviews with parents of handicapped children living in one City. Its aim is to provide information from the consumers of the professional services which may have relevance for future planning.


Phospholipid composition of Down's syndrome and normal platelets. 1. Total and individual phospholipids were quantitated in platelets isolated from normal and Down's syndrome (D.S.) subjects. Phospholipids were extracted from isolated platelets, separated by thin layer chromatography and quantitated by measurement of fluorescence of the compounds using a thin layer chromatogram scanner. The total amount of phospholipid was similar in D.S. and control subjects. The amount and percent composition of phosphatidyl serine, phosphatidyl ethanolamine, phosphatidgl inositol, phosphatidyl choline and sphingomyelin was also similar in D.S. and normal platelets. 2. The studies were undertaken to determine if the decrease in Na+/K+ AtPase activity observed in D.S. platelets was associated with alteration in phospholipid composition. The present report would indicate that no major differences in phospholipid composition are present in D.S. platelets that would account for the observed decrease in Na+/K+ AtPase activity.


Effects of macromolecular compounds on human leukocyte beta-galactosidase activity. The enzyme properties of leukocyte beta-galactosidase and the effects of macromolecular compounds on its activity were studied in detail. It was demonstrated that various macromolecular compounds markedly influence the activity of beta-galactosidase. These phenomena should be taken into account when considering the biochemical abnormalities of tissues of patients with mucopolysaccharidoses and mucolipidoses.


The antibody response of institutionalized Down's syndrome patients to seven microbial antigens. An earlier cross-sectional study had revealed that institutionalized Down's syndrome (DS) patients possessed much lower titres of hepatitis B surface antibodies (anti-HBs) than did their non-Down's (ND) counterparts. In an attempt to determine whether DS patients were generally deficient in humoral antibody response, the inmates of an institution for the mentally retarded (110 DS, seventy-eight ND) were immunized with tetanus, diphtheria (toxoids), influenza A, influenza B (inactivated vaccines), measles, mumps and rubella (attenuated vaccines), and tested for their antibody responses. The DS and ND groups did not respond differently to any of the seven antigens. Furthermore, there was no general relationship between the anti-HBs titres of inmates and their capacity to respond to the defined antigenic stimulus of any of the seven antigens. From these results it is apparent that a general humoral deficit in the DS group cannot explain their tendency to possess much lower anti-HBs titres than their ND counterparts upon becoming infected with the hepatitis B virus. When the antibody status and responses to immunization of the inmates who possessed anti-HBs were compared with those who had chronic HBsAg antigenaemia, there was no significant difference between the groups.


Phase I studies on chlorozotocin.

Phase I studies on chlorozotocin. A phase I investigation of chlorozotocin, a new-water soluble chloroethylnitrosourea, was undertaken to define its pharmacologic effects in man. Forty-three patients received single intravenous doses ranging from 5 to 175 mg/m2 every 6 wk. No signs of toxicity were observed at doses of under 120 mg/m2, but thrombocytopenia occurred at higher doses. The thrombocytopenic nadir appeared to be dose-dependent and occurred 4 wk after treatment. Platelet transfusions were required in 2 patients who had previously received intensive chemotherapy. No significant leukopenia occurred. A mild reversible and delayed elevation of hepatic transaminases was found in 25% of courses of 120 mg/m2 or more. No renal toxicity was observed and gastrointestinal toxicity was mild. Investigation of clinical pharmacology revealed a rapid triphasic plasma clearance with initial t1/2S of 3, 15, and 30 min. The concentration of N-nitroso intact drug at 1 hr was 10% of the initial peak level. Renal excretion accounted for half of the dose. No significant concentration of N-nitroso intact or radiolabeled drug was detected in the cerebrospinal fluid of 2 patients in whom it was examined. There were objective signs of therapeutic activity in 5 patients, 3 of whom had melanoma. Based on these studies, the recommended dose for phase II investigation of chlorozotocin is 120 mg/m2 every 6 wk.


Acneform lesions in Becker's nevus.

Acneform lesions in Becker's nevus. Three cases of Becker's nevus are described in which acneform lesions were confined strictly within the limits of the hyperpigmentation. The combined phenomenon has not been reported previously, and we conceive it to be either a rare variant of the anomaly or a still rarer coincidence of Becker's nevus and nevus comedonicus.


Adverse effects from topical steroids.

Adverse effects from topical steroids. The introduction of topical corticosteroids greatly expanded the practitioner's dermatologic armamentarium and at the same time augmented the list of local and systemic iatrogenic diseases with which he must deal. A recent review of the literature was undertaken to determine the adverse effects seen with topically administered corticosteroids. These were found to be arbitrarily assignable to eight categories: pilosebaceous, atrophic, therapeutic, immunologic, cosmetic, endocrinologic, oculonasal, and allergic. A brief discussion of the individual entities seen in each category as well as the etiology, when known, and treatment, if any, are included in this discussion.


Structural chromosome abnormalities in Down syndrome: a study of two families. Two families were ascertained through a proband with Down syndrome and a structural rearrangement involving two chromosomes 21. It is suggested that in one patient the chromosome is an isochromosome formed by misdivision of the centromere of a maternal telocentric chromosome 21 and that in the other a Robertsonian translocation involving chromosome 21 was inherited from the mother, who is a 46,XX/46,XX, -21,+t(21q21q) mosaic. The origin of the mosaicism is discussed and considered to be likely to be the result of breakage and reunion at the chromatid, rather than the chromosome, level.


The origin of the extra chromosome 21 in Down syndrome. Studies of fluorescent variants and satelite association in 26 informative families. Studies of fluorescence and other chromosomal variants were informative in 26 out of 72 families. Maternal  nondisjunction was found in 19 and paternal in 7 cases. Satellite association studies of these parents and 94 controls from the same age group showed a highly significant increase in the satellite association index (AI) for chromosome 21 in the parents where the nondisjunctional event had taken place. The AI was also higher for chromosome 14. In addition, the parents who produced the normal gametes had significantly higher AI's for some acrocentrics than the controls. Exogenous factors increasing satellite association cannot be ruled out. The number of 21-21 association was significantly increased in the parents with nondisjunction in meiosis 1. The results indicate that satellite association may play a role in the etiology of Down syndrome.


Radiological findings in atrial septal defect in the middle-aged. This study comprises 83 patients who underwent surgery for atrial septal defect of secundum type (ASD II) after 40 yr of age. A follow-up study was performed on average 74 mth after surgery. Before surgery enlarged heart volume was found in 72% of the patients. Increased pulmonary vascularity was the most characteristic finding (96%), followed by enlarged central hilar vessels. The heart volumes showed best correlation with the right ventricular systolic pressure, whereas correlation with the shunt ratio was much poorer. After surgery the mean heart volume was reduced by 22% in patients who were without a residual shunt. In only one of these the volume was increased. The pulmonary vascularity remained increased in 30% after surgery. In patients with a residual shunt, the frequency of still enlarged or increased heart volume, and increased pulmonary vascularity, was considerably large than in patients with completely closed ASD.


The effects of isoproterenol on taurine concentration in the rat heart. Administration of the beta-agonist isoproterenol to rats caused a sharp increase in the influx rate of taurine into the heart, but had no effect on the rate of biosynthesis of taurine in the heart. Influx rates of other amino acids were also stimulated, the distribution ratios, (dpm/g heart)/(dpm/ml plasma), of beta-amino acids being 3--4 times higher than alpha-amino acids. The doses of isoproterenol employed induced cardiac hypertrophy, but the stimulation of taurine influx appeared to be an independent phenomenon, in that near maximum stimulation of influx was achieved prior to stimulation of ornithine decarboxylase activity. Reserpine treatment also produced stimulation of taurine influx with no effect on biosynthesis. These results indicate a close connection between increased sympathomimetic activity and increased influx of amino acids.


Studies of the hemopoietic microenvironments: effects of acid mucopolysaccharides and dextran sulphate on erythroid and granuloid differentiation in vitro. The effects of acid mucopolysaccharides (AMPS) on in vitro erythrocytic and granulocytic colony formation of murine bone marrow cells have been studied. High concentrations of chondroitin sulphate A, B and C and heparitin sulphate partly or completely inhibited the response of CFU-E to erythropoietin stimulation whereas addition of heparin, hyalyronic acid and keratan sulphate II in concentrations up to 100 microgram/ml did not elicit an inhibition of erythrocytic colony formation. The granulocytic colony formation was not significantly affected by AMPS-addition under these circumstances. Low concentrations of chondroitin sulphate A and B evoked a stimulatory effect on the CFU-E number. The synthetic polyanion dextran sulphate did not affect the erythrocytic and granulocytic colony formation. It is concluded that AMPS can affect the in vitro erythrocytic proliferation and differentiation in concentrations which do not affect the granulocytic maturation. Since stromal cells, i.e. macrophages and reticular cells, in bone marrow in vivo have the ability to produce and remove AMPS in the extravascular matrix we postulate that stomal cells may be involved in the regulation of erythroid progenitor cell maturation.


Colony stimulating factor (CSF) in human mixed lymphocyte cultures: effect on human and mouse progenitor cells. Supernatants from human two-way MLC, poor and rich in monocytes, were tested for their ability to induce colony growth of human and mouse progenitor cells in semi solid agar. Colony stimulating factor (CSF), with activity in both systems, indicated that allogenic lymphocytes require monocytes to produce CSF. Whereas human marrow showed an early kinetics of production, the liberation of CSF active on mouse marrow cells exhibited a delayed kinetics. These results, combined with other evidence, suggest that human lymphocytes produce two different factors.


Some observations on the efferent innervation of rat soleus muscle spindles. In the small segmental tail muscles of the rat beta fibres provide exclusively the dynamic fusimotor control, while gamma fibres provide exclusively the static fusimor control. The present experiments were made to investigate the fusimotor innervation of spindles in a large muscle of the rat, the soleus, and thus to determine the occurrence and significance of beta innervation in this muscle. Our results have revealed no case of beta innervation in the rat soleus. As a consequence of our experimental method, however, we would not claim that beta innervation does not exist in the soleus, only that it must play an insignificant role relative to that seen in the tail segmental muscles. Investigations of the fusimotor innervation of eight spindles were sufficiently complete to warrant detailed illustration. The number of gamma fibres ranged from two to four. In every case the slowest conducting gamma fibre was dynamic. However, the conduction velocity spectra for the static and dynamic gamma fibres to rat soleus overlap to such an extent that it is impossible to use conduction velocity as the sole guide to functional gamma fibre classification. The pooled results from the eight spindles fully investigated provide a ratio of static to dynamic gamma fibres of approximately 1:1. Other evidence discussed in the paper suggests that in the muscle nerve the ratio is considerably higher. These differences are reconciled if the dynamic gamma fibres branch more profusely and innervate more spindles than do the static gamma fibres.


Increased cell membrane permeability to Na+ and K+ induced by thyroid hormone in rat skeletal muscle. Thyroid hormone (T3) increased Na+ dependent respiration accompanied by an increase in NaK-ATPase activity. Administration of T3 increased intracellular K+ concentration and Na/K ratio in thyroidectomized rats, and the Na+ efflux rate constant incubated in oxygenized Na+, K+-Ringers in euthyroid rats. However, the magnitude of the changes in intracellular K+ concentration was modest or invisible in comparison to the changes in QO2(t) and NaK-ATPase activity. The Na+ and K+ efflux rate constants in K+-free +ouabain Ringers were increased by T3 in both thyroidectomized and euthyroid rats. Thus, thyroid hormone stimulates not only Na pump but also the permeability of cell membrane to Na+ and K+. The both effects might contribute to the thyroid thermogenesis.


Salpingostomy by microsurgery.

Salpingostomy by microsurgery. Salpingostomy for hydrosalpinx was carried out using a microsurgical technique. The postoperative patency rate was over 90%. Among the 41 patients followed for more than 1 year, 29% have had one or more intrauterine pregnancies and 27% have had live births. The ectopic gestation rate was 12%. All of the tubal gestations occurred in the 1st postoperative year whereas 60% of the intrauterine pregnancies occurred after the 1st year, suggesting a degree of restoration in the mucosa and the musculature of the oviduct with the re-establishment of patency and the passage of time.


Effect of prolactin on the glycolytic metabolism of spermatozoa from infertile subjects. The effect of prolactin on the glycolytic metabolism of spermatozoa from married infertile men whose semen had spermatozoal concentrations of less than 40 million/ml was studied. In a respirometry system, spermatozoa were incubated with L-U-14C-glucose as substrate. Glucose utilization and production of 14C-lactate by the spermatozoa in the presence of 100 ng of ovine prolactin were greater than in the absence of the hormone. The production of pyruvate was 2.5 times less than the production of lactate wehn spermatozoa were incubated in the absence of prolactin and 1.6 times less in the presence of prolactin. These results indicate that, in spite of having an increased glycolytic metabolism, spermatozoa from oligospermic men are are able to show additional glucose utilization in the presence of prolactin.


Renal hypertrophy in experimental diabetes: a comparison to compensatory hypertrophy. The kidney growth seen after the induction of experimental diabetes in rats has been compared to the compensatory renal growth after one-sided nephrectomy. After five days the kidney weight had increased from 650 +/- 15 mg in a group of controls to 778 +/- 21 mg in diabetic rats, and 764 +/- 17 mg in unilaterally nephrectomised rats. The increased weight was in both groups reflected in an increased DNA content and increased RNA/DNA and protein/DNA ratios. In a group of rats made diabetic and nephrectomised at the same time, kidney weight increased to 953 +/- 22 mg after five days. When rats were unilaterally nephrectomised after 20 days of untreated diabetes, compensatory growth was much more pronounced than in non-diabetic rats, kidney weight rising from 780 +/- 21 to 1144 +/- 39 mg in five days. Similarly, in rats with established compensatory renal hypertrophy, diabetes induced a very rapid growth of the remaining kidney (1226 +/- 46 mg after five days).


Immune responses of diabetic animals. Comparison of genetically obese and streptozotocin-diabetic mice. The blastogenic responses of lymphocytes from chemically-induced (streptozotocin) and genetically-diabetic C57B1/6J (ob/ob) obese mice were assessed using mixed-lymphocyte cultures (MLC) and mitogens selective for thymus-derived (T cell) and bone marrow-derived (B cell) lymphocytes. Splenic lymphocytes from obese and normal C57B1/6 mice exhibited similar responses to the nonspecific T and B cell mitogens, Concanavalin A (Con A) and E. coli lipopolysaccharide (LPS), respectively. A small (25%) depression of the blastogenic response in MLC was observed for lymphocytes from obese mice. The generation of cytotoxic T cells in vitro in response to trinitrobenzene sulphonic acid (TNP)-modified syngeneic spleen cells was the same for normal and obese mice. In contrast, splenic lymphocytes from 7-14 day streptozotocin-diabetic mice had lower (56-60%) proliferative responses in MLC. The generation of cytotoxic effector cells in vitro was lower for spleen cells for spleen cells from 22-day streptozotocin mice, although blastogenic responses in MLC were not depressed. The insulin-deficient streptozotocin mice appear to have a depression of some thymus-derived cell functions that may be associated with streptozotocin rather than the diabetic state. Direct immunosuppressive effects of streptozotocin are indicated by the marked decrease in the number of lymphocytes in the thymus, lymph nodes, and spleen.


[Comparison between vectocardiogram and echocardiogram in the diagnosis of left atrial enlargement (author's transl)]. The purpose of this study was to evaluate the influence of atrial enlargement on P wave abnormalities in "primary" (mitral stenosis) and "secondary" (left ventricular involvement) forms of left atrial overload. Echocardiograms and Frank Vectorcardiograms were obtained from 42 subjects, including 12 patients with mitral stenosis (group I A), 4 with mitral insufficiency (group I B), and 26 with left ventricular disease. Good correlations were founded between left atrial dimension and the following vectorcardiographic criteria: magnitude of the positive P vector in lead Z, sum of the positive P wave in leads X and Z, P positive duration/PR segment ratio in lead Z. No specific difference has been found in the vectorcardiographic abnormalities of the "primary" and "secondary" left atrial enlargement. Separate analysis revealed that P wave duration or amplitude changes can be proposed in group I A as specific and sensible criteria of left atrial enlargement. In contrast, the same criteria are highly unspecific when applied to the patients with left ventricular disease, because they can reflect the influence of other variables (left atrial pressure, intra-atrial conduction defects). Comparison of our results with those of other studies of P wave analysis did not demonstrate the superiority of the vectorcardiogram over the conventional electrocardiogram in the diagnosis of left atrial enlargement.


Effects of chlorpromazine hydrochloride and its metabolites on Mg2+- and Na+,K+-ATPase activities of canalicular-enriched rat liver plasma membranes. Recent experiments in primates indicate that the phenothiazine drug chlorpromazine hydrochloride inhibits both bile salt-dependent and -independent bile flow in a predictable fashion. Because a significant fraction of bile salt-independent bile flow is postulated to depend upon the activity of a canalicular membrane Na+,K+-ATPase, we have examined the effects chlorpromazine hydrochloride and its metabolites on the ATPase activities of canalicular-enriched rat liver plasma membranes. Chlorpromazine inhibited the activities of both Mg2+- and Na+,K+-ATPases with a linear dose-response relationship between 10 and 100 micronM. The inhibition of Na+,K+-ATPase was pH dependent, showing a maximal inhibition at pH 7.8. Over the pH range 7.0 to 8.2, the inhibition was significantly reduced with the addition of glutathione and was augmented under experimental conditions (ultraviolet irradiation and peroxidase-H2O2) that promote the formation of the chlorpromazine semiquinone free radical. The 7-hydroxychlorpromazine metabolite was as active an inhibitor as the parent drug; however, two sulfoxide metabolites, chlorpromazine sulfoxide and 7-hydroxychlorpromazine sulfoxide, were less effective inhibitors of Na+, K+-ATPase. Our data are consistent with the hypothesis that chlorpromazine cholestasis may be a result of a direct toxic effect on the ATPase activities of hepatic canalicular membranes. Our results further suggest that if chlorpromazine cholestasis occurs through an interaction with canalicular membrane ATPases, the degree of cholestasis may well be influenced by the extent of the conversion of the drug to its more active (free radical) or minimally active (sulfoxide) metabolites and by the local environment (pH and glutathione concentration) of the canalicular membrane.


[Experience with the tupla-clip for tubal sterilization by laparoscopy (author's transl)]. For the last 18 months a plastic clip made of polyoxymethylene was used for tubal sterilization by laparoscopy. The clip was used in 140 cases. This report reviews 106 cases. One pregnancy occurred early in this series. There were no other complications. The advantages of the method are the short operating time and the elimination of intra-abdominal damage by heat. Reversibility of the method has not yet been assessed.


[Report of a case of primary abdominal pregnancy (author's transl)]. A case of primary abdominal pregnancy is described. The diagnosis was suspected by laparoscopy and confirmed by the laparotomy in the same operative session. The microscopic examination corroborated the diagnosis. The patient was discharged in good condition on the 12. post-operative day.


Nuclear mutation increases streptomycin and spectinomycin sensitivity in Chlamydomonas. A spontaneously arising nuclear mutation, ss-1, has been identified in Chlamydomonas reinhardtii that decreases both streptomycin and spectinomycin resistance levels about 10-fold after its introduction into all wild-type, streptomycin-resistant and spectinomycin-resistant strains examined. The mutations for resistance map to nuclear and uniparentally inherited (chloroplast) loci. In contrast, no modification of erythromycin resistance was detected after introducing ss-1 into wild-type strains or into strains carrying nuclear or uniparentally inherited erythromycin-resistance mutations. We suggest that ss-1 affects the small subunit of the chloroplast ribosome because others have shown that streptomycin and spectinomycin resistance in C. reinhardtii are associated with this subunit, whereas erythromycin resistance is associated with the large subunit. ss-1 shows no linkage with the nuclear locus for streptomycin resistance.


Reconditioning aging muscles.

Reconditioning aging muscles. Weakness or stiffness of key posture muscles can cause much of the disability seen in elderly patients. Too much tension and too little exercise greatly increase the natural loss of muscular fitness with age. A systematic program of exercise, stressing relaxation and stretching of tight muscles and strenghthening of weak muscles, can improve physical fitness. The program must be tailored to the patient, starting with relaxation and gentle limbering exercises and proceeding ultimately to vigorous muscle-stretching exercises. Muscle aches and pain from tension and muscle imbalance are to be expected. Relaxation relieves tension pain, and strengthening weak muscles and stretching tight muscles will correct muscle imbalance. To prevent acute muscle spasm, the patient should avoid excessive exertion and increase exercise intensity gradually.


[The value of radiographic examinations of the thorax in acute myocardial infarction (author's transl)]. The degree of pulmonary congestion seen on chest radiographs taken in the intensive care unit was compared with the end-diastolic pulmonary artery pressure in 65 patients with recent transmural myocardial infarcts without cardiogenic shock. There was good correlation between the radiographic appearances and the end-diatrolic pulmonary artery pressure, so that the severity of left heart insufficiency could be diagnosed radiologically at an early stage. Errors were due to the time interval which must pass before the radiographic appearances approximate the haemodynamic changes. This resulted in a diagnostic "phase lag" (following a rise in end-diastolic pulmonary artery pressure) or in a post-therapeutic "phase lag" (after a fall in end-diastolic pulmonary artery pressure). Repeated examinations allow one to recognize changes in the haemodynamics. One should pay particular attention to the features of pulmonary venous and pulmonary arterial hy


[Coronary flow measurements with cine densitometry. II. Quantitative determinations of flow velocities and flow volumes through severe coronary stenoses during rest (author's transl)]. The normal flow values through healthy or moderately narrowed coronary arteries, which had been determined by a new method of cine radiography, together with a computer, were published in this Journal in 1976. In the present paper we report the results of further studies of flow velocity and flow volume through high-grade coronary artery stenosis of severity III and III--IV. Compared with the results obtained on normal and mildly stenosed arteries, measurements at rest through severely stenosed coronaries showed an almost constant flow. The first results on aorto-coronary vein graft measurement are described.


[The diagnostic value of kymography in aortic isthmus stenosis (author's transl)]. The value of kymography was studied in 133 patients in whom a coarctation had been demonstrated angiographically, operatively or pathologically; its value was compared with conventional radiographic examinations. The difference in pulsation between the pib notching, dilatation of the ascending aorta and increased size of the left ventricle on plain films make it possible to diagnose an haemodynamically significant coarctation without any invasive methods. In 124 haemodynamically significant coarctations (93%), the kymogram showed increased pulsation of the dilated left subclavian artery. In nine haemodynamically insignificant or mild coarctations (7%), this sign was absent. Since the radiographic signs are to some extent reversible, it is possible to evaluate the results of surgery by plain chest films and the kymogram.


[The cest radiograph in Wegener's granuloma (author's transl)]. Wegener's granuloma belongs to the so-called collagenoses. It is characterized by focal vasculitis, necrotising granulomatous processes in the upper air passages and lung, as well as focal glomerulitis with swelling of the capsule and the formation of granulomatous glomerular lesions. In order of frequency the lungs, paranasal sinuses, pharynx, kidneys, joints, skin, eyes, ears, the heart and nerve tissue may be involved. Recently, Wolf and Fischbach (1976) reported six cases of Wegener's granuloma in this Journal, with radiological and pathological changes in the lungs. Typical changes which may be found radiographically are illustrated in the present paper based on four histologically confirmed cases of this relatively rare condition.


[Unilateral hilar enlargement in sarcoidosis--case report and review of the literature (author's transl)]. Case report on unilateral hilar enlargement in sarcoidosis is given and the literature is reviewed. The diagnostic kriteria of angiography in these cases is discussed. Key words: Sarcoidosis, unilateral hilar enlargement, bronchialarteriography, lung szintigramm.


[Bronchial arteries in interstitial pulmonary fibrosis (author's transl)]. 6 autopsy lungs with interstitial pulmonary fibrosis were subjected to comparative angiographical and morphological investigations of bronchial arteries. During the final stage of interstitial pulmonary fibrosis, bronchial arteries were increased in number, and so were their anastomoses with pulmonary arteries. Extensive formation of broncho-pulmoary anastomosis as well as proliferation and formation of new bronchial arteries, is associated with interstitial chronic inflammation and fibrosis in the lung. The findings explain functional interconnections between the vessels of the systemic and the pulmonary circulations by direct artery-to-artery anastomoses, with a negative influence on pulmonary hypertension.


[Angiographic findings in a new method of vascular anastomosis (author's transl)]. The first angiographic findings with a new method of end-to-end anastomosis of small arteries are reported. The other-wise difficult anastomosis of small vessels is accomplished by the use of scew-ended flanges. So far the method has been used successfully on 15 dogs; there were no post-operative stenoses or other complications.


[Therapeutic embolisation of visceral arteries. Animal experiments, early clinical experience and introduction of a new embolising material (author's transl)]. In order to compare the effectiveness and suitability of the most commonly used embolising materials--autologous blood clot, either pure or modified by epsilon-amino-caproic acid, gelatin, or fibrin foam--a series of experiments was carried out using the renal arteries of rabbits. Our own experience and the information in the literature indicate that there is at present no optimal and lasting material for embolisation. A new radioopaque for am consisting of a silicone elastomer has been found to be superior to all other materials, without any evidence of complication arising from it. Contrary to present views, it has been shown that the effect of embolisation is best demonstrated scintigraphically; angiography immediately after embolisation does not provide the necessary informatin (spasm, further thrombosis). The results of animal experiments and toxicity studies indicated that the substance can be used as suitable embolising material in patients, provided the indications are strictly controlled.


[Results of percutaneous transhepatic portography (author's transl)]. The technique, indications, contraindications and complications of percutaneous transhepatic portography based on the experience of 300 cases are described. The advantage of this method is discussed and compared with transjugular and transumbilical portography.


[Changes in liver haemodynamics after mesenterico-caval dacron prosthesis anastomosis ("H-shunt") in portal hypertension (author's transl)]. In fourteen patients with portal hypertension and bleeding from oesophageal varices, mesentericocaval anastomoses using a dacron-velour prosthesis ("H-shunt") were carried out. Evaluation of the haemodynamics showed: 1. Comparison of 13 pre- and post-operative angiograms showed an haemodynamicically effective shunt in 12 and a stenosed shunt in one. 2. The aim of relieving the portal circulation while maintaining antegrade portal flow was achieved in six patients. 3. In six patients, the portal vein was not seen in a splenic or mesenteric portogram despite an open shunt. Functionally, this corresponds to a porto-caval anastomosis. 4. Pressure reduction in the portal circulation can be so marked as to obtain retrograde portal flow from the hepatic artery. 5. If the portal blood flow through the liver disappears, the hepatic artery may obtain additional flow from the superior mesenteric artery (reversed flow in the gastroduodenal artery). 6. Increased arterial flow to the liver is made possible by a reduction in intrahepatic resistance. The role of collaterals, either arterio-portal shunts or shunts between the sinusoids, is discussed. 7. The hepato-fugal collateral circulation was reversed in twelve patients.


Percutaneous transhepatic drainage catheter: a valuable therapeutic aid in obstructive jaundice. Percutaneous decompression of the biliary system is a useful and well established procedure which can be carried out with negligible risk and may be of great benefit to the patient. The main indications are pre-operative decompression and primarily decompression of patients with inoperable obstructive neoplastic disease. In order to provide drainage, the catheter should be advanced under fluoroscopic control into one of the major radicles ore preferably into the distal common duct beyond the point of obstruction. This ideal internal drainage into the duodenum was accomplished in the reported patient with sclerosing cholangitis who was markedly improved by this procedure.


[The radiological diagnosis of hydatid cysts of the liver (author's transl)]. The radiological features of cystic and alveolar echinococcal disease of the liver is described, based on 17 patients. Conventional radiographs, liver ultrasound and liver angiography are described.


[Hepatobiliary sequential-scintigraphy--progress in the evaluation of postcholecystectomy complains (author's transl)]. The hepatobiliary sequential-scintigraphy is a nuclear diagnostic procedure useful in assessing the function of liver and biliary system. 40 patients with and 20 patients without complains following cholecystectomy were examined. The value of this method in establishing biliary causes for such complains is demonstrated by typical cases. Simplicity and safety in application as well as diagnostic sensitivity should place the method in the first place evaluating postcholecystectomy complains. Succeeding indirect and direct radiologic techniques then may be used systematically with benefit for the patient and physician. Results indicate, that the procedure will be essential as well in evaluation as in analysis of nature and frequency of such complains by its superior presentation of intrahepatic bilestasis and functional mechanisms of the biliary system including the sphincter of Oddi.


[Erroneous diagnoses in the retrograde demonstration of the biliary and pancreatic ducts. Results of a prospective study involving 197 examinations (author's transl)]. The results of endoscopic retrograde cholangio-pancreatography were evaluated in a prospective study of 126 patients in respect of diagnostic accuracy, as compared with surgery and clinical follow-up. The diagnosis was incorrect in 11 patients; in seven it failed to differentiate between acute and chronic pancreatitis. The discrepancies in the other four patients are described in detail, particularly with reference to prepapillary filling defects. The indications, advantages and limitations of ERCP are discussed.


Atypical pancreatic pseudocysts.

Atypical pancreatic pseudocysts. The authors present three cases of atypical pancreatic pseudocysts, occurring in the course of chronic pancreatitis: intrasplenic pseudocyst, pseudocyst in the hilus of spleen and bleeding into the pseudocyst of the tail of pancreas. Such cysts are extremely difficult, or even impossible to be recognized by conventional clinical and radiological methods. Located in the parenchyma or in the hilus of spleen, they are formed by means of fistulization along the lienorenal ligament. In every case when the chronic pancreatitis coexists with the splenic cyst, the intrasplenic localization of the pancreatic speudocyst should be taken into consideration. The cyst in the hilus of spleen may imitate intraperitoneal mass and its relation with the pancreas may be recorded angiographically only. Angiography is also the basic diagnostic method in the haemorrhage into the pseudocyst. Bleeding into the pseudocyst has a form of vascular blush appearing during arteriography. The blush increases its intensity and persists after an examination. Early diagnosis in atypical pseudocyst is vital for the patient's fate.


[The use of computer tomography in individual treatment planning (author's transl)]. Computer tomography provides the possibility of individual treatment planning. For this purpose it is necessary to obtain the matrix of linear absorption coefficient for the type of radiation used during radiotherapy. In this paper it is shown that it is necessary to obtain a cross section of the patient using rays of two different qualities. From these results and previous measurements on phantoms it is possible to calculate the density and effective atomic number. It is also shown that the most significant information is obtained from the density matrix, if ultra hard X-rays are used for radiation therapy; With this density matrix it is possible in a simple manner to take account of inhomogeneities within individual patients.


Sequential trials with new rare-earth intensifying screens. Sequential analysis of idential radiographs with calcium tungstate (CaWO4) and rare-earth intensifying screens was performed in four test situations. Most preferences in the direction of rare-earth screens were due to motion unsharpness with CaWO4 screens. Better contrast was found with rare-earth screens in one test situation in spite of low kVp exposures. Assessment of the quality of the radiographs in practical situations gave results which differed to a certain extent from what might be expected from the technical assessment of the properties of the screens.


[New aspects concerning exposure factors during direct geometrical X-ray magnification (author's transl)]. The paper is concerned with the use of rare-earth intensifying screens in order to overcome some of the exposure problems during magnification techniques using very small focal spots. Theoretical experiments and tests with biological objects indicate that the use of high intensifying screens under these circumstances is acceptable and useful. A comparison has been carried out between the results of conventional techniques without magnification and magnification techniques using very small focal spots and rare-earth intensifying screens.


The future of non-invasive medical imaging. A variety of new non-invasive imaging techniques are now at the physicians' disposal for patients' management. However, the place of each new imaging modality within the general diagnostic armamentarium is far from established. Not only comparative assessments of these are needed but also pre-defined strategies for use are required for an intelligent approach to a difficult medical problem. The increasing cost of medicine and the decreasing resources available for development enhance the necessity for such an approach. This paper offers an initial solution to the problem.


[Spieghel's hernia and its treatment].

[Spieghel's hernia and its treatment]. It is reported about 12 patients with hernias through the spigelian fascia, among them one case with a rare bilateral hernia. The cause of these hernias are congenital or acquired gaps in the fascia transversalis medial to the linea semilunaris. Mostly they are discovered below the umbilicus in the height of the linea semicircularis, lateral to the rectussheath and medial to the spigelian line. All clinical details are shown in a table (Tab. 1). There is referred about localisation, sex, age, complications before operation (e.g. incarceration), complaints of the patients and operative findings. The results correspond to those of other authors. Seldom a spigelian hernia is noticed in children. To diagnose a spigelian hernia it is very important to think of it, for the symptoms are often not very characteristically and the clinical findings misleading. At times only operation reveals the real diagnosis. The operation is often simple and remaining complaints are very seldom; we didnt see any. Sometimes the operative finding requires an extensive laparotomia and bowel resection. If one finds the abdominal wall intact, one should open the abdomen in every case.


[Neurophysiological activation and behavior during mental achievement. Comparative studies in healthy, minimally brain damage and mongoloid children]. A comparison of normal, mildly brain-injuried and mongoloid children. For psychophysiological investigations in  young or mentally disturbed children a new methodical approach was developped. In children with minor nervous dysfunction (aged 6--8;3 years) and mongoloid children (5;6--11 years) as well as in younger and older normal children (3;9--5;5 resp. 5;6--7;6 years) the changes of EEG basic rhythm, heart rate and behaviour during defined mental work were observed. Each group consisted of 18 children. The physiological data were analyzed by computer. The main results are summarized in the following points: 1) The request of mental activity with increasing task difficulty causes increasing neurophysiological and behavioural activation up to an optimal value at the highest degree of individual efficiency. 2) Overstrain leads to a deterioration of activation and consecutively to a loss of motivation, especially in the handicapped children. 3) Alpha waves of the EEG can be unterstood as an index of selective stimulus inhibition or "concentration". In the normal children the amount of alpha waves during mental work was very significantly higher than in both the groups of the handicapped children. 4) Further differences in EEG, heart rate and behaviour between the 4 groups support the understanding of the psychic features in Down's syndrome and in the syndrome of minor nervous dysfunction at a physiological level.


[Importance of gynecological laparoscopy in diseases of the sigmoid in the aging]. During the last six years we have performed laparoscopy in 80 women older than 60 years of age. In 22 of these, the barium enema showed pathological colonic conditions (mostly diverticulosis or diverticulitis). Despite these findings laparoscopy was used as an additional and reliable diagnostic aid to exclude concomitant gynecological diseases. Of these 22 women, 16 had normal internal genital organs so that exploratory laparotomy could be avoided. In view of the high postoperative mortality in this age group this is regarded as a considerable advantage. In three women an additional gynecological disease requiring laparotomy was found. In two cases laparoscopic diagnosis was impossible because of extensive adhesions. In one patient the radiological diagnosis of diverticulitis was revised by laparoscopy which showed carcinoma of the sigmoid.


Insulin responsiveness of isolated perfused livers from rats with streptozotocin induced diabetes. The ability of insulin to inhibit efflux of potassium (K) and amino acid nitrogen (AAN) from perfused livers of normal and insulin deficient rats was studied. Two groups of rats with different degrees of insulin deficiency were produced by injecting varying amounts of streptozotocin. One group, classified as being moderately diabetic (MD), had fasting plasma glucose levels between 235--425 mg%, while the other group, whose plasma glucose levels greater than 425 mg%, were considered to have severe diabetes (SD). Two other groups of rats were food restricted in order to attain body weights comparable to the two groups of diabetic rats, and livers from these animals were used for control perfusions. The results indicated that the ability of insulin to suppress efflux of K and AAN from perfused livers of rats with MD was comparable to that seen in control perfusions. On the other hand, insulin could not suppress the efflux of either K or AAN from perfused livers of rats with SD. These results indicate that normal hepatic responsiveness to insulin can be lost secondary to the production of insulin deficiency.


Enzymatic studies of liver cells obtained by fine needle biopsy from mice with the obese-hyperglycemic syndrome (genesymbol ob). In an enzymatic study of livers from obese hyperglycemic mice (obob) and their lean litter mates, microdissected freeze-dried fine needle biopsies were used. Carbohydrate, fatty acid and ketone body metabolism was examined through assays of phosphofructokinase (PFK), 3-hydroxyacyl-CoA dehydrogenase (HOADH) and D-3-hydroxybutyrate dehydrogenase (HBDH). At 2 months of age the obob-mice showed a significant increase in PFK activity as compared to their lean litter mates. No such difference was found for the activity of HOADH and HBDH. An early increase in the initial glycolysis in the livers of obob-mice might thus be one of the factors responsible for the accelerated synthesis of lipids resulting in the fatty livers of the adult obob-mice. The present results corroborate previous studies indicating no major derangement in the fatty acid and ketone body metabolism in the livers of obob-mice. In a methodological evaluation of the technique for tissue sampling and preparation, freeze-dried fine-needle punctates showed significantly higher enzyme activities of PFK and HOADH than freeze-dried liver sections or crude homogenates. Freeze-drying of fine needle biopsies appears to be a technique with good preservation of the enzyme activity. This is of significance for future metabolic studies also in humans.


Acne treated with a tropical tetracycline preparation: results of a one-year multi-group study. One hundred five patients were treated effectively with topically applied tetracycline in an ethanol-water solution with n-decyl methyl sulfoxide, and observed for a year. Comedones and cystic lesions were least responsive to topical tetracycline. Side effects, which were minor, included yellow staining of facial skin and stinging after application. Two patients, one with juvenile diabetes and one with congenital nephritis, successfully used the preparation without adverse effects.


Comparative effectiveness of benzoyl peroxide and tretinoin in acne vulgaris. A controlled, randomized clinical study comparing different commercially available benzoly peroxide gels and tretinoin cream was undertaken by four investigators using similar protocols and case report forms. Results were based on lesion-count reduction over an 8-week period. No significant difference in reduction of total acne lesions was detected between the benzoyl peroxide gels and tretinoin cream. However, benzoyl peroxide was significantly better (P less than .02) in reduction of papules and as good as tretinoin in reduction of comedones. Results of this study suggest that excessive drying and peeling are not essential in the reduction of acne lesions.


Reproductive behaviours of Japanese parents of children with mongolism. The reproductive activity of Japanese mothers of mongoloid children decreases significantly after the birth of such a child, in comparison with mothers of normal children matched on the basis of maternal age. This decline of reproduction is observed more clearly among younger mothers whose fertility is thought still high.


[Hyperkeratotic vegetating form of mycosis fungoides. A case report]. A patient with hyperkeratotic and vegetating mycosis fungoides is reported. The immunologic investigations reveal a well preserved immunologic response. The clinical manifestations are considered as an hyperergic reaction of a sentizied organism.


Comparison of red and white muscles by cytophotometry of their muscle fibre populations. Samples from two red muscles (vastus intermedius and vastus medialis) and two white muscles (biceps femoris and gluteus medius) were taken from four pigs. Serial transverse sections were reacted for ATPase and NADH oxidative activity. Sections were mapped with a projection microscope so that the staining intensity of individual fibres for the two reactions could be measured with a simple microscope photometer. Transmission values at 600 nm were converted to units of 0-10 for the range from darkest to lightest staining fibres on each section to cancel variation in staining intensity between sections. The aim of the study was to use simple cytophotometry instead of subjective judgement in the categorization of different histochemical types of muscle fibres. Cytophotometry enabled clear resolution of the major fibre types (types I and II using the ATPase reaction), partial resolution of more variable characteristics (NADH oxidative activity in type I and II fibres) and no resolution of subtle subtypes (IA and IB with the NADH oxidative reaction). However, between the major fibre types, cytophotometry revealed variable numbers of fibres with transitional characteristics. There were more of these fibres in red muscles. With sections reacted for ATPase, transmission values for low magnification fields containing 100 to 200 fibres were correlated (r = -0.91) with the ratio of type I:II fibres.


Use of an extracardiac conduit in the repair of supravalvular pulmonic stenosis in a dog. Congenital supravalvular pulmonic stenosis was diagnosed in a dog. A pulmonary artery-to-pulmonary artery anastomosis of a Dacron arterial conduit restored continuity of blood flow around the obstruction in the main pulmonary artery. By the 6th day after surgery, right ventricular systolic pressure averaged 85 mm of Hg, a decrease of 90 mm from the preoperative value. Angiocardiography confirmed the patency of the conduit. At examination 6 months after surgery, the dog was clinically normal.


The teratogenic effects of 5-fluoro-2-desoxyuridine (F.U.D.R.) on the Wistar rat fetus with particular reference to cleft palate. The teratogenic effects of 5-fluoro-2-desoxyuridine on Wistar rat fetuses were studied macroscopically, microscopically and ultrastructurally. In no case did palatal shelf elevation occur, and a palatal shelf elevating force could not be demonstrated in freshly-removed fetuses. The shelves, like the connective tissues generally, showed clear evidence of depressed mucopolysaccharide synthesis. The shelves eventually ossified in the vertical position, and in some cases their free edges fused with the lateral wall of the oral cavity (palato-oral fusion). The results were consistent with the mucopolysaccharide-turgor hypothesis advanced by the author in a previous paper to account for normal shelf elevation. Phocomelia, brain and limb bud haemorrhages, gross subcutaneous oedema, hyperextension of the cranial base, fused vertebrae, detached epithelia, bizarre brain abnormalities (including some remarkable attempts at neural reconstitution) and growth and ossification of the mandible in the virtual absence of Meckel's cartilages were also noted in these fetuses.


Teaching foster grandparents to train severely handicapped persons. Five foster grandparents were taught training skills for use in their daily interactions with severely handicapped persons in an institution. Following baseline, specific teaching procedures consisting of teacher instructions, prompts, modelling, and praise were implemented. The grandparents' frequency of training three skill areas increased as the specific teaching was implemented in multiple-baseline format. The total amount of training continued as teacher instructions, prompts, and modelling were terminated and praise continued, although the grandparents spent their training time emphasizing only two of the three skill areas. Teacher presence was gradually reduced over an 11-week period, with no decrease in grandparents' frequency of training. Four of the foster grandchildren, all profoundly retarded and multiply handicapped, demonstrated progress throughout the study. Results were discussed in light of the available contributions of foster grandparents in institutional settings and maintenance of staff training.


Escherichia coli mutants deficient in deoxyuridine triphosphatase. Mutants deficient in deoxyuridine triphosphatase (dUTPase) were identified by enzyme assays of randomly chosen heavily mutagenized clones. Five mutants of independent origin were obtained. One mutant produced a thermolabile enzyme, and it was presumed to have a mutation in the structural gene for dUTPase, designated dut. The most deficient mutant had the following associated phenotypes: less than 1% of parental dUTPase activity, prolonged generation time, increased sensitivity to 5'-fluorodeoxyuridine, increased rate of spontaneous mutation, increased rate of recombination (hyper-Rec), an inhibition of growth in the presence of 2 mM uracil, and a decreased ability to support the growth of phage P1 (but not T4 or lambda). This mutation also appeared to be incompatible with pyrE mutations. A revertant selected by its faster growth had regained dUTPase activity and lost its hyper-Rec phenotype. Many of the properties of the dut mutants are compatible with their presumed increased incorporation of uracil into DNA and the subsequent transient breakage of the DNA by excision repair.


Isolation of Escherichia coli mutants with an adenosine triphosphatase insensitive to aurovertin. Energy-transducing adenosine triphosphatase (ATPase) from Escherichia coli is inhibited by aurovertin. Aurovertin-resistant mutants were generated by nitrosoguanidine mutagenesis of E. coli AN180, whose growth on a nonfermentable carbon source was blocked by aurovertin. The ATPase activity of cell extracts from 15 different mutants (designated MA1, MA2, MA3, etc.) was found to be at least 20 times less sensitive to aurovertin than that from the parent strain. The aurovertin-resistant mutants did not show cross-resistance towards a number of ATPase inhibitors including azide, dicyclohexylcarbodiimide, quercetin, 7-chloro-4-nitrobenzofurazan, and N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline. Aurovertin inhibited the energization brought about by addition of ATP to E. coli AN180 membrane vesicles; it was without effect on MA1 and MA2 membrane vesicles energized by ATP. The mutation in MA1, like other mutations of the ATPase complex, maps in the unc region of the bacterial chromosome.


Pathway of succinate and propionate formation in Bacteroides fragilis. Cell suspensions of Bacteroides fragilis were allowed to ferment glucose and lactate labeled with (14)C in different positions. The fermentation products, propionate and acetate, were isolated, and the distribution of radioactivity was determined. An analysis of key enzymes of possible pathways was also made. The results of the labeling experiments showed that: (i) B. fragilis ferments glucose via the Embden-Meyerhof pathway; and (ii) there was a randomization of carbons 1, 2, and 6 of glucose during conversion to propionate, which is in accordance with propionate formation via fumarate and succinate. The enzymes 6-phosphofrucktokinase (pyrophosphate-dependent), fructose-1,6-diphosphate aldolase, phosphoenolpyruvate carboxykinase, malate dehydrogenase, fumarate reductase, and methylmalonyl-coenzyme A mutase could be demonstrated in cell extracts. Their presence supported the labeling results and suggested that propionate is formed from succinate via succinyl-, methylmalonyl-, and propionyl-coenzyme A. From the results it also is clear that CO(2) is necessary for growth because it is needed for the formation of C4 acids. There was also a randomization of carbons 1, 2, and 6 of glucose during conversion to acetate, which indicated that pyruvate kinase played a minor role in pyruvate formation from phosphoenolpyruvate. Phosphoenolpyruvate carboxykinase, oxaloacetate decarboxylase, and malic enzyme (nicotinamide adenine dinucleotide phosphate-dependent) were present in cell extracts of B. fragilis, and the results of the labeling experiments agreed with pyruvate synthesis via oxaloacetate and malate if these acids are in equilibrium with fumarate. The conversion of [2-(14)C]- and [3-(14)C]lactate to acetate was not associated with a randomization of radioactivity.


Rabbit muscle phosphofructokinase. Modification of molecular and regulatory kinetic properties with the affinity label 5'-p-(fluorosulfonyl)benzoyl adenosine. The affinity label 5'-p-(fluorosulfonyl)benzoyl adenosine modifies rabbit muscle phosphofructokinase to the extent of one group/subunit. Modification appears to occur at a binding site specific for AMP, cyclic AMP, and ADP, i.e. those adenine nucleotides which are activators under conditions where regulatory kinetic behavior is obtained. The consequences of the modification are consistent with the model proposed previously for correlation between the pK of specific ionizable groups, regulatory kinetic behavior, ligand binding, and the reversible cold inactivation of the enzyme (Frieden, C., Gilbert. H. R., and Bock, P. E. (1976) J. Biol. Chem. 251, 5644-5647). Thus, the modification shifts the apparent pK of the essential ionizable groups from 6.9 to 6.4 at 25 degrees C, with the result that regulatory kinetic behavior at pH 6.9 and 25 degrees C is lost. Furthermore, the apparent affinity of a site (other than the active site) for ATP, as measured by ATP-dependent quenching of intrinsic protein fluorescence at pH 6.9 and 25 degrees C, is decreased by the modification. Regulatory kinetic behavior for both substrates is obtained with the modified enzyme at a lower pH, consistent with the downward shift in the pK of the ionizable groups, but sensitivity to cAMP activation is abolished by the modification. The loss of regulatory kinetic behavior upon modification of sulfhydryl groups does not appear to be the same as that due to modification by the affinity label.


Heart phosphofructokinase. Allosteric kinetics following affinity labeling modification of the enzyme by 8-[m-(m-fluorosulfonylbenzamido) benzylthio] adenine. An adenine analog 8-[m-(m-fluorosulfonylbenzamido)benzylthio]adenine (FSB-adenine) reacts covalently with sheep heart phosphofructokinase. Under conditions optimal for allosteric kinetics the modified enzyme is less sensitive to inhibition by ATP and insensitive to activation by AMP, cyclic AMP, and ADP. The concentration of fructose-6-P necessary for half-maximal activity is markedly decreased, while the cooperativity to the same substrate is not changed under the same conditions. The modified enzyme is more stable at pH 6.5 when compared with the native enzyme. Changes in the allosteric kinetics of the enzyme are proportional to the extent of modification reaching maximal effect when 3.2 mol of the reagent were bound/mol of tetrameric enzyme. Affinity labeling of the enzyme by the adenine derivative does not affect significantly the catalytic site. This is evidenced by the demonstration that under assay conditions optimal for Michaelian kinetics neither the Km for ATP nor for fructose-6-P is significantly changed following chemical modification. Maximal activity of the modified enzyme was 60% of the native enzyme. ADP gives the best protection, while AMP gives less protection against modification by the reagent. ATP slows the rate of the reaction and causes a slight decrease in maximum binding of the reagent to the enzyme. Modification of the enzyme caused a marked reduction of AMP and ADP binding. The evidence indicates that the modified site is a nucleotide mono- and diphosphate activation site.


Human plasma carboxypeptidase N. Isolation and characterization. Human plasma carboxypeptidase N has been purified 2,600-fold from pooled, outdated plasma in a 30% yield. Isolation was accomplished by chromatography on DEAE-cellulose and on a p-aminobenzoyl-L-arginine-Sepharose 6B affinity column. Carbohydrate accounts for 17% of the weight calculated from its amino acid and carbohydrate composition. The enzyme appears to consist of three subunits of Mr = 83,000, 55,000, and 49,000 and contains a significant amount of bound zinc. Purified enzyme preparations are very sensitive to proteolytic degradation but are stable for at least 3 months at 4 degrees.


Filamin-actin interaction. Dissociation of binding from gelation by Ca2+-activated proteolysis. Chicken gizzard filamin has been digested with purified Ca2+-activated protease. The subunits of (Mr = 250,000) of the protein are cleaved asymmetrically into two fragments, heavy merofilamin, Mr = 240,000, and light merofilamin, Mr = 9,500. Digestion is complete at substrate to enzyme ratios of 100:1 and requires Ca2+ concentrations in excess of 0.3 mM. Heavy merofilamin binds to F-actin as evidenced by cosedimentation with F-actin, by direct observation under the electron microscope, and by its ability to inhibit actin activation of heavy meromyosin ATPase. Heavy merofilamin does not form a gel when mixed with actin, except at very low concentrations of KCl. Thus, actin binding and gelation are separable activities of filamin. We speculate that Ca2+-stimulated proteolysis may play a role in the regulation of actin-filamin interactions.


Activation of the rabbit polymorphonuclear leukocyte membrane "Na+, K+"-ATPase by chemotactic factor. Addition of the synthetic chemotactic factor, formyl-methionyl-leucyl-phenylala-nine (F-Met-Leu-Phe) to medium containing magnesium, sodium, and potassium results in a doubling of the "Na+, K+"-ATPase activity of the plasma membrane fraction from polymophonuclear leukocytes (PMN). This activation is sensitive to ouabain inhibition and is dose dependent, maximal activity occuring at 10(-9)MF-Met-Leu-Phe. Equivalent activation was observed with the nonformylated derivative Met-Leu-Phe at 10(-9)M. The dipeptide, carbobenzoxy-methionylphenylalanine, which acts as an antagonist for F-Met-Leu-Phe, prevents the stimulation of the "Na+, K+"-ATPase by F-Met-Leu-Phe.


Further characterization of HeLa S3 plasma membrane ghosts. A plasma membrane fraction of HeLa S3 cells, consisting of ghosts, is characterized more fully. A simple procedure is described which permits light and electron microscope study of the plasma membrane fraction through the entire depth of the final product pellet and through large areas parallel to the surface. Contamination by nuclei is 0.14%, too little for DNA detection by the diphenylamine reaction. Contamination by rough endoplasmic reticulum and ribosomes is small, a single ghost containing about 3% of the RNA in a single cell. Mitochondria were not encountered. Electron microscopy also shows (a) small vesicles associated with the outer surface of the ghosts, and (b) a filamentous web at the inner face of the ghost membrane. Sodium dodecyl sulfate (SDS)-polyacrylamide gel analysis shows that of the many Coomassie Blue-stained bands two were prominent. One, 43,000 daltons, co-migrated with purified rabbit muscle actin and constituted about 7.5% of the plasma membrane protein. The other major band, 34,000 daltons, was concentrated in the plasma membrane fraction. Two major glycoproteins detected by autoradiography of [14C]fucose-labeled glycoproteins on the gels, had apparent molecular weights of 35,000 daltons and 32,000 daltons. These major bands did not stain with Coomassie Blue. There were many other minor glycoprotein bands in the 200,000- to 80,000-dalton range. Ouabain-sensitive, Na+, K+-adenosine triphosphatase (ATPase) activity of the ghost fraction is purified 9.1 (+/- 2.2) times over the homogenate; recover of the activity is 12.0 (+/- 3.8%) of the homogenate. Enrichment and recovery of fucosylglycoprotein parallel those for ouabain-sensitive Na+, K+-ATPase activity. Fucosyl glycoprotein is recovered more than the enzyme activity in a smooth membrane vesicle fraction probably containing the bulk of plasma membrane not recovered as ghosts.